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R. Woelk, Jana Labrenz, Matthias Juergens, Klaus TI Diversity and abundance of "Pelagibacterales" (SAR11) in the Baltic Sea salinity gradient SO SYSTEMATIC AND APPLIED MICROBIOLOGY DT Article AB The candidate order "Pelagibacterales" (SARI 1) is one of the most abundant bacterial orders in ocean surface waters and, periodically, in freshwater lakes. The presence of several stable phylogenetic lineages comprising "Pelagibacterales" correlates with the physico-chemical parameters in aquatic environments. A previous amplicon sequencing study covering the bacterial community in the salinity gradient of the Baltic Sea suggested that pelagibacteral subclade SAR11-I was replaced by SARI 1-IIIa in the mesohaline region of the Baltic Sea. In this current study, we investigated the cellular abundances of "Pelagibacterales" subclades along the Baltic Sea salinity gradient using catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). The results obtained with a newly designed probe, which exclusively detected SAR11-IIIa, were compared to CARD-FISH abundances of the marine SAR11-I/II subclade and the freshwater lineage SAR11-IIIb (LD12). The results showed that SAR11-IIIa was abundant in oligohaline-mesohaline conditions (salinities 2.7-13.3), with maximal abundances at a salinity of 7 (up to 35% of total Bacteria, quantified with a universal bacterial probe EUB). As expected, SAR11-I/II was abundant (27% of EUB) in the marine parts of the Baltic Sea, whereas counts of the freshwater lineage SAR11-IIIb were below the detection limit at all stations. The shift from SAR11-IIIa to SAR11-I/II was confirmed in the vertical salinity gradient in the deeper basins of the Baltic Sea. These findings were consistent with an overlapping but defined distribution of SARI 1-I/II and SAR11-IIIa in the salinity gradient of the Baltic Sea and suggested the adaptation of SARI 1-IIIa for growth and survival in mesohaline conditions. (C) 2014 Elsevier GmbH. C1 [Herlemann, Daniel P. R.; Woelk, Jana; Labrenz, Matthias; Juergens, Klaus] Leibniz Inst Baltic Sea Res, Dept Biol Oceanog, D-18119 Rostock, Germany. RP Herlemann, DPR (corresponding author), Leibniz Inst Baltic Sea Res, Dept Biol Oceanog, Seestr 15, D-18119 Rostock, Germany. 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Mori, Jiro F. Jessen, Gerdhard L. Nelson, Tara Colenbrander Warren, Lesley A. Banfield, Jillian F. TI Wide Distribution of Phage That Infect Freshwater SAR11 Bacteria SO MSYSTEMS DT Article AB Fonsibacter (LD12 subclade) is among the most abundant bacterioplankton in freshwater ecosystems. These bacteria belong to the order Pelagibacterales (SAR11) and are related to Pelagibacter (marine SAR11), which dominates many marine habitats. Although a few Pelagibacter phage (Pelagiphage) have been described, no phage that infect Fonsibacter have been reported. In this study, we describe two groups of Podoviridae phage that infect Fonsibacter. A complete Fonsibacter genome containing a prophage was reconstructed from metagenomic data. A circularized and complete genome related to the prophage, referred to as uv-Fonsiphage-EPL (lysogenic strategy), shows high similarity to marine Pelagiphage HTVC025P. Additionally, we reconstructed three complete genomes and one draft genome of phage related to marine Pelagiphage HTVC010P and predicted a lytic strategy. The similarity in codon usage and cooccurrence patterns of HTVC010P-related phage and Fonsibacter suggested that these phage infect Fonsibacter. Similar phage were detected in Lake Mendota, Wisconsin, where Fonsibacter is also present. A search of related phage revealed the worldwide distribution of some genotypes in freshwater ecosystems, suggesting their substantial role in shaping indigenous microbial assemblages and influence on biogeochemical cycling. However, the uv-Fonsiphage-EPL and one group of HTVC010P-related phage have a more limited distribution in freshwater ecosystems. Overall, the findings provide insights into the genomic features of phage that infect Fonsibacter and expand understanding of the ecology and evolution of these important bacteria. IMPORTANCE Fonsibacter represents a significant microbial group of freshwater ecosystems. Although the genomic and metabolic features of these bacteria have been well studied, no phage infecting them has been reported. In this study, we reconstructed complete genomes of Fonsibacter and infecting phage and revealed their close relatedness to the phage infecting marine SAR11 members. Also, we illustrated that phage that infect Fonsibacter are widely distributed in freshwater habitats. In summary, the results contribute new insights into the ecology and evolution of Fonsibacter and phage. C1 [Chen, Lin-Xing; Banfield, Jillian F.] Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA. [Zhao, Yanlin] Fujian Agr & Forestry Univ, Fujian Prov Key Lab Agroecol Proc & Safety Monito, Coll Life Sci, Fuzhou, Fujian, Peoples R China. [McMahon, Katherine D.] Univ Wisconsin, Dept Civil & Environm Engn, Madison, WI 53706 USA. [Mori, Jiro F.; Jessen, Gerdhard L.; Nelson, Tara Colenbrander; Warren, Lesley A.] Univ Toronto, Dept Civil & Mineral Engn, Toronto, ON, Canada. [Warren, Lesley A.] McMaster Univ, Sch Geog & Earth Sci, Hamilton, ON, Canada. [Banfield, Jillian F.] Lawrence Berkeley Natl Lab, Earth Sci Div, Berkeley, CA 94720 USA. [Banfield, Jillian F.] Univ Calif Berkeley, Dept Environm Sci Policy & Management, Berkeley, CA 94720 USA. [Banfield, Jillian F.] Chan Zuckerberg Biohub, San Francisco, CA 94158 USA. [Banfield, Jillian F.] Univ Calif Berkeley, Innovat Genom Inst, Berkeley, CA 94720 USA. [Banfield, Jillian F.] Univ Melbourne, Melbourne, Vic, Australia. [McMahon, Katherine D.] Univ Wisconsin, Dept Bacteriol, Madison, WI 53706 USA. [Mori, Jiro F.] Yokohama City Univ, Grad Sch Nanobiosci, Yokohama, Kanagawa, Japan. [Jessen, Gerdhard L.] Univ Austral Chile, Fac Ciencias, Inst Ciencias Marinas & Limnol, Valdivia, Chile. RP Banfield, JF (corresponding author), Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94720 USA.; Banfield, JF (corresponding author), Lawrence Berkeley Natl Lab, Earth Sci Div, Berkeley, CA 94720 USA.; Banfield, JF (corresponding author), Univ Calif Berkeley, Dept Environm Sci Policy & Management, Berkeley, CA 94720 USA.; Banfield, JF (corresponding author), Chan Zuckerberg Biohub, San Francisco, CA 94158 USA.; Banfield, JF (corresponding author), Univ Calif Berkeley, Innovat Genom Inst, Berkeley, CA 94720 USA.; Banfield, JF (corresponding author), Univ Melbourne, Melbourne, Vic, Australia. 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Muehling, Martin Joint, Ian TI A rare SAR11 fosmid clone confirming genetic variability in the 'Candidatus Pelagibacter ubique' genome SO ISME JOURNAL DT Article AB A sequence analysis is described of a fosmid clone from a coastal marine metagenomic library that contains a 16S rRNA gene with high sequence similarity to that of the SAR11 bacterium 'Candidatus Pelagibacter ubique' HTCC1062. The sequence of the fosmid clone was 32 086 bp in length and contained 23 187 bp of the 48-kb hyper-variable region 2 (HVR2) present in the genome of 'Cand. P. ubique'. However, half of the sequences within the HVR2 region of the fosmid clone show little sequence similarity to or have no representative homologues in the genome sequence of 'Cand. P. ubique' HTCC1062. Given their putative functions, the acquisition of these genes suggests that SAR11 could harbour more diverse phenotypes than represented by the 16S rRNA taxonomy. 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James TI The Unique Metabolism of SAR11 Aquatic Bacteria SO JOURNAL OF MICROBIOLOGY DT Review AB The deeply branching clade of abundant, globally distributed aquatic alpha-Proteobacteria known as "SAR11", are adapted to nutrient-poor environments such as the surface waters of the open ocean. Unknown prior to 1990, uncultured until 2002, members of the SAR11 clade can now be cultured in artificial, defined media to densities three orders of magnitude higher than in unamended natural media. Cultivation in natural and defined media has confirmed genomic and metagenomic predictions such as an inability to reduce sulfate to sulfide, a requirement for pyruvate, an ability to oxidize a wide variety of methylated and one-carbon compounds for energy, and an unusual form of conditional glycine auxotrophy. Here we describe the metabolism of the SAR11 type strain Candidatus "Pelagibacter ubique" sir. HTCC1062, as revealed by genome-assisted studies of laboratory cultures. 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Stingl, Ulrich TI Genomic characterization of two novel SAR11 isolates from the Red Sea, including the first strain of the SAR11 Ib clade SO FEMS MICROBIOLOGY ECOLOGY DT Article AB The SAR11 clade (Pelagibacterales) is a diverse group that forms a monophyletic clade within the Alphaproteobacteria, and constitutes up to one third of all prokaryotic cells in the photic zone of most oceans. Pelagibacterales are very abundant in the warm and highly saline surface waters of the Red Sea, raising the question of adaptive traits of SAR11 populations in this water body and warmer oceans through the world. In this study, two pure cultures were successfully obtained from surface waters on the Red Sea: one isolate of subgroup Ia and one of the previously uncultured SAR11 Ib lineage. The novel genomes were very similar to each other and to genomes of isolates of SAR11 subgroup Ia (Ia pan-genome), both in terms of gene content and synteny. Among the genes that were not present in the Ia pan-genome, 108 (RS39, Ia) and 151 genes (RS40, Ib) were strain specific. Detailed analyses showed that only 51 (RS39, Ia) and 55 (RS40, Ib) of these strain-specific genes had not reported before on genome fragments of Pelagibacterales. Further analyses revealed the potential production of phosphonates by some SAR11 members and possible adaptations for oligotrophic life, including pentose sugar utilization and adhesion to marine particulate matter. C1 [Jimenez-Infante, Francy; Ngugi, David Kamanda; Vinu, Manikandan; Stingl, Ulrich] King Abdullah Univ Sci & Technol, Red Sea Res Ctr, Thuwal 239556900, Saudi Arabia. [Blom, Jochen] Justus Liebig Univ, Bioinformat & Syst Biol, D-35392 Giessen, Germany. [Alam, Intikhab; Bajic, Vladimir B.] King Abdullah Univ Sci & Technol, Computat Biosci Res Ctr, Comp Elect & Math Sci & Engn Div, Thuwal 239556900, Saudi Arabia. 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G. Andersson, Siv G. E. TI Independent Genome Reduction and Phylogenetic Reclassification of the Oceanic SAR11 Clade SO MOLECULAR BIOLOGY AND EVOLUTION DT Article AB The SAR11 clade, here represented by Candidatus Pelagibacter ubique, is the most successful group of bacteria in the upper surface waters of the oceans. In contrast to previous studies that have associated the 1.3 Mb genome of Ca. Pelagibacter ubique with the less than 1.5 Mb genomes of the Rickettsiales, our phylogenetic analysis suggests that Ca. Pelagibacter ubique is most closely related to soil and aquatic Alphaproteobacteria with large genomes. This implies that the SAR11 clade and the Rickettsiales have undergone genome reduction independently. A gene flux analysis of 46 representative alphaproteobacterial genomes indicates the loss of more than 800 genes in each of Ca. Pelagibacter ubique and the Rickettsiales. Consistent with their different phylogenetic affiliations, the pattern of gene loss differs with a higher loss of genes for repair and recombination processes in Ca. Pelagibacter ubique as compared with a more extensive loss of genes for biosynthetic functions in the Rickettsiales. Some of the lost genes in Ca. Pelagibacter ubique, such as mutLS, recFN, and ruvABC, are conserved in all other alphaproteobacterial genomes including the small genomes of the Rickettsiales. The mismatch repair genes mutLS are absent from all currently sequenced SAR11 genomes and also underrepresented in the global ocean metagenome data set. We hypothesize that the unique loss of genes involved in repair and recombination processes in Ca. 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Sidhu, Chandni Zhao, Yanlin Eich, Andreas Roessler, Leonard Orellana, Luis H. Fuchs, Bernhard M. TI Globally occurring pelagiphage infections create ribosome-deprived cells SO NATURE COMMUNICATIONS DT Article AB Phages play an essential role in controlling bacterial populations. Those infecting Pelagibacterales (SAR11), the dominant bacteria in surface oceans, have been studied in silico and by cultivation attempts. However, little is known about the quantity of phage-infected cells in the environment. Using fluorescence in situ hybridization techniques, we here show pelagiphage-infected SAR11 cells across multiple global ecosystems and present evidence for tight community control of pelagiphages on the SAR11 hosts in a case study. Up to 19% of SAR11 cells were phage-infected during a phytoplankton bloom, coinciding with a similar to 90% reduction in SAR11 cell abundance within 5 days. Frequently, a fraction of the infected SAR11 cells were devoid of detectable ribosomes, which appear to be a yet undescribed possible stage during pelagiphage infection. We dubbed such cells zombies and propose, among other possible explanations, a mechanism in which ribosomal RNA is used as a resource for the synthesis of new phage genomes. On a global scale, we detected phage-infected SAR11 and zombie cells in the Atlantic, Pacific, and Southern Oceans. Our findings illuminate the important impact of pelagiphages on SAR11 populations and unveil the presence of ribosome-deprived zombie cells as part of the infection cycle. C1 [Bruewer, Jan D.; Sidhu, Chandni; Roessler, Leonard; Orellana, Luis H.; Fuchs, Bernhard M.] Max Planck Inst Marine Microbiol, D-28359 Bremen, Germany. [Zhao, Yanlin] Fujian Agr & Forestry Univ, Coll Juncao Sci & Ecol, Fuzhou, Peoples R China. [Eich, Andreas] PSL Res Univ, EPHE, CNRS, UAR 3278 CRIOBE,UPVD, Moorea, France. 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Freel, Kelle C. Rappe, Michael S. TI Isolation of SAR11 Marine Bacteria from Cryopreserved Seawater SO MSYSTEMS DT Article AB While marine microorganisms are frequently studied in their natural environment, isolated strains are invaluable resources that can be used in controlled experiments to expand upon direct observations from natural systems. Here, we sought a means to enhance culture collections of SAR11 marine bacteria by testing the use of seawater cryopreserved with glycerol as an inoculum. Using a raw seawater sample collected from the tropical Pacific Ocean, a subsample was diluted in seawater growth medium to create 5762-ml dilution cultures containing 5 cells each and incubated for a high-throughput culturing (HTC) experiment, while another portion was cryopreserved in 10% glycerol. After 10 months, a cryopreserved aliquot was thawed and used to create a second cultivation experiment of 480 2-ml cultures containing 5 cells each and 470 cultures containing 105 cells each. The raw seawater cultivation experiment resulted in the successful isolation of 54 monocultures and 29 mixed cultures, while cryopreserved seawater resulted in 59 monocultures and 29 mixed cultures. Combined, the cultures included 51 SAR11 isolates spanning 11 unique 16S rRNA gene amplicon sequence variants (ASVs) from the raw seawater inoculum and 74 SAR11 isolates spanning 13 unique ASVs from cryopreserved seawater. A vast majority (92%) of SAR11 isolates from the two HTC experiments were members of SAR11 subclade la, though subclades lila and Va were also recovered from cryopreserved seawater and subclade lb was recovered from both. The four most abundant SAR11 subclade la ASVs found in the initial seawater environmental sample were isolated by both approaches. IMPORTANCE High-throughput dilution culture has proved to be a successful approach to bring some difficult-to-isolate planktonic microorganisms into culture, including the highly abundant SAR11 lineage of marine bacteria. While the long-term preservation of bacterial isolates by freezing them in the presence of cryoprotectants, such as glycerol, has been shown to be an effective method of storing viable cells over long time periods (i.e., years), to our knowledge it had not previously been tested for its efficacy in preserving raw seawater for later use as an inoculum for high-throughput cultivation experiments. We found that SAR11 and other abundant marine bacteria could be isolated from seawater that was previously cryopreserved for nearly 10 months at a rate of culturability similar to that of the same seawater used fresh, immediately after collection. Our findings (i) expand the potential of high-throughput cultivation experiments to include testing when immediate isolation experiments are impractical, (ii) allow for targeted isolation experiments from specific samples based on analyses such as microbial community structure, and (iii) enable cultivation experiments across a wide range of other conditions that would benefit from having source inocula available over extended periods of time. C1 [Monaghan, Elizabeth A.; Freel, Kelle C.; Rappe, Michael S.] Univ Hawaii Manoa, Hawaii Inst Marine Biol, Sch Ocean & Earth Sci & Technol, Kaneohe, HI 96744 USA. [Monaghan, Elizabeth A.] Univ Hawaii Manoa, Marine Biol Grad Program, Honolulu, HI 96822 USA. RP Rappé, MS (corresponding author), Univ Hawaii Manoa, Hawaii Inst Marine Biol, Sch Ocean & Earth Sci & Technol, Kaneohe, HI 96744 USA. 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Hewson, Ian TI Abundance of Two Pelagibacter ubique Bacteriophage Genotypes along a Latitudinal Transect in the North and South Atlantic Oceans SO FRONTIERS IN MICROBIOLOGY DT Article AB This study characterizes viral and bacterial dynamics along a latitudinal transect in the Atlantic Ocean from approximately 10 N-40 S. Overall viral abundance decreased with depth, on average there were 1.64 +/- 0.71 x 10(7) virus like particles (VLPs) in surface waters, decreasing to an average of 6.50 +/- 2.26 x 10(5) VLPs in Antarctic Bottom Water. This decrease was highly correlated to bacterial abundance. There are six major water masses in the Southern Tropical Atlantic Ocean, and inclusion of water mass, temperature and salinity variables explained a majority of the variation in total viral abundance. Recent discovery of phages infecting bacteria of the SAR11 Glade of Alphaproteobacteria (i.e., pelagiphages) leads to intriguing questions about the roles they play in shaping epipelagic communities. Viral-size fraction DNA from epipelagic water was used to quantify the abundance of two pelagiphages, using pelagiphage-specific quantitative PCR primers and probes along the transect. We found that HTVC010P, a member of a podoviridae sub-family, was most abundant in surface waters. Copy numbers ranged from an average of 1.03 +/- 2.38 x 10(5) copies ml(-1) in surface waters, to 5.79 +/- 2.86 x 10(3) in the deep chlorophyll maximum. HTVC008M, a T4-like myovirus, was present in the deep chlorophyll maximum (5.42 +/- 2.8 x 10(3) copies ml(-1) on average), although it was not as highly abundant as HTVC010P in surface waters (6.05 +/- 3.01 x 10(3) copies ml(-1) on average). 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Pierson, Jason Giovannoni, Stephen J. McIntosh, J. Richard Nicastroa, Daniela TI Three-Dimensional Structure of the Ultraoligotrophic Marine Bacterium "Candidatus Pelagibacter ubique" SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB SAR11 bacteria are small, heterotrophic, marine alphaproteobacteria found throughout the oceans. They thrive at the low nutrient concentrations typical of open ocean conditions, although the adaptations required for life under those conditions are not well understood. To illuminate this issue, we used cryo-electron tomography to study "Candidatus Pelagibacter ubique" strain HTCC1062, a member of the SAR11 clade. Our results revealed its cellular dimensions and details of its intracellular organization. Frozen-hydrated cells, which were preserved in a life-like state, had an average cell volume (enclosed by the outer membrane) of 0.037 +/- 0.011 mu m(3). Strikingly, the periplasmic space occupied similar to 20% to 50% of the total cell volume in log-phase cells and similar to 50% to 70% in stationary-phase cells. The nucleoid occupied the convex side of the crescent-shaped cells and the ribosomes predominantly occupied the concave side, at a relatively high concentration of 10,000 to 12,000 ribosomes/similar to m(3). Outer membrane pore complexes, likely composed of PilQ, were frequently observed in both log-phase and stationary-phase cells. Long filaments, most likely type IV pili, were found on dividing cells. The physical dimensions, intracellular organization, and morphological changes throughout the life cycle of "Ca. Pelagibacter ubique" provide structural insights into the functional adaptions of these oligotrophic ultramicrobacteria to their habitat. IMPORTANCE Bacterioplankton of the SAR11 clade (Pelagibacterales) are of interest because of their global biogeochemical significance and because they appear to have been molded by unusual evolutionary circumstances that favor simplicity and efficiency. They have adapted to an ecosystem in which nutrient concentrations are near the extreme limits at which transport systems can function adequately, and they have evolved streamlined genomes to execute only functions essential for life. However, little is known about the actual size limitations and cellular features of living oligotrophic ultramicrobacteria. In this study, we have used cryo-electron tomography to obtain accurate physical information about the cellular architecture of "Candidatus Pelagibacter ubique," the first cultivated member of the SAR11 clade. These results provide foundational information for answering questions about the cell architecture and functions of these ultrasmall oligotrophic bacteria. 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Bolanos, Luis M. Bell, Ashley G. Michelsen, Michelle L. Allen, Michael J. Temperton, Ben TI Novel pelagiphage isolate Polarivirus skadi is a polar specialist that dominates SAR11-associated bacteriophage communities at high latitudes SO ISME JOURNAL DT Article AB The SAR11 clade are the most abundant members of surface marine bacterioplankton and a critical component of global biogeochemical cycles. Similarly, pelagiphages that infect SAR11 are ubiquitous and highly abundant in the oceans. Pelagiphages are predicted to shape SAR11 community structures and increase carbon turnover throughout the oceans. Yet, ecological drivers of host and niche specificity of pelagiphage populations are poorly understood. Here we report the global distribution of a novel pelagiphage called "Polarivirus skadi", which is the sole representative of a novel genus. P. skadi was isolated from the Western English Channel using a cold-water ecotype of SAR11 as bait. 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Weaver, Alec J. Davis, Edward W. Giovannoni, Stephen J. Halsey, Kimberly H. TI Metabolism of key atmospheric volatile organic compounds by the marine heterotrophic bacterium Pelagibacter HTCC1062 (SAR11) SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Plants and phytoplankton are natural sources of the volatile organic compounds (VOCs) acetone and isoprene, which are reactive and can alter atmospheric chemistry. In earlier research we reported that, when co-cultured with a diatom, the marine bacterium Pelagibacter (strain HTCC1062; 'SAR11 clade') reduced the concentration of compounds tentatively identified as acetone and isoprene. In this study, experiments with Pelagibacter monocultures confirmed that these cells are capable of metabolizing acetone and isoprene at rates similar to bacterial communities in seawater and high enough to consume substantial fractions of the total marine acetone and isoprene budgets if extrapolated to global SAR11 populations. Homologues of an acetone/cyclohexanone monooxygenase were identified in the HTCC1062 genome and in the genomes of a wide variety of other abundant marine taxa, and were expressed at substantial levels (c. 10(-4) of transcripts) across TARA oceans metatranscriptomes from ocean surface samples. The HTCC1062 genome lacks the canonical isoprene degradation pathway, suggesting an unknown alternative biochemical pathway is used by these cells for isoprene uptake. Fosmidomycin, an inhibitor of bacterial isoprenoid biosynthesis, blocked HTCC1062 growth, but the cells were rescued when isoprene was added to the culture, indicating SAR11 cells may be capable of synthesizing isoprenoid compounds from exogenous isoprene. C1 [Moore, Eric R.; Weaver, Alec J.; Davis, Edward W.; Giovannoni, Stephen J.; Halsey, Kimberly H.] Oregon State Univ, Dept Microbiol, 226 Nash Hall, Corvallis, OR 97331 USA. [Halsey, Kimberly H.] Los Alamos Natl Lab, Biosci Div, Los Alamos, NM 87545 USA. 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Freel, Kelle C. Monaghan, Elizabeth A. Sullivan, Clarisse E. S. Ramfelt, Oscar Rii, Yoshimi M. Rappe, Michael S. TI Spatial and temporal dynamics of SAR11 marine bacteria across a nearshore to offshore transect in the tropical Pacific Ocean SO PEERJ DT Article AB Surveys of microbial communities across transitions coupled with contextual measures of the environment provide a useful approach to dissect the factors determining distributions of microorganisms across ecological niches. Here, monthly time-series samples of surface seawater along a transect spanning the nearshore coastal environment within Kane.ohe Bay on the island of O.ahu, Hawai.i, and the adjacent offshore environment were collected to investigate the diversity and abundance of SAR11 marine bacteria (order Pelagibacterales) over a 2-year time period. Using 16S ribosomal RNA gene amplicon sequencing, the spatiotemporal distributions of major SAR11 subclades and exact amplicon sequence variants (ASVs) were evaluated. 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Meier, David A. O. Lanclos, V. Celeste Asrari, Hasti Coates, John D. Thrash, J. Cameron TI Polyhydroxybutyrate production by freshwater SAR11 (LD12) SO ISME JOURNAL DT Article AB SAR11 bacteria (order Pelagibacterales) are oligotrophs and often the most abundant bacterioplankton in aquatic environments. A subset of sequenced SAR11 genomes, predominantly in the brackish and freshwater SAR11 subclades, contain homologs of pha genes, which in other organisms confer the ability to store carbon and energy via polyhydroxyalkanoate (PHA) polymers. Here, we investigated the relevance of PHA production to SAR11 biology. Phylogenetics showed that Pha proteins occurred on a long branch and provided evidence for origin at the common ancestor of the brackish IIIa and freshwater LD12 subclades, followed by horizontal transfer within SAR11. Using the LD12 representative "Candidatus Fonsibacter ubiquis" strain LSUCC0530, we found that many LSUCC0530 cells contained a single Nile red-staining granule, confirmed that the cells produced polyhydroxybutyrate, a common form of PHA, and estimated the total polyhydroxybutyrate content in the cells. We heterologously expressed the LSUCC0530 phaCAB locus in Escherichia coli, finding it to be functional and the likely origin of the polyhydroxybutyrate. We also determined that, irrespective of changes to carbon, nitrogen, and phosphorus concentrations, a similar fraction of LSUCC0530 cells generated polyhydroxybutyrate granules and expression of the phaCAB locus remained constant. We suggest that polyhydroxybutyrate synthesis in LSUCC0530 may be constitutively active due to the slow growth dynamics and minimal regulation that characterize SAR11 bacteria. This work characterizes polymer storage in SAR11, providing new insights into the likely fitness advantage for cells harboring this metabolism. C1 [Bennett, Brittany D.; Lanclos, V. Celeste; Asrari, Hasti; Thrash, J. Cameron] Univ Southern Calif, Dept Biol Sci, 3616 Trousdale Pkwy, AHF 107, Los Angeles, CA 90089 USA. [Meier, David A. O.; Coates, John D.] Univ Calif Berkeley, Dept Plant & Microbial Biol, Berkeley, CA 94720 USA. RP Thrash, JC (corresponding author), Univ Southern Calif, Dept Biol Sci, 3616 Trousdale Pkwy, AHF 107, Los Angeles, CA 90089 USA. 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Nicora, Carrie D. Carini, Paul Lipton, Mary S. Norbeck, Angela D. Smith, Richard D. Giovannoni, Stephen J. TI Proteome Remodeling in Response to Sulfur Limitation in "Candidatus Pelagibacter ubique" SO MSYSTEMS DT Article AB The alphaproteobacterium "Candidatus Pelagibacter ubique" strain HTCC1062 and most other members of the SAR11 clade lack genes for assimilatory sulfate reduction, making them dependent on organosulfur compounds that occur naturally in seawater. To investigate how these cells adapt to sulfur limitation, batch cultures were grown in defined medium containing either limiting or nonlimiting amounts of dimethylsulfoniopropionate (DMSP) as the sole sulfur source. Protein and mRNA expression were measured before, during, and after the transition from exponential growth to stationary phase. Two distinct responses were observed, one as DMSP became exhausted and another as the cells acclimated to a sulfur-limited environment. The first response was characterized by increased transcription and translation of all "Ca. Pelagibacter ubique" genes downstream from the previously confirmed S-adenosyl methionine (SAM) riboswitches bhmT, mmuM, and metY. The proteins encoded by these genes were up to 33 times more abundant as DMSP became limiting. Their predicted function is to shunt all available sulfur to methionine. The secondary response, observed during sulfur-limited stationary phase, was a 6- to 10-fold increase in the transcription of the heme c shuttle-encoding gene ccmC and two small genes of unknown function (SAR11_1163 and SAR11_1164). This bacterium's strategy for coping with sulfur stress appears to be intracellular redistribution to support methionine biosynthesis rather than increasing organosulfur import. Many of the genes and SAM riboswitches involved in this response are located in a hypervariable genome region (HVR). One of these HVR genes, ordL, is located downstream from a conserved motif that evidence suggests is a novel riboswitch. IMPORTANCE "Ca. Pelagibacter ubique" is a key driver of marine biogeochemistry cycles and a model for understanding how minimal genomes evolved in free-living anucleate organisms. This study explores the unusual sulfur acquisition strategy that has evolved in these cells, which lack assimilatory sulfate reduction and instead rely on reduced sulfur compounds found in oxic marine environments to meet their cellular quotas. Our findings demonstrate that the sulfur acquisition systems are constitutively expressed but the enzymatic steps leading to the essential sulfur-containing amino acid methionine are regulated by a unique array of riboswitches and genes, many of which are encoded in a rapidly evolving genome region. 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Lanclos, V. Celeste Kojima, Conner Y. Cheng, Chuankai Henson, Michael W. Schon, Max Emil Ettema, Thijs J. G. Faircloth, Brant C. Thrash, J. Cameron TI The AEGEAN-169 clade of bacterioplankton is synonymous with SAR11 subclade V (HIMB59) and metabolically distinct SO MSYSTEMS DT Article AB Bacterioplankton of the SAR11 clade are the most abundant marine microorganisms and consist of numerous subclades spanning order-level divergence (Pelagibacterales). The assignment of the earliest diverging subclade V (a.k.a. HIMB59) to the Pelagibacterales is highly controversial, with multiple recent phylogenetic studies placing them completely separate from SAR11. Other than through phylogenomics, subclade V has not received detailed examination due to limited genomes from this group. Here, we assessed the ecogenomic characteristics of subclade V to better understand the role of this group in comparison to the Pelagibacterales. We used a new isolate genome, recently released single-amplified genomes and metagenome-assembled genomes, and previously established SAR11 genomes to perform a comprehensive comparative genomics analysis. We paired this analysis with the recruitment of metagenomes spanning the open ocean, coastal, and brackish systems. Phylogenomics, average amino acid identity, and 16S rRNA gene phylogeny indicate that SAR11 subclade V is synonymous with the ubiquitous AEGEAN-169 clade and support the contention that this group represents a taxonomic family. AEGEAN-169 shared many bulk genome qualities with SAR11, such as streamlining and low GC content, but genomes were generally larger. AEGEAN-169 had overlapping distributions with SAR11 but was metabolically distinct from SAR11 in its potential to transport and utilize a broader range of sugars as well as in the transport of trace metals and thiamin. Thus, regardless of the ultimate phylogenetic placement of AEGEAN-169, these organisms have distinct metabolic capacities that likely allow them to differentiate their niche from canonical SAR11 taxa. IMPORTANCE One goal of marine microbiologists is to uncover the roles various microorganisms are playing in biogeochemical cycles. Success in this endeavor relies on differentiating groups of microbes and circumscribing their relationships. An early-diverging group (subclade V) of the most abundant bacterioplankton, SAR11, has recently been proposed as a separate lineage that does not share a most recent common ancestor. But beyond phylogenetics, little has been done to evaluate how these organisms compare with SAR11. Our work leverages dozens of new genomes to demonstrate the similarities and differences between subclade V and SAR11. In our analysis, we also establish that subclade V is synonymous with a group of bacteria established from 16S rRNA gene sequences, AEGEAN-169. Subclade V/AEGEAN-169 has clear metabolic distinctions from SAR11 and their shared traits point to remarkable convergent evolution if they do not share a most recent common ancestor. C1 [Getz, Eric W.; Lanclos, V. Celeste; Kojima, Conner Y.; Cheng, Chuankai; Thrash, J. Cameron] Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA. [Henson, Michael W.] Univ Chicago, Dept Geophys Sci, Chicago, IL USA. [Schon, Max Emil] Uppsala Univ, Sci Life Lab, Dept Cell & Mol Biol, Uppsala, Sweden. [Ettema, Thijs J. G.] Wageningen Univ & Res, Lab Microbiol, Wageningen, Netherlands. [Faircloth, Brant C.] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA USA. [Faircloth, Brant C.] Louisiana State Univ, Museum Nat Sci, Baton Rouge, LA USA. RP Thrash, JC (corresponding author), Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA. 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Barrell, Gregory E. Suffridge, Christopher Morre, Jeff Gable, Kevin P. Graff, Jason R. VerWey, Brian J. Hellweger, Ferdi L. Giovannoni, Stephen J. TI SAR11 Cells Rely on Enzyme Multifunctionality To Metabolize a Range of Polyamine Compounds SO MBIO DT Article AB In the ocean surface layer and cell culture, the polyamine transport protein PotD of SAR11 bacteria is often one of the most abundant proteins detected. Polyamines are organic cations at seawater pH produced by all living organisms and are thought to be an important component of dissolved organic matter (DOM) produced in planktonic ecosystems. We hypothesized that SAR11 cells uptake and metabolize multiple poly amines and use them as sources of carbon and nitrogen. Metabolic footprinting and fin-gerprinting were used to measure the uptake of five polyamine compounds (putrescine, cadaverine, agmatine, norspermidine, and spermidine) in two SAR11 strains that represent the majority of SAR11 cells in the surface ocean environment, "Candidatus Pelagibacter" strain HTCC7211 and "Candidatus Pelagibacter ubique" strain HTCC1062. Both strains took up all five polyamines and concentrated them to micromolar or milli molar intracellular concentrations. Both strains could use most of the polyamines to meet their nitrogen requirements, but polyamines did not fully substitute for their requirements of glycine (or related compounds) or pyruvate (or related compounds). Our data suggest that potABCD transports all five polyamines and that spermidine synthase, speE, is reversible, catalyzing the breakdown of spermidine and norspermidine, in addition to its usual biosynthetic role. These findings provide support for the hypothesis that enzyme multifunctionality enables streamlined cells in planktonic ecosystems to increase the range of DOM compounds they metabolize. IMPORTANCE Genome streamlining in SAR11 bacterioplankton has resulted in a small repertoire of genes, yet paradoxically, they consume a substantial fraction of primary production in the oceans. Enzyme multifunctionality, referring to enzymes that are adapted to have broader substrate and catalytic range than canonically defined, is hypothesized to be an adaptation that increases the range of organic compounds metabolized by cells in environments where selection favors genome minimization. We provide experimental support for this hypothesis by demonstrating that SAR11 cells take up and metabolize multiple polyamine compounds and propose that a small set of multifunctional enzymes catalyze this metabolism. 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Due to the lack of a universal conserved gene among all viruses, metagenomic fragment recruitment is commonly used to estimate the relative viral abundance. However, the paucity of absolute virus abundance data hinders our ability to fully understand how viruses drive global microbial populations. The cosmopolitan marine Pelagibacter ubique is host for the highly widespread HTVC010P pelagiphage isolate and the extremely abundant uncultured virus vSAG 37-F6 recently discovered by single-virus genomics. Here we applied droplet digital PCR (ddPCR) to calculate the absolute abundance of these pelagiphage genotypes in the Mediterranean Sea and the Gulf of Maine. Abundances were between 360 and 8,510 virus mL-1 and 1,270-14,400 virus mL-1 for vSAG 37-F6 and HTVC010P, respectively. Illumina PCR-amplicon sequencing corroborated the absence of ddPCR non-specific amplifications for vSAG 37-F6, but showed an overestimation of 6% for HTVC010P from off-targets, genetically unrelated viruses. 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Lopez-Perez, Mario Molina-Pardines, Carmen Rodriguez-Valera, Francisco TI Large diversity in the O-chain biosynthetic cluster within populations of Pelagibacterales SO MBIO DT Article AB Genomic diversity in prokaryotic species is largely due to the existence of extensive pangenomes, allowing different gene complements to be drawn depend ing on the strain. Here, we have studied the diversity of the O-chain polysaccharide biosynthesis cluster (OBC) in marine bacteria of the Pelagibacterales order as a proxy to measure such genetic diversity in a single population. The study of single-ampli fied genomes (SAGs) from the whole order found a pattern similar to that of other well-studied microbes, such as the Enterobacteriales or Alteromonas, where distinct OBCs represent strains containing different gene pools. We found that most of the OBC sharing happened among individuals of the same clonal frame (>99% average nucleotide identity). Moreover, given the parsimonious way this cluster changes, the diversity of the OBCs can be extrapolated to the size of the population's pangenome. This assumes that different OBCs correspond to lineages containing unique flexible gene pools, as seen in the aforementioned microbes. Through long-read metagenomics, we could detect 380 different OBCs at a single Mediterranean sampling site. Within a single population (single species and sample) of the endemic Ia.3/VII (gMED) genomospecies, we identified 158 OBCs, of which 130 were unique. These findings suggest that the gene pool within a single population might be substantial (several thousands). While this figure is large, it aligns with the complexity of the dissolved organic matter that these organisms can potentially degrade. C1 [Haro-Moreno, Jose M.; Lopez-Perez, Mario; Molina-Pardines, Carmen; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Div Microbiol, Evolutionary Genom Grp, Alicante, Spain. 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Cameron Schwalbach, Michael S. Vergin, Kevin L. Landry, Zachary C. Ellisman, Mark Deerinck, Tom Sullivan, Matthew B. Giovannoni, Stephen J. TI Abundant SAR11 viruses in the ocean SO NATURE DT Article AB Several reports proposed that the extraordinary dominance of the SAR11 bacterial clade in ocean ecosystems could be a consequence of unusual mechanisms of resistance to bacteriophage infection, including 'cryptic escape' through reduced cell size(1) and/or K-strategist defence specialism(2). Alternatively, the evolution of high surface-to-volume ratios coupled with minimal genomes containing high-affinity transporters enables unusually efficient metabolism for oxidizing dissolved organic matter in the world's oceans that could support vast population sizes despite phage susceptibility. These ideas are important for understanding plankton ecology because they emphasize the potentially important role of top-down mechanisms in predation, thus determining the size of SAR11 populations and their concomitant role in biogeochemical cycling. Here we report the isolation of diverse SAR11 viruses belonging to two virus families in culture, for which we propose the name 'pelagiphage', after their host. Notably; the pelagiphage genomes were highly represented in marine viral metagenomes, demonstrating their importance in nature. One of the new phages, HTVC010P, represents a new podovirus subfamily more abundant than any seen previously, in all data sets tested, and may represent one of the most abundant virus subfamilies in the biosphere. This discovery disproves the theory that SAR11 cells are immune to viral predation and is consistent with the interpretation that the success of this highly abundant microbial clade is the result of successfully evolved adaptation to resource competition. C1 [Zhao, Yanlin; Temperton, Ben; Thrash, J. Cameron; Vergin, Kevin L.; Landry, Zachary C.; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Schwalbach, Michael S.] Univ Wisconsin, Great Lakes Bioenergy Res Ctr, Madison, WI 53706 USA. [Ellisman, Mark; Deerinck, Tom] Univ Calif San Diego, Natl Ctr Microscopy & Imaging Res, San Diego, CA 92093 USA. [Sullivan, Matthew B.] Univ Arizona, Dept Ecol & Evolutionary Biol, Tucson, AZ 85721 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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Here, we report that Pelagibacter HTCC1062 produces the gas methanethiol, and that a second DMSP catabolic pathway, mediated by a cupin-like DMSP lyase, DddK, simultaneously shunts as much as 59% of DMSP uptake to dimethyl sulfide production. We propose a model in which the allocation of DMSP between these pathways is kinetically controlled to release increasing amounts of dimethyl sulfide as the supply of DMSP exceeds cellular sulfur demands for biosynthesis. C1 [Sun, Jing; Davie-Martin, Cleo L.; Halsey, Kimberly H.; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Todd, Jonathan D.; Fowler, Emily K.; Johnston, Andrew W. B.] Univ East Anglia, Sch Biol Sci, Norwich Res Pk, Norwich NR4 7TJ, Norfolk, England. [Thrash, J. Cameron] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA. [Qian, Yanping; Qian, Michael C.] Oregon State Univ, Dept Food Sci, Corvallis, OR 97331 USA. [Temperton, Ben] Univ Exeter, Dept Biosci, Exeter EX4 4QD, Devon, England. [Guo, Jiazhen] Qingdao Aquarium, Qingdao 266003, Shandong, Peoples R China. [Aldrich, Joshua T.; Nicora, Carrie D.; Lipton, Mary S.; Smith, Richard D.; Payne, Samuel H.] Pacific Northwest Natl Lab, Environm Mol Sci Lab, Richland, WA 99352 USA. [De Leenheer, Patrick] Oregon State Univ, Dept Math, Corvallis, OR 97331 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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It can be metabolized by marine bacteria into volatile methylated amines, the second largest source of nitrogen after N-2 gas in the oceans. The SAR11 bacteria are the most abundant oligotrophic plankton in the oceans, which represents approximately 30% of the bacterial cells in marine surface waters. Genomic analysis suggested that most SAR11 bacteria possess an ATP-binding cassette transporter TmoXWV that may be responsible for importing TMAO. However, it was still unclear whether SAR11 bacteria can utilize TMAO as the sole nitrogen source and how they import TMAO. Here, our results showed that Pelagibacter strain HTCC1062, a SAR11 bacterium, can grow with TMAO as the sole nitrogen source. TmoXWV from strain HTCC1062 (TmoXWV(1062)) was verified to be a functional TMAO importer. Furthermore, TmoX(1062), the periplasmic substrate binding protein of TmoXWV(1062), was shown to have high binding affinities toward TMAO at 4 degrees C (K-d = 920 nM), 10 degrees C (K-d = 500 nM) and 25 degrees C (K-d = 520 nM). The high TMAO binding affinity and strong temperature adaptability of TmoX(1062) reveal a possible oligotrophic niche adaptation strategy of strain HTCC1062, which may help it gain a competitive advantage over other bacteria. Structure comparison and mutational analysis indicated that the TMAO binding mechanism of TmoX(1062) may have differences from the previously reported mechanism of TmoX of Ruegeria pomeroyi DSS-3. This study provides new insight into TMAO utilization by the widespread SAR11 bacteria. C1 [Gao, Chao; He, Xiao-Yan; Wang, Ning; Zhang, Xi-Ying; Wang, Peng; Chen, Xiu-Lan; Zhang, Yu-Zhong] Shandong Univ, Marine Biotechnol Res Ctr, State Key Lab Microbial Technol, Qingdao, Peoples R China. [Gao, Chao; Wang, Peng; Zhang, Yu-Zhong; Li, Chun-Yang] Ocean Univ China, Coll Marine Life Sci, Frontiers Sci Ctr Deep Ocean Multispheres & Earth, Qingdao, Peoples R China. [Gao, Chao; Chen, Xiu-Lan; Zhang, Yu-Zhong; Li, Chun-Yang] Pilot Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China. [Zhang, Nan] Qilu Univ Technol, Sch Bioengn, Jinan, Peoples R China. [Ding, Jun-Mei] Yunnan Normal Univ, Engn Res Ctr Sustainable Dev & Utilizat Biomass E, Minist Educ, Kunming, Peoples R China. RP Li, CY (corresponding author), Ocean Univ China, Coll Marine Life Sci, Frontiers Sci Ctr Deep Ocean Multispheres & Earth, Qingdao, Peoples R China.; Li, CY (corresponding author), Pilot Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China.; Ding, JM (corresponding author), Yunnan Normal Univ, Engn Res Ctr Sustainable Dev & Utilizat Biomass E, Minist Educ, Kunming, Peoples R China. 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Hernandes Coutinho, Felipe Martinez-Garcia, Manuel Rodriguez-Valera, Francisco TI The Evolutionary Success of the Marine Bacterium SAR11 Analyzed through a Metagenomic Perspective SO MSYSTEMS DT Article AB The SAR11 clade of Alphaproteobacteria is the most abundant group of planktonic cells in the near-surface epipelagic waters of the ocean, but the mechanisms underlying its exceptional success have not been fully elucidated. Here, we applied a metagenomic approach to explore microdiversity patterns by measuring the accumulation of synonymous and nonsynonymous mutations as well as homologous recombination in populations of SAR11 from different aquatic habitats (marine epipelagic, bathypelagic, and surface freshwater). The patterns of mutation accumulation and recombination were compared to those of other groups of representative marine microbes with multiple ecological strategies that share the same marine habitat, namely, Cyanobacteria (Prochlorococcus and Synechococcus), Archaea ("Candidatus Nitrosopelagicus" and Marine Group II Thalassoarchaea), and some heterotrophic marine bacteria (Alteromonas and Erythrobacter). SAR11 populations showed widespread recombination among distantly related members, preventing divergence leading to a genetically stable population. Moreover, their high intrapopulation sequence diversity with an enrichment in synonymous replacements supports the idea of a very ancient divergence and the coexistence of multiple different clones. However, other microbes analyzed seem to follow different evolutionary dynamics where processes of diversification driven by geographic and ecological instability produce a higher number of nonsynonymous replacements and lower intrapopulation sequence diversity. Together, these data shed light on some of the evolutionary and ecological processes that lead to the large genomic diversity in SAR11. Furthermore, this approach can be applied to other similar microbes that are difficult to culture in the laboratory, but abundant in nature, to investigate the underlying dynamics of their genomic evolution. IMPORTANCE As the most abundant bacteria in oceans, the Pelagibacterales order (here SAR11) plays an important role in the global carbon cycle, but the study of the evolutionary forces driving its evolution has lagged considerably due to the inherent difficulty of obtaining pure cultures. Multiple evolutionary models have been proposed to explain the diversification of distinct lineages within a population; however, the identification of many of these patterns in natural populations remains mostly enigmatic. We have used a metagenomic approach to explore microdiversity patterns in their natural habitats. Comparison with a collection of bacterial and archaeal groups from the same environments shows that SAR11 populations have a different evolutionary regime, where multiple genotypes coexist within the same population and remain stable over time. Widespread homologous recombination could be one of the main driving factors of this homogenization. C1 [Lopez-Perez, Mario; Haro-Moreno, Jose M.; Hernandes Coutinho, Felipe; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Div Microbiol, Evolutionary Genom Grp, Alicante, Spain. [Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. 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Strain HTCC7211 SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB In the marine environment, phosphorus availability significantly affects the lipid composition in many cosmopolitan marine heterotrophic bacteria, including members of the SAR11 Glade and the Roseobacter Glade. Under phosphorus stress conditions, nonphosphorus sugar-containing glycoglycerolipids are substitutes for phospholipids in these bacteria. Although these glycoglycerolipids play an important role as surrogates for phospholipids under phosphate deprivation, glycoglycerolipid synthases in marine microbes are poorly studied. In the present study, we biochemically characterized a glycolipid glycosyltransferase (GT(cp)) from the marine bacterium "Candidatus Pelagibacter sp." strain HTCC7211, a member of the SAR11 Glade. Our results showed that GT(cp) is able to act as a multifunctional enzyme by synthesizing different glycoglycerolipids with UDP-glucose, UDP-galactose, or UDP-glucuronic acid as sugar donors and diacylglycerol (DAG) as the acceptor. Analyses of enzyme kinetic parameters demonstrated that Mg2+ notably changes the enzyme's affinity for UDP-glucose, which improves its catalytic efficiency. Homology modeling and mutational analyses revealed binding sites for the sugar donor and the diacylglycerol lipid acceptor, which provided insights into the retaining mechanism of GT(cp) with its GT-B fold. A phylogenetic analysis showed that GT(cp) and its homologs form a group in the GT4 glycosyltransferase family. These results not only provide new insights into the glycoglycerolipid synthesis mechanism in lipid remodeling but also describe an efficient enzymatic tool for the future synthesis of bioactive molecules. IMPORTANCE The bilayer formed by membrane lipids serves as the containment unit for living microbial cells. In the marine environment, it has been firmly established that phytoplankton and heterotrophic bacteria can replace phospholipids with nonphosphorus sugar-containing glycoglycerolipids in response to phosphorus limitation. However, little is known about how these glycoglycerolipids are synthesized. Here, we determined the biochemical characteristics of a glycolipid glycosyltransferase (GT(cp)) from the marine bacterium "Candidatus Pelagibacter sp." strain HTCC7211. GT(cp) and its homologs form a group in the GT4 glycosyltransferase family and can synthesize neutral glycolipids (monoglucosyl-1,2-diacyl-sn-glycerol [MGlc-DAG] and monogalactosyl [MGaI]-DAG) and monoglucuronic acid diacylglycerol (MGlcA-DAG). We also uncovered the key residues for DAG binding through molecular docking, site-direct mutagenesis, and subsequent enzyme activity assays. Our data provide new insights into the glycoglycerolipid synthesis mechanism in lipid remodeling. C1 [Wei, Tao; Zhao, Caimeng; Wu, Nan; Huang, Shen; Zhao, Yuezhe; Yang, Pengfei; Mao, Duobin] Zhengzhou Univ Light Ind, Sch Food & Biol Engn, Zhengzhou, Peoples R China. [Quareshy, Mussa; Chen, Yin] Univ Warwick, Sch Life Sci, Coventry, W Midlands, England. RP Wei, T (corresponding author), Zhengzhou Univ Light Ind, Sch Food & Biol Engn, Zhengzhou, Peoples R China.; Chen, Y (corresponding author), Univ Warwick, Sch Life Sci, Coventry, W Midlands, England. 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Thrash, J. Cameron Nicora, Carrie D. Lipton, Mary S. Burnum-Johnson, Kristin E. Carini, Paul Smith, Richard D. Giovannoni, Stephen J. TI Proteomic and Transcriptomic Analyses of "Candidatus Pelagibacter ubique" Describe the First PII-Independent Response to Nitrogen Limitation in a Free-Living Alphaproteobacterium SO MBIO DT Article AB Nitrogen is one of the major nutrients limiting microbial productivity in the ocean, and as a result, most marine microorganisms have evolved systems for responding to nitrogen stress. The highly abundant alphaproteobacterium "Candidatus Pelagibacter ubique," a cultured member of the order Pelagibacterales (SAR11), lacks the canonical GlnB, GlnD, GlnK, and NtrB/NtrC genes for regulating nitrogen assimilation, raising questions about how these organisms respond to nitrogen limitation. A survey of 266 Alphaproteobacteria genomes found these five regulatory genes nearly universally conserved, absent only in intracellular parasites and members of the order Pelagibacterales, including "Ca. Pelagibacter ubique." Global differences in mRNA and protein expression between nitrogen-limited and nitrogen-replete cultures were measured to identify nitrogen stress responses in "Ca. Pelagibacter ubique" strain HTCC1062. Transporters for ammonium (AmtB), taurine (TauA), amino acids (YhdW), and opines (OccT) were all elevated in nitrogen-limited cells, indicating that they devote increased resources to the assimilation of nitrogenous organic compounds. Enzymes for assimilating amine into glutamine (GlnA), glutamate (GltBD), and glycine (AspC) were similarly upregulated. Differential regulation of the transcriptional regulator NtrX in the two-component signaling system NtrY/NtrX was also observed, implicating it in control of the nitrogen starvation response. Comparisons of the transcriptome and proteome supported previous observations of uncoupling between transcription and translation in nutrient-deprived "Ca. Pelagibacter ubique" cells. Overall, these data reveal a streamlined, P-II-independent response to nitrogen stress in "Ca. Pelagibacter ubique," and likely other Pelagibacterales, and show that they respond to nitrogen stress by allocating more resources to the assimilation of nitrogen-rich organic compounds. IMPORTANCE Pelagibacterales are extraordinarily abundant and play a pivotal role in marine geochemical cycles, as one of the major recyclers of labile dissolved organic matter. They are also models for understanding how streamlining selection can reshape chemoheterotroph metabolism. Streamlining and its broad importance to environmental microbiology are emerging slowly from studies that reveal the complete genomes of uncultured organisms. Here, we report another remarkable example of streamlined metabolism in Pelagibacterales, this time in systems that control nitrogen assimilation. Pelagibacterales are major contributors to metatranscriptomes and metaproteomes from ocean systems, where patterns of gene expression are used to gain insight into ocean conditions and geochemical cycles. The data presented here supply background that is essential to interpreting data from field studies. C1 [Smith, Daniel P.; Thrash, J. Cameron; Carini, Paul; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Nicora, Carrie D.; Lipton, Mary S.; Burnum-Johnson, Kristin E.; Smith, Richard D.] Pacific NW Natl Lab, Biol & Computat Sci Div, Richland, WA 99352 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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Manrique, Julieta M. TI High-throughput metabarcoding of SAR11 assemblages from the southwest Atlantic shelf and arid Patagonia: richness and associated rank abundance distributions SO QUANTITATIVE BIOLOGY DT Article AB Background: Massively parallel sequencing of environmental DNA allows microbiological studies to be performed in greater detail than was possible with first-generation sequencing. For example, it facilitates the use of approaches hitherto largely applied to flora and fauna, such as rank abundance distribution (RAD) analyses. Methods: Here, we set out to advance the knowledge on Ca. Pelagibacterales (SAR11) communities from southern South America using environmental sequences from the open ocean in the Argentine sea, the uncharted Engano Bay, as well as a river and an oligohaline shallow lake from the Patagonian Steppe ecoregion. The structures of the SAR11 assemblages present in these ecosystems were dissected by direct and rarefaction-based estimates of species richness, and evaluations of the corresponding abundance distributions (ADs), which was addressed by RAD analyses.Results: Microbial community composition analyses revealed that the studied SAR11 assemblages coexist with 27 bacterial phyla. SAR11 richness was in general very high, but ADs turned out to be highly uneven. The results were compatible with prior knowledge, and similar to that derived from point estimates of diversity. However, our comprehensive dissection allowed for more detailed quantitative comparisons to be made between the environments surveyed, and revealed differences regarding both richness and the underlying ADs.Conclusions: Despite SAR11 assemblages being extremely rich, their ADs are very uneven. Richness and ADs can vary, not only between fresh and salt water, but also between oceanic and coastal marine environments. 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Measurements of cellular ATP content and the oxidation of C-14-labeled compounds to (CO2)-C-14 indicated that methanol, formaldehyde, methylamine, and methyl groups from glycine betaine (GBT), trimethylamine (TMA), trimethylamine N-oxide (TMAO), and dimethylsulfoniopropionate (DMSP) were oxidized by axenic cultures of the SAR11 strain Ca. P. ubique HTCC1062. Analyses of metagenomic data showed that genes for C1 metabolism occur at a high frequency in natural SAR11 populations. In short term incubations, natural communities of Sargasso Sea microbial plankton expressed a potential for the oxidation of C-14-labeled formate, formaldehyde, methanol and TMAO that was similar to cultured SAR11 cells and, like cultured SAR11 cells, incorporated a much larger percentage of pyruvate and glucose (27-35%) than of C1 compounds (2-6%) into biomass. Collectively, these genomic, cellular and environmental data show a surprising capacity for demethylation and C1 oxidation in SAR11 cultures and in natural microbial communities dominated by SAR11, and support the conclusion that C1 oxidation might be a significant conduit by which dissolved organic carbon is recycled to CO2 in the upper ocean. C1 [Sun, Jing; Steindler, Laura; Thrash, J. Cameron; Smith, Daniel P.; Carter, Amy E.; Landry, Zachary C.; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Halsey, Kimberly H.] Oregon State Univ, Dept Bot & Plant Pathol, Corvallis, OR 97331 USA. RP Sun, J (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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Campbell, Emily O. Giovannoni, Stephen J. TI Methane production by phosphate-starved SAR11 chemoheterotrophic marine bacteria SO NATURE COMMUNICATIONS DT Article AB The oxygenated surface waters of the world's oceans are supersaturated with methane relative to the atmosphere, a phenomenon termed the 'marine methane paradox'. The production of methylphosphonic acid (MPn) by marine archaea related to Nitrosopumilus maritimus and subsequent decomposition of MPn by phosphate-starved bacterioplankton may partially explain the excess methane in surface waters. Here we show that Pelagibacterales sp. strain HTCC7211, an isolate of the SAR11 clade of marine alpha-proteobacteria, produces methane from MPn, stoichiometric to phosphorus consumption, when starved for phosphate. Gene transcripts encoding phosphonate transport and hydrolysis proteins are upregulated under phosphate limitation, suggesting a genetic basis for the methanogenic phenotype. Strain HTCC7211 can also use 2-aminoethylphosphonate and assorted phosphate esters for phosphorus nutrition. Despite strain-specific differences in phosphorus utilization, these findings identify Pelagibacterales bacteria as a source of biogenic methane and further implicate phosphate starvation of chemoheterotrophic bacteria in the long-observed methane supersaturation in oxygenated waters. C1 [Carini, Paul; Campbell, Emily O.; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [White, Angelicque E.] Oregon State Univ, Coll Earth, Ocean & Atmospher Sci, Corvallis, OR 97331 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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Terminal restriction fragment length polymorphism (T-RFLP) data generated from a decade of samples collected at BATS were combined with the FISH data to model the annual dynamics of SAR11 subclade populations. 16S rRNA gene clone libraries were constructed to verify the correlation of the T-RFLP data with SAR11 clade structure. Clear vertical and temporal transitions were observed in the dominance of three SAR11 ecotypes. The mechanisms that lead to shifts between the different SAR11 populations are not well understood, but are probably a consequence of finely tuned physiological adaptations that partition the populations along physical and chemical gradients in the ecosystem. The correlation between evolutionary descent and temporal/spatial patterns we describe, confirmed that a minimum of three SAR11 ecotypes occupy the Sargasso Sea surface layer, and revealed new details of their population dynamics. 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At the fine-scale resolution (24 h time series), combining digital PCR and metagenomics, we identified two peaks of cellular infection for the targeted vSAG 37-F6 viral strain (up to approximately 103 copies/ng of prokaryotic DNA), one before sunrise and the second shortly after midday. Considering the high number of co-occurring strains of this microdiverse virus, the abundance values at the species or genus level could be orders of magnitudes higher. These findings represent a significant advancement in understanding the dynamics of the potentially most abundant oceanic virus, providing valuable insights into ecologically relevant marine viruses. C1 [Vila-Nistal, Marina; Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, Carretera San Vicente Raspeig, Alicante 03690, Spain. [Vila-Nistal, Marina; Martinez-Garcia, Manuel] Univ Alicante, Multidisciplinary Inst Environm Studies IMEM, Carretera San Vicente Raspeig, Alicante 03690, Spain. 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Shalchian-Tabrizi, Kamran Bertilsson, Stefan TI Infrequent Transitions between Saline and Fresh Waters in One of the Most Abundant Microbial Lineages (SAR11) SO MOLECULAR BIOLOGY AND EVOLUTION DT Article AB The aquatic bacterial group SAR11 is one of the most abundant organisms on Earth, with an estimated global population size of 2.4 x 10(28) cells in the oceans. Members of SAR11 have also been detected in brackish and fresh waters, but the evolutionary relationships between the species present in the different environments have been ambiguous. In particular, it was not clear how frequently this lineage has crossed the saline-freshwater boundary during its evolutionary diversification. Due to the huge population size of SAR11 and the potential of microbes for long-distance dispersal, we hypothesized that environmental transitions could have occurred repeatedly during the evolutionary diversification of this group. Here, we have constructed extensive 16S rDNA-based molecular phylogenies and undertaken metagenomic data analyses to assess the frequency of saline-freshwater transitions in SAR11 and to investigate the evolutionary implications of this process. Our analyses indicated that very few saline-freshwater transitions occurred during the evolutionary diversification of SAR11, generating genetically distinct saline and freshwater lineages that do not appear to exchange genes extensively via horizontal gene transfer. In contrast to lineages from saline environments, extant freshwater taxa from diverse, and sometimes distant, geographic locations were very closely related. This points to a rapid diversification and dispersal in fresh waters or to slower evolutionary rates in fresh water SAR11 when compared with marine counterparts. In addition, the colonization of both saline and fresh waters appears to have occurred early in the evolution of SAR11. We conclude that the different biogeochemical conditions that prevail in saline and fresh waters have likely prevented the environmental transitions in SAR11, promoting the evolution of clearly distinct lineages in each environment. 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Yau, Stephen S. -T. TI A new method for studying the evolutionary origin of the SAR11 Glade marine bacteria SO MOLECULAR PHYLOGENETICS AND EVOLUTION DT Article AB The free-living SARI I Glade is a globally abundant group of oceanic Alphaproteobacteria, with small genome sizes and rich genomic A+T content. However, the taxonomy of SAR11 has become controversial recently. Some researchers argue that the position of SAR11 is a sister group to Rickettsiales. Other researchers advocate that SAR11 is located within free-living lineages of Alphaproteobacteria. Here, we use the natural vector representation method to identify the evolutionary origin of the SAR11 Glade. This alignment-free method does not depend on any model assumptions. With this approach, the correspondence between proteome sequences and their natural vectors is one-to-one. After fixing a set of proteins, each bacterium is represented by a set of vectors. The Hausdorff distance is then used to compute the dissimilarity distance between two bacteria. The phylogenetic tree can be reconstructed based on these distances. Using our method, we systematically analyze four data sets of alphaproteobacterial proteomes in order to reconstruct the phylogeny of Alphaproteobacteria. From this we can see that the phylogenetic position of the SAR11 group is within a group of other free-living lineages of Alphaproteobacteria. (C) 2016 Elsevier Inc. All rights reserved. C1 [Zhao, Xin; Wan, Xiaogeng; Yau, Stephen S. -T.] Tsinghua Univ, Dept Math Sci, Beijing 100084, Peoples R China. [He, Rong L.] Chicago State Univ, Dept Biol Sci, Chicago, IL 60628 USA. RP Yau, SST (corresponding author), Tsinghua Univ, Dept Math Sci, Beijing 100084, Peoples R China. 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Jones, Leandro R. TI Are ocean currents to slow to counteract SAR11 evolution? A next-generation sequencing, phylogeographic analysis SO MOLECULAR PHYLOGENETICS AND EVOLUTION DT Article AB This work set out to shed light on the phylogeography of the SAR11 Glade of Alphaproteobacteria, which is probably the most abundant group of heterotrophic bacteria on Earth. In particular, we assessed the degree to which empirical evidence (environmental DNA sequences) supports the concept that SAR11 lineages evolve faster than they are dispersed thus generating vicariant distributions, as predicted by recent simulation efforts. We generated 16S rRNA gene sequences from surface seawater collected at the South West Atlantic Ocean and combined these data with previously published sequences from similar environments from elsewhere. Altogether, these data consisted in about 1e6 reads, from which we generated 355,306 high quality sequences of which 95,318 corresponded to SAR11. Quantitative phylogeographic analyses supported the existence of a spatially explicit distribution of SAR11 species and provided evidence in favor of the idea that dispersal limitations significantly contribute to SAR11 radiation throughout the world's oceans. Likewise, pairwise phylogenetic distances between the communities studied here were significantly correlated with the genetic divergences predicted by a previously proposed neutral model. As discussed in the paper, these findings are compatible with the concept that the ocean surface constitutes a homogeneous environment for SAR11, in agreement with previous experimental data. We discuss the implications of this hypothesis in a global change scenario. This is the first study combining high throughput sequencing and phylogenic analysis to study bacterial phylogeography and reporting a distance decay pattern of phylogenetic distances for bacteria. (C) 2016 Elsevier Inc. All rights reserved. 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Lanclos, V. Celeste Faircloth, Brant C. Thrash, J. Cameron TI Cultivation and genomics of the first freshwater SAR11 (LD12) isolate SO ISME JOURNAL DT Article AB Evolutionary transitions between fresh and salt water happen infrequently among bacterioplankton. Within the ubiquitous and highly abundant heterotrophic Alphaproteobacteria order Pelagibacterales (SAR11), most members live in marine habitats, but the LD12 subclade has evolved as a unique freshwater lineage. LD12 cells occur as some of the most dominant freshwater bacterioplankton, yet this group has remained elusive to cultivation, hampering a more thorough understanding of its biology. Here, we report the first successful isolation of an LD12 representative, strain LSUCC0530, using highthroughput dilution-to-extinction cultivation methods, and its complete genome sequence. Growth experiments corroborate ecological data suggesting active populations of LD12 in brackish water up to salinities of similar to 5. 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C1 [Zhao, Yanlin; Qin, Fang; Zhang, Zefeng; Du, Sen] Fujian Agr & Forestry Univ, Fujian Prov Key Lab Agroecol Proc & Safety Monito, Coll Life Sci, Fuzhou, Fujian, Peoples R China. [Zhang, Rui] Xiamen Univ, State Key Lab Marine Environm Sci, Coll Ocean & Earth Sci, Inst Marine Microbes & Ecospheres, Xiamen, Fujian, Peoples R China. [Giovannoni, Stephen J.; Sun, Jing] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Rensing, Christopher] Fujian Agr & Forestry Univ, Fujian Prov Key Lab Soil Environm Hlth & Regulat, Coll Resources & Environm, Fuzhou, Fujian, Peoples R China. [Zhao, Yanlin] Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao, Shandong, Peoples R China. 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Giovannoni, Stephen J. TI SAR11 bacteria have a high affinity and multifunctional glycine betaine transporter SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Marine bacterioplankton face stiff competition for limited nutrient resources. SAR11, a ubiquitous clade of very small and highly abundant Alphaproteobacteria, are known to devote much of their energy to synthesizing ATP-binding cassette periplasmic proteins that bind substrates. We hypothesized that their small size and relatively large periplasmic space might enable them to outcompete other bacterioplankton for nutrients. Using uptake experiments with C-14-glycine betaine, we discovered that two strains of SAR11, Candidatus Pelagibacter sp. HTCC7211 and Cand. P. ubique HTCC1062, have extraordinarily high affinity for glycine betaine (GBT), with half-saturation (K-s) values around 1 nM and specific affinity values between 8 and 14 L mg cell(-1) h(-1). 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Titrations of the algal osmolyte DMSP in seawater medium containing all other macronutrients in excess showed that 1.5 x 10(8) SAR11 cells are produced per nanomole of DMSP. Although it has been shown that other marine alphaproteobacteria use sulphur from DMSP in preference to sulphate(1,2), our results indicate that 'Cand. P. ubique' relies exclusively on reduced sulphur compounds that originate from other plankton. C1 [Tripp, H. James; Kitner, Joshua B.; Schwalbach, Michael S.; Wilhelm, Larry J.; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Dacey, John W. H.] Woods Hole Oceanog Inst, Woods Hole, MA 02543 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, 220 Nash Hall, Corvallis, OR 97331 USA. 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We conducted an analysis of surface ocean waters with the commonly employed hypervariable 4 region SSU rRNA gene primers 515F and 806R, and found that bacteria belonging to the SAR11 clade of Alphaproteobacteria, a group typically making up 20 to 40% of the bacterioplankton in this environment, were greatly underrepresented and comprised <4% of the total community. Using the SILVA reference database, we found a single nucleotide mismatch to nearly all SAR11 subclades, and revised the 806R primer so that it increased the detection of SAR11 clade sequences in the database from 2.6 to 96.7%. We then compared the performance of the original and revised 806R primers in surface seawater samples, and found that SAR11 comprised 0.3 to 3.9% of sequences with the original primers and 17.5 to 30.5% of the sequences with the revised 806R primer. 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Ruiz-Perez, Carlos A. Meziti, Alexandra Hatt, Janet K. Konstantinidis, Konstantinos T. TI Ecogenomic characterization of widespread, closely-related SAR11 clades of the freshwater genus "Candidatus Fonsibacter" and proposal of Ca. Fonsibacter lacus sp. nov SO SYSTEMATIC AND APPLIED MICROBIOLOGY DT Article AB The ubiquitous alpha-proteobacteria of the order "Candidatus Pelagibacterales" (SAR11) are highly abundant in aquatic environments, and among them, members of the monophyletic lineage LD12 (also known as SAR11 Glade IIIb) are specifically found in lacustrine ecosystems. Clade IIIb bacteria are some of the most prominent members of freshwater environments, but little is known about their biology due to the lack of genome representatives. Only recently, the first non-marine isolate was cultured and described as "Candidatus Fonsibacter ubiquis". Here, we expand the collection of freshwater IIIb representatives and describe a new IIIb species of the genus "Ca. Fonsibacter". Specifically, we assembled a collection of 67 freshwater metagenomic datasets from the interconnected lakes of the Chattahoochee River basin (GA, USA) and obtained nearly complete metagenome-assembled genomes (MAGs) representing 5 distinct IIIb subclades, roughly equivalent to species based on genomic standards, including the previously described "Ca. F. ubiquis". Genomic comparisons between members of the IIIb species revealed high similarity in gene content. However, when comparing their abundance profiles in the Chattahoochee basin and various aquatic environments, differences in temporal and spatial distributions among the distinct species were observed implying niche differentiation might be underlying the coexistence of the highly functionally similar representatives. The name Ca. Fonsibacter lacus sp. nov. is proposed for the most abundant and widespread species in the Chattahoochee River basin and various freshwater ecosystems. (C) 2019 Elsevier GmbH. All rights reserved. C1 [Tsementzi, Despina; Rodriguez-R, Luis M.; Meziti, Alexandra; Hatt, Janet K.; Konstantinidis, Konstantinos T.] Georgia Inst Technol, Sch Civil & Environm Engn, Ford Environm Sci & Technol Bldg,311 Ferst Dr, Atlanta, GA 30332 USA. [Ruiz-Perez, Carlos A.; Konstantinidis, Konstantinos T.] Georgia Inst Technol, Sch Biol Sci, Ford Environm Sci & Technol Bldg,311 Ferst Dr, Atlanta, GA 30332 USA. RP Konstantinidis, KT (corresponding author), Georgia Inst Technol, Sch Civil & Environm Engn, 311 Ferst Dr, Atlanta, GA 30332 USA.; Konstantinidis, KT (corresponding author), Georgia Inst Technol, Sch Biol, 311 Ferst Dr, Atlanta, GA 30332 USA. 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Celeste Rasmussen, Anna N. N. Kojima, Conner Y. Y. Cheng, Chuankai Henson, Michael W. W. Faircloth, Brant C. C. Francis, Christopher A. A. Thrash, J. Cameron TI Ecophysiology and genomics of the brackish water adapted SAR11 subclade IIIa SO ISME JOURNAL DT Article AB The Order Pelagibacterales (SAR11) is the most abundant group of heterotrophic bacterioplankton in global oceans and comprises multiple subclades with unique spatiotemporal distributions. Subclade IIIa is the primary SAR11 group in brackish waters and shares a common ancestor with the dominant freshwater IIIb (LD12) subclade. Despite its dominance in brackish environments, subclade IIIa lacks systematic genomic or ecological studies. Here, we combine closed genomes from new IIIa isolates, new IIIa MAGS from San Francisco Bay (SFB), and 460 highly complete publicly available SAR11 genomes for the most comprehensive pangenomic study of subclade IIIa to date. Subclade IIIa represents a taxonomic family containing three genera (denoted as subgroups IIIa.1, IIIa.2, and IIIa.3) that had distinct ecological distributions related to salinity. The expansion of taxon selection within subclade IIIa also established previously noted metabolic differentiation in subclade IIIa compared to other SAR11 subclades such as glycine/serine prototrophy, mosaic glyoxylate shunt presence, and polyhydroxyalkanoate synthesis potential. Our analysis further shows metabolic flexibility among subgroups within IIIa. Additionally, we find that subclade IIIa.3 bridges the marine and freshwater clades based on its potential for compatible solute transport, iron utilization, and bicarbonate management potential. Pure culture experimentation validated differential salinity ranges in IIIa.1 and IIIa.3 and provided detailed IIIa cell size and volume data. 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in the North Sea and characteristics of an abundant RCA isolate SO ISME JOURNAL DT Article AB The Roseobacter group and SAR11 clade constitute high proportions of the marine bacterioplankton, but only scarce information exists on the abundance of distinct populations of either lineage. Therefore, we quantified the abundance of the largest cluster of the Roseobacter group, the RCA (Roseobacter clade affiliated) cluster together with the SAR11 clade by quantitative PCR in the southern and eastern North Sea. The RCA cluster constituted up to 15 and 21% of total bacterial 16S ribosomal RNA (rRNA) genes in September 2005 and May 2006, respectively. At a few stations, the RCA cluster exceeded the SAR11 clade, whereas at most stations, SAR11 constituted higher fractions with maxima of 37%. In most samples, only one RCA ribotype was detected. RCA abundance was positively correlated with phaeopigments, chlorophyll, dissolved and particulate organic carbon (POC), turnover rates of dissolved free amino acids (DFAAs), temperature, and negatively correlated with salinity. The SAR11 clade was only correlated with POC (negatively, May) and with DFAA turnover rates (positively, September). An abundant RCA strain, 'Candidatus Planktomarina temperata', was isolated from the southern North Sea. This strain has an identical 16S rRNA gene sequence to the dominant RCA ribotype. Detection of the pufM gene, coding for a subunit of the reaction center of bacteriochlorophyll a, indicates the potential of the isolate for aerobic anoxygenic photosynthesis. Our study shows that a distinct population of the RCA cluster constitutes an abundant bacterioplankton group in a neritic sea of the temperate zone and indicates that this population has an important role during decaying phytoplankton blooms. The ISME Journal (2011) 5, 8-19; doi:10.1038/ismej.2010.87; published online 1 July 2010 C1 [Giebel, Helge-Ansgar; Kalhoefer, Daniela; Lemke, Andreas; Thole, Sebastian; Gahl-Janssen, Renate; Simon, Meinhard; Brinkhoff, Thorsten] Carl von Ossietzky Univ Oldenburg, Inst Chem & Biol Marine Environm ICBM, D-26111 Oldenburg, Germany. 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A combined set of 2.7 million pyrosequencing reads from the 16S rRNA V1-V2 regions, representing 9 years at the Bermuda Atlantic Time-series Study (BATS) site, was quality checked and parsed into a comprehensive bacterial tree, yielding 929 036 Alphaproteobacteria reads. Phylogenetic structure within the SAR11 clade was linked to seasonally recurring spatiotemporal patterns. This analysis resolved four new SAR11 ecotypes in addition to five others that had been described previously at BATS. The data support a conclusion reached previously that the SAR11 clade diversified by subdivision of niche space in the ocean water column, but the new data reveal a more complex pattern in which deep branches of the clade diversified repeatedly across depth strata and seasonal regimes. The new data also revealed the presence of an unrecognized clade of Alphaproteobacteria, here named SMA-1 (Sargasso Mesopelagic Alphaproteobacteria, group 1), in the upper mesopelagic zone. 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Martin TI The SAR11 Group of Alpha-Proteobacteria Is Not Related to the Origin of Mitochondria SO PLOS ONE DT Article AB Although free living, members of the successful SAR11 group of marine alpha-proteobacteria contain a very small and A+T rich genome, two features that are typical of mitochondria and related obligate intracellular parasites such as the Rickettsiales. Previous phylogenetic analyses have suggested that Candidatus Pelagibacter ubique, the first cultured member of this group, is related to the Rickettsiales+mitochondria clade whereas others disagree with this conclusion. In order to determine the evolutionary position of the SAR11 group and its relationship to the origin of mitochondria, we have performed phylogenetic analyses on the concatenation of 24 proteins from 5 mitochondria and 71 proteobacteria. 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TI Elemental Composition, Phosphorous Uptake, and Characteristics of Growth of a SAR11 Strain in Batch and Continuous Culture SO MSYSTEMS DT Article AB In this study, a strain of SAR11 subgroup Illa (termed HIMB114) was grown in seawater-based batch and continuous culture in order to quantify cellular features and metabolism relevant to SAR11 ecology. We report some of the first direct measurements of cellular elemental quotas for nitrogen (N) and phosphorus (P) for SAR11, grown in batch culture: 1.4 +/- 0.9 fg N and 0.44 +/- 0.01 fg P, respectively, that were consistent with the small size of HIMB114 cells (average volume of 0.09 mu m(3)). However, the mean carbon (C) cellular quota of 50 +/- 47 fg C was anomalously high, but variable. The rates of phosphate (PO43-) uptake measured from both batch and continuous cultures were exceptionally slow: in chemostats growing at 0.3 day(-1), HIMB114 took up 1.1 +/- 0.3 amol P cell day(-1), suggesting that <30% of the cellular P requirement of HIMB114 was met by PO43- assimilation. The mean rate of leucine incorporation, a measure of bacterial production, during late-log-phase growth of batch HIMB114 cultures was 0.042 +/- 0.02 amol Leu cell(-1) h(-1). While only weakly correlated with changes in specific growth rates, the onset of stationary phase resulted in decreases in cell-specific leucine incorporation that were proportional to changes in growth rate. The rates of cellular production, respiratory oxygen consumption, and changes in total organic C concentrations constrained cellular growth efficiencies to 13% +/- 4%. Hence, despite a small genome and diminutively sized cells, SAR11 strain HIMB114 appears to grow at efficiencies similar to those of naturally occurring bacterioplankton communities. IMPORTANCE While SAR11 bacteria contribute a significant fraction to the total picoplankton biomass in the ocean and likely are major players in organic C and nutrient cycling, the cellular characteristics and metabolic features of most lineages have either only been hypothesized from genomes or otherwise not measured in controlled laboratory experimentation. The dearth of data on even the most basic characteristics for what is arguably the most abundant heterotroph in seawater has limited the specific consideration of SAR11 in ocean ecosystem modeling efforts. In this study, we provide measures of cellular P, N, and C, aerobic respiration, and bacterial production for a SAR11 strain growing in natural seawater medium that can be used to directly relate these features of SAR11 to biogeochemical cycling in the oceans. Through the development of a chemostat system to measure nutrient uptake during steady-state growth, we have also documented inorganic P uptake rates that allude to the importance of organic phosphorous to meet cellular P demands, even in the presence of nonlimiting PO43- concentrations. C1 [Grant, Scott R.; Ferron, Sara] Univ Hawaii Manoa, Sch Ocean & Earth Sci & Technol, Dept Oceanog, Honolulu, HI 96822 USA. [Church, Matthew J.] Univ Montana, Flathead Lake Biol Stn, Polson, MT 59860 USA. [Ferron, Sara] Univ Hawaii Manoa, Daniel K Inouye Ctr Microbial Oceanog Res & Educ, Sch Ocean & Earth Sci & Technol, Honolulu, HI 96822 USA. [Laws, Edward A.] Louisiana State Univ, Coll Coast & Environm, Dept Environm Sci, Baton Rouge, LA 70803 USA. [Rappe, Michael S.] Univ Hawaii Manoa, Sch Ocean & Earth Sci & Technol, Hawaii Inst Marine Biol, Kaneohe, HI 96822 USA. RP Rappé, MS (corresponding author), Univ Hawaii Manoa, Sch Ocean & Earth Sci & Technol, Hawaii Inst Marine Biol, Kaneohe, HI 96822 USA. 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Cain, Kelsy R. Hvorecny, Kelli L. Kollman, Justin M. TI Lysogenic host-virus interactions in SAR11 marine bacteria SO NATURE MICROBIOLOGY DT Article AB Here, the authors report the discovery of prophages in cultured SAR11 and show that lysogenic SAR11 produce virions by prophage induction of up to 2.3% of infected cells under carbon-replete growth conditions and up to 30.6% of infected cells under carbon-deplete conditions. Host-virus interactions structure microbial communities, drive biogeochemical cycles and enhance genetic diversity in nature(1,2). Hypotheses proposed to explain the range of interactions that mediate these processes often invoke lysogeny(3-6), a latent infection strategy used by temperate bacterial viruses to replicate in host cells until an induction event triggers the production and lytic release of free viruses. Most cultured bacteria harbour temperate viruses in their genomes (prophage)(7). The absence of prophages in cultures of the dominant lineages of marine bacteria has contributed to an ongoing debate over the ecological significance of lysogeny and other viral life strategies in nature(6,8-15). Here, we report the discovery of prophages in cultured SAR11, the ocean's most abundant clade of heterotrophic bacteria(16,17). We show the concurrent production of cells and viruses, with enhanced virus production under carbon-limiting growth conditions. Evidence that related prophages are broadly distributed in the oceans suggests that similar interactions have contributed to the evolutionary success of SAR11 in nutrient-limited systems. C1 [Morris, Robert M.; Cain, Kelsy R.] Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. [Hvorecny, Kelli L.; Kollman, Justin M.] Univ Washington, Dept Biochem, Seattle, WA 98195 USA. RP Morris, RM (corresponding author), Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. 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In freshwater lakes, the clade is represented by tribe LD12 which is phylogenetically distinct from the marine SAR11. We studied the ecology of LD12 in a temperate dimictic lake (Lake Erken, Sweden), by analyzing its seasonal dynamics with quantitative PCR, CARD-FISH and 454 pyrosequencing of the 16S rRNA gene. Results showed that LD12 can be as numerous in freshwater bacterioplankton as their marine SAR11 siblings. They exhibited strong seasonality and made up from 1.8 to 40% of the total bacterial 16S rRNA pool (mean 14%) with pronounced peaks in summer and late fall. Except in spring, LD12 was the dominant Alphaproteobacteria, contributing on average 72% of the 16S rRNA within this class. The LD12 population was dominated by a single persistent ribotype, suggesting low local divergence, at least at the phylogenetic resolution accessed with rRNA genes. The relative abundance of LD12 was positively correlated to nutrient concentrations (phosphate, ammonia, nitrate, and silica) and water transparency whereas the relative abundance was lower during periods characterized by high phytoplankton biomass. Based on these observations we propose that LD12 are poor competitors during periods of high phytoplankton productivity and associated release of labile organic compounds, but thrive when availability of inorganic nutrients is high. Similar to the marine SAR11 sibling group, local LD12 populations appear to respond in contrasting ways to nutrient availability in different lakes, pointing to either ecological divergence within the tribe or variations in the interplay between environmental driver variables. C1 [Bertilsson, Stefan] Uppsala Univ, Dept Ecol & Genet, S-75236 Uppsala, Sweden. Uppsala Univ, Limnol Lab, S-75236 Uppsala, Sweden. Uppsala Univ, Sci Life Lab, S-75236 Uppsala, Sweden. 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Hogle, Shane L. Rosendo, Kali Chisholm, Sallie W. TI Co-culture and biogeography of Prochlorococcus and SAR11 SO ISME JOURNAL DT Article AB Prochlorococcus and SAR11 are among the smallest and most abundant organisms on Earth. With a combined global population of about 2.7 x 10(28) cells, they numerically dominate bacterioplankton communities in oligotrophic ocean gyres and yet they have never been grown together in vitro. Here we describe co-cultures of Prochlorococcus and SAR11 isolates representing both high- and low-light adapted clades. We examined: (1) the influence of Prochlorococcus on the growth of SAR11 and vice-versa, (2) whether Prochlorococcus can meet specific nutrient requirements of SAR11, and (3) how co-culture dynamics vary when Prochlorococcus is grown with SAR11 compared with sympatric copiotrophic bacteria. SAR11 grew 15-70% faster in co-culture with Prochlorococcus, while the growth of the latter was unaffected. When Prochlorococcus populations entered stationary phase, this commensal relationship rapidly became amensal, as SAR11 abundances decreased dramatically. In parallel experiments with copiotrophic bacteria; however, the heterotrophic partner increased in abundance as Prochlorococcus densities leveled off. The presence of Prochlorococcus was able to meet SAR11's central requirement for organic carbon, but not reduced sulfur. Prochlorococcus strain MIT9313, but not MED4, could meet the unique glycine requirement of SAR11, which could be due to the production and release of glycine betaine by MIT9313, as supported by comparative genomic evidence. Our findings also suggest, but do not confirm, that Prochlorococcus MIT9313 may compete with SAR11 for the uptake of 3-dimethylsulfoniopropionate (DMSP). 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Todd, Jonathan D. Johnston, Andrew W. B. TI Manganese uptake in marine bacteria; the novel MntX transporter is widespread in Roseobacters, Vibrios, Alteromonadales and the SAR11 and SAR116 clades SO ISME JOURNAL DT Article AB We showed that two very different manganese transporters occur in various important genera of marine bacteria. The ABC transporter encoded by sitABCD of the model Roseobacter-clade bacterium Ruegeria pomeroyi DSS-3 is required for Mn2+ import and was repressed by the Mur (Manganese uptake regulator) transcriptional regulator in Mn-replete media. Most genome-sequenced Roseobacter strains contain SitABCD, which are in at least two sub-groups, judged by their amino-acid sequences. However, a few Roseobacters, for example, Roseovarius nubinhibens, lack sitABCD, but these contain another gene, mntX, which encodes a predicted inner membrane polypeptide and is preceded by cis-acting Mur-responsive MRS sequences. It was confirmed directly that mntX of Roseovarius nubinhibens encodes a manganese transporter that was required for growth in Mn-depleted media and that its expression was repressed by Mur in Mn-replete conditions. MntX homologues occur in the deduced proteomes of several bacterial species. Strikingly, all of these live in marine habitats, but are in distantly related taxonomic groups, in the gamma-and alpha-proteobacteria. Notably, MntX was prevalent in nearly all strains of Vibrionales, including the important pathogen, Vibrio cholerae. It also occurs in a strain of the hugely abundant Candidatus Pelagibacter (SAR11), and in another populous marine bacterium, Candidatus Puniceispirillum marinum (SAR116). Consistent with this, MntX was abundant in marine bacterial metagenomes, with one sub-type occurring in an as-yet unknown bacterial clade. 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Haroon, Mohamed F. Shibl, Ahmed A. Cahill, Matt J. Ngugi, David K. Williams, Gareth J. Morton, James T. Knight, Rob Goodwin, Kelly D. Stingl, Ulrich TI Red Sea SAR11 and Prochlorococcus Single-Cell Genomes Reflect Globally Distributed Pangenomes SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Evidence suggests many marine bacteria are cosmopolitan, with widespread but sparse strains poised to seed abundant populations under conducive growth conditions. However, studies supporting this "microbial seed bank" hypothesis have analyzed taxonomic marker genes rather than whole genomes/metagenomes, leaving open the possibility that disparate ocean regions harbor endemic gene content. The Red Sea is isolated geographically from the rest of the ocean and has a combination of high irradiance, high temperature, and high salinity that is unique among the oceans; we therefore asked whether it harbors endemic gene content. We sequenced and assembled single-cell genomes of 21 SAR11 (subclades Ia, Ib, Id, and II) and 5 Prochlorococcus (ecotype HLII) samples from the Red Sea and combined them with globally sourced reference genomes to cluster genes into ortholog groups (OGs). Ordination of OG composition could distinguish clades, including phylogenetically cryptic Prochlorococcus ecotypes LLII and LLIII. Compared with reference genomes, 1% of Prochlorococcus and 17% of SAR11 OGs were unique to the Red Sea genomes (RS-OGs). Most (83%) RS-OGs had no annotated function, but 65% of RS-OGs were expressed in diel Red Sea metatranscriptomes, suggesting they are functional. Searching Tara Oceans metagenomes, RS-OGs were as likely to be found as non-RS-OGs; nevertheless, Red Sea and other warm samples could be distinguished from cooler samples using the relative abundances of OGs. The results suggest that the prevalence of OGs in these surface ocean bacteria is largely cosmopolitan, with differences in population metagenomes manifested by differences in relative abundance rather than complete presence/absence of OGs. IMPORTANCE Studies have shown that as we sequence seawater from a selected environment deeper and deeper, we approach finding every bacterial taxon known for the ocean as a whole. However, such studies have focused on taxonomic marker genes rather than on whole genomes, raising the possibility that the lack of endemism results from the method of investigation. We took a geographically isolated water body, the Red Sea, and sequenced single cells from it. We compared those single-cell genomes to available genomes from around the ocean and to ocean-spanning metagenomes. We showed that gene ortholog groups found in Red Sea genomes but not in other genomes are nevertheless common across global ocean metagenomes. 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[Morton, James T.; Knight, Rob] Univ Calif San Diego, Dept Comp Sci & Engn, La Jolla, CA 92093 USA. [Knight, Rob] Univ Calif San Diego, Ctr Microbiome Innovat, La Jolla, CA 92093 USA. [Stingl, Ulrich] Univ Florida, Dept Microbiol & Cell Sci, Ft Lauderdale Res & Educ Ctr, UF Inst Food & Agr Sci, Davie, FL USA. [Shibl, Ahmed A.] NYU, Marine Microbial Ecol Lab, Biol Program, Abu Dhabi, U Arab Emirates. RP Thompson, LR; Stingl, U (corresponding author), King Abdullah Univ Sci & Technol, Red Sea Res Ctr, Thuwal, Saudi Arabia.; Thompson, LR (corresponding author), Univ Southern Mississippi, Dept Biol Sci, Hattiesburg, MS 39406 USA.; Thompson, LR (corresponding author), Univ Southern Mississippi, Northern Gulf Inst, Hattiesburg, MS 39406 USA. 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Rodriguez-Valera, Francisco Martinez-Garcia, Manuel TI Single-cell genomics uncover Pelagibacter as the putative host of the extremely abundant uncultured 37-F6 viral population in the ocean SO ISME JOURNAL DT Article AB The identification of relevant virus host pairs that globally account for a large pool of carbon and nutrients in the ocean is paramount to build accurate ecological models. A previous work using single-virus genomics led to the discovery of the uncultured single-virus vSAG 37-F6, originally sorted from the Mediterranean Sea (Blanes Bay Microbial Observatory), that represents one of the most abundant dsDNA viral population in the marine surface virosphere. Here, from same sampling site, we report that a Pelagibacter single-cell contained a viral member of vSAG 37-F6 population, by means of PCR screening of sorted, genome-amplified single cells with vSAG 37-F6-specific primers and whole-genome sequencing. Furthermore, viruses from this population were also found in three other Pelagibacter single cells from the South Pacific and Atlantic oceans. These new uncultured pelagiphages were genetically different from the previously characterized pelagiphage isolates. Data showed that the uncultured vSAG 37-F6 population represents the Pelagibacter phages that inhabit the sunlit ocean better, and contains a vast unrecognized microdiversity. C1 [Martinez-Hernandez, Francisco; Lluesma Gomez, Monica; Garcia-Heredia, Inmaculada; Maestre-Carballa, Lucia; Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. [Fornas, Oscar] Pompeu Fabra Univ, Flow Cytometry Unit, Barcelona, Spain. [Fornas, Oscar] Barcelona Inst Sci & Technol, Ctr Genom Regulat, Barcelona, Spain. [Lopez-Perez, Mario; Haro-Moreno, Jose M.; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Evolutionary Genom Grp, Div Microbiol, Apartado 18, Alicante 03550, Spain. RP Martinez-Garcia, M (corresponding author), Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. 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Carbon Pump Ocean, P54, DOI DOI 10.1126/SCIENCE.OPMS.SB0001 Zhao YL, 2013, NATURE, V494, P357, DOI 10.1038/nature11921 NR 26 TC 31 Z9 36 PD JAN PY 2019 VL 13 IS 1 BP 232 EP 236 DI 10.1038/s41396-018-0278-7 UT WOS:000453576600019 DA 2025-07-30 ER PT J AU Viklund, J Martijn, J Ettema, TJG Andersson, SGE AF Viklund, Johan Martijn, Joran Ettema, Thijs J. G. Andersson, Siv G. E. TI Comparative and Phylogenomic Evidence That the Alphaproteobacterium HIMB59 Is Not a Member of the Oceanic SAR11 Clade SO PLOS ONE DT Article AB SAR11 is a globally abundant group of Alphaproteobacteria in the oceans that is taxonomically not well defined. It has been suggested SAR11 should be classified into the novel order Pelagibacterales. Features such as conservation of gene content and synteny have been taken as evidence that also the divergent member HIMB59 should be included in the order. However, this proposition is controversial since phylogenetic analyses have questioned the monophyly of this grouping. Here, we performed phylogenetic analyses and reinvestigated the genomic similarity of SAR11 and HIMB59. Our phylogenetic analysis confirmed that HIMB59 is not a sister group to the other SAR11 strains. By placing the comparison in the context of the evolution of the Alphaproteobacteria, we found that none of the measures of genomic similarity supports a clustering of HIMB59 and SAR11 to the exclusion of other Alphaproteobacteria. First, pairwise sequence similarity measures for the SAR11 and HIMB59 genomes were within the range observed for unrelated pairs of Alphaproteobacteria. Second, pairwise comparisons of gene contents revealed a higher similarity of SAR11 to several other alphaproteobacterial genomes than to HIMB59. Third, the SAR11 genomes are not more similar in gene order to the HIMB59 genome than what they are to several other alphaproteobacterial genomes. Finally, in contrast to earlier reports, we observed no sequence similarity between the hypervariable region HVR2 in the SAR11 genomes and the region located at the corresponding position in the HIMB59 genome. Based on these observations, we conclude that the alphaproteobacterium HIMB59 is not monophyletic with the SAR11 strains and that genome streamlining has evolved multiple times independently in Alphaproteobacteria adapted to the upper surface waters of the oceans. C1 [Viklund, Johan; Martijn, Joran; Ettema, Thijs J. G.; Andersson, Siv G. E.] Biomed Ctr, Dept Mol Evolut, Sci Life Lab, Uppsala, Sweden. RP Andersson, SGE (corresponding author), Biomed Ctr, Dept Mol Evolut, Sci Life Lab, Uppsala, Sweden. 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TI Improvements of high-throughput culturing yielded novel SAR11 strains and other abundant marine bacteria from the Oregon coast and the Bermuda Atlantic Time Series study site SO ISME JOURNAL DT Article AB The introduction of high-throughput dilution-to-extinction culturing (HTC) of marine bacterioplankton using sterilized natural sea water as media yielded isolates of many abundant but previously uncultured marine bacterial clades. In early experiments, bacteria from the SAR11 cluster (class Alphaproteobacteria), which are presumed to be the most abundant prokaryotes on earth, were cultured. Although many additional attempts were made, no further strains of the SAR11 clade were obtained. Here, we describe improvements to the HTC technique, which led to the isolation of 17 new SAR11 strains from the Oregon coast and the Sargasso Sea, accounting for 28% and 31% of all isolates in these experiments. Phylogenetic analysis of the internal transcribed spacer (ITS) region showed that the isolates from the Oregon coast represent three different subclusters of SAR11, while isolates from the Sargasso Sea were more uniform and represented a single ITS cluster. A PCR assay proved the presence of proteorhodopsin (PR) in nearly all SAR11 isolates. Analysis of PR amino-acid sequences indicated that isolates from the Oregon coast were tuned to either green or blue light, while PRs from strains obtained from the Sargasso Sea were exclusively tuned to maximum absorbance in the blue. Interestingly, phylogenies based on PR and ITS did not correlate, suggesting lateral gene transfer. In addition to the new SAR11 strains, many novel strains belonging to clusters of previously uncultured or undescribed species of different bacterial phyla, including the first strain of the highly abundant alphaproteobacterial SAR116 clade, were isolated using the modified methods. C1 Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. RP Stingl, U (corresponding author), Oregon State Univ, Dept Microbiol, 220 Nash Hall, Corvallis, OR 97331 USA. 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Bertagnolli, Anthony D. Tsementzi, Despina Woyke, Tanja Stewart, Frank J. Konstantinidis, Konstantinos T. TI Description of Candidatus Mesopelagibacter carboxydoxydans and Candidatus Anoxipelagibacter denitrificans: Nitrate-reducing SAR11 genera that dominate mesopelagic and anoxic marine zones SO SYSTEMATIC AND APPLIED MICROBIOLOGY DT Article AB The diverse and ubiquitous members of the SAR11 lineage (Alphaproteobacteria) represent up to 30-40% of the surface and mesopelagic oceanic microbial communities. However, the molecular and ecological mechanisms that differentiate closely related, yet distinct, SAR11 members that often co-occur under similar environmental conditions remain speculative. Recently, two mesopelagic and oxygen minimum zone (OMZ)-associated subclades of SAR11 (Ic and IIa.A) were described using single-cell amplified genomes (SAGs) linked to nitrate reduction in OMZs. In this current study, the collection of genomes belonging to these two subclades was expanded with thirteen new metagenome-assembled genomes (MAGs), thus providing a more detailed phylogenetic and functional characterization of these subclades. Gene content based predictions of metabolic functions revealed similarities in central carbon metabolism between subclades Ic and IIa.A and surface SAR11 clades, with small variations in central pathways. These variations included more versatile sulfur assimilation pathways, as well as a previously predicted capacity for nitrate reduction that conferred unique versatility on mesopelagic-adapted clades compared to their surface counterparts. Finally, consistent with previously reported abundances of carbon monoxide (CO) in surface and mesopelagic waters, subclades Ia (surface) and Ic (mesopelagic) have the genetic potential to oxidize carbon monoxide (CO), presumably taking advantage of this abundant compound as an electron donor. Based on genomic analyses, environmental distribution and metabolic reconstruction, we propose two new SAR11 genera, Ca. Mesopelagibacter carboxydoxydans (subclade Ic) and Ca. Anoxipelagibacter denitrificans (subclade IIa.A), which represent members of the mesopelagic and OMZ-adapted SAR11 clades. (c) 2021 Elsevier GmbH. All rights reserved. C1 [Ruiz-Perez, Carlos A.; Bertagnolli, Anthony D.; Stewart, Frank J.; Konstantinidis, Konstantinos T.] Georgia Inst Technol, Sch Biol Sci, Atlanta, GA 30332 USA. [Tsementzi, Despina; Konstantinidis, Konstantinos T.] Georgia Inst Technol, Sch Civil & Environm Engn, Atlanta, GA 30332 USA. [Woyke, Tanja] DOE Joint Genome Inst, One Cyclotron Rd,Mail Stop 91R0183, Berkeley, CA 94720 USA. [Stewart, Frank J.] Georgia Inst Technol, Ctr Microbial Dynam & Infect, Atlanta, GA 30332 USA. [Stewart, Frank J.] Montana State Univ, Dept Microbiol & Immunol, Bozeman, MT 59717 USA. [Konstantinidis, Konstantinos T.] Georgia Inst Technol, Ctr Bioinformat & Computat Genom, Atlanta, GA 30332 USA. RP Konstantinidis, KT (corresponding author), Georgia Inst Technol, Sch Civil & Environm Engn, Atlanta, GA 30332 USA. 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Halsey, Kimberly H. Saw, Jimmy Muslin, Omran Suffridge, Christopher P. Sun, Jing Lee, Chih-Ping Moore, Eric R. Temperton, Ben Noell, Stephen E. TI A Parasitic Arsenic Cycle That Shuttles Energy from Phytoplankton to Heterotrophic Bacterioplankton SO MBIO DT Article AB In many regions of the world oceans, phytoplankton face the problem of discriminating between phosphate, an essential nutrient, and arsenate, a toxic analogue. Many phytoplankton, including the most abundant phytoplankton group known, Prochlorococcus, detoxify arsenate (AsV) by reduction to arsenite (AsIII), followed by methylation and excretion of the methylated arsenic products. We synthesized [C-14]dimethyl arsenate (DMA) and used it to show that cultured Pelagibacter strain HTCC7211 (SAR11) cells oxidize the methyl group carbons of DMA, producing (CO2)-C-14 and ATP. We measured [C-14] DMA oxidation rates in the P-depleted surface waters of the Sargasso Sea, a subtropical ocean gyre. [C-14]DMA was oxidized to (CO2)-C-14 by Sargasso Sea plankton communities at a rate that would cause turnover of the estimated DMA standing stock every 8.1 days. SAR11 strain HTCC7211, which was isolated from the Sargasso Sea, has a pair of arsenate resistance genes and was resistant to arsenate, showing no growth inhibition at As/P ratios of >65:1. Across the global oceans, there was a strong inverse relationship between the frequency of the arsenate reductase (LMWPc_ArsC) in Pelagibacter genomes and phosphate concentrations. We propose that the demethylation of methylated arsenic compounds by Pelagibacter and possibly other bacterioplankton, coupled with arsenate resistance, results in the transfer of energy from phytoplankton to bacteria. We dub this a parasitic cycle because the release of arsenate by Pelagibacter in principle creates a positive-feedback loop that forces phytoplankton to continually regenerate arsenate detoxification products, producing a flow of energy to P-limited ocean regions. IMPORTANCE In vast, warm regions of the oceans, phytoplankton face the problem of arsenic poisoning. Arsenate is toxic because it is chemically similar to phosphate, a scarce nutrient that phytoplankton cells need for growth. Many phytoplankton, including the commonest phytoplankton type in warm oceans, Prochlorococcus, detoxify arsenate by adding methyl groups. Here we show that the most abundant non-photosynthetic plankton in the oceans, SAR11 bacteria, remove the methyl groups, releasing poisonous forms of arsenic back into the water. We postulate that the methylation and demethylation of arsenic compounds creates a cycle in which the phytoplankton can never get ahead and must continually transfer energy to the SAR11 bacteria. We dub this a parasitic process and suggest that it might help explain why SAR11 bacteria are so successful, surpassing all other plankton in their numbers. Field experiments were done in the Sargasso Sea, a subtropical ocean gyre that is sometimes called an ocean desert because, throughout much of the year, there is not enough phosphorous in the water to support large blooms of phytoplankton. Ocean deserts are expanding as the oceans absorb heat and grow warmer. C1 [Giovannoni, Stephen J.; Halsey, Kimberly H.; Saw, Jimmy; Muslin, Omran; Suffridge, Christopher P.; Sun, Jing; Lee, Chih-Ping; Moore, Eric R.; Temperton, Ben; Noell, Stephen E.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Saw, Jimmy] George Washington Univ, Washington, DC USA. 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Helge-Ansgar Rink, Beate Grayek, Sebastian Staneva, Joanna Stanev, Emil Simon, Meinhard TI Differential effects of hydrographic and biogeo chemical properties on the SAR11 clade and Roseobacter RCA cluster in the North Sea SO AQUATIC MICROBIAL ECOLOGY DT Article AB The SAR11 clade and the Roseobacter clade affiliated (RCA) cluster belong to the most prominent bacterioplankton groups in temperate to polar seas. Despite some insights into biological controls of both lineages, little is known about environmental, hydrographic and biogeochemical controls. Therefore, we assessed the abundance of both lineages using quantitative PCR in the southern North Sea, subjected them to a multiple linear regression analysis and related their occurrence to current patterns by backtracking the water masses found at individual stations for the preceding 24 to 27 d. SAR11 constituted <1 to 47% of total bacterial 16S rRNA genes. The abundance of this clade was inversely correlated to the salinity change of the water masses at the stations, indicating a preference for stable and presumably nutrient depleted waters. The RCA cluster constituted <1 to 5% of total bacterial 16S rRNA genes but did not exhibit any correlation to hydrographic properties. 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Hehenberger, Elisabeth Giovannoni, Stephen J. Worden, Alexandra Z. TI Genomes from Uncultivated Pelagiphages Reveal Multiple Phylogenetic Clades Exhibiting Extensive Auxiliary Metabolic Genes and Cross-Family Multigene Transfers SO MSYSTEMS DT Article AB For the abundant marine Alphaproteobacterium Pelagibacter (SARI 1), and other bacteria, phages are powerful forces of mortality. However, little is known about the most abundant Pelagiphages in nature, such as the widespread HTVC023P-type, which is currently represented by two cultured phages. Using viral metagenomic data sets and fluorescence-activated cell sorting, we recovered 80 complete, undescribed Podoviridae genomes that form 10 phylogenomically distinct clades (herein, named Clades I to X) related to the HTVC023P-type. These expanded the HTVC023P-type pangenome by 15-fold and revealed 41 previously unknown auxiliary metabolic genes (AMGs) in this viral lineage. Numerous instances of partner-AMGs (colocated and involved in related functions) were observed, including partners in nucleotide metabolism, DNA hypermodification, and Curli biogenesis. The Type VIII secretion system (T8SS) responsible for Curli biogenesis was identified in nine genomes and expanded the repertoire of T8SS proteins reported thus far in viruses. Additionally, the identified T8SS gene cluster contained an iron-dependent regulator (FecR), as well as a histidine kinase and adenylate cyclase that can be implicated in T8SS function but are not within T8SS operons in bacteria. While T8SS are lacking in known Pelagibacter, they contribute to aggregation and biofilm formation in other bacteria. Phylogenetic reconstructions of partner-AMGs indicate derivation from cellular lineages with a more recent transfer between viral families. For example, homologs of all T8SS genes are present in syntenic regions of distant Myoviridae Pelagiphages, and they appear to have alphaproteobacterial origins with a later transfer between viral families. The results point to an unprecedented multipartner-AMG transfer between marine Myoviridae and Podoviridae. Together with the expansion of known metabolic functions, our studies provide new prospects for understanding the ecology and evolution of marine phages and their hosts. IMPORTANCE One of the most abundant and diverse marine bacterial groups is Pelagibacter. Phages have roles in shaping Pelagibacter ecology; however, several Pelagiphage lineages are represented by only a few genomes. This paucity of data from even the most widespread lineages has imposed limits on the understanding of the diversity of Pelagiphages and their impacts on hosts. 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Rodriguez-Valera, Francisco Lopez-Perez, Mario TI Extensive paralogism in the environmental pangenome: a key factor in the ecological success of natural SAR11 populations SO MICROBIOME DT Article AB Background The oceanic microbiome is dominated by members of the SAR11 clade. Despite their abundance, challenges in recovering the full genetic diversity of natural populations have hindered our understanding of the eco-evolutionary mechanisms driving intra-species variation. In this study, we employed a combination of single-amplified genomes and long-read metagenomics to recover the genomic diversity of natural populations within the SAR11 genomospecies Ia.3/VII, the dominant group in the Mediterranean Sea. Results The reconstruction of the first complete genome within this genomospecies revealed that the core genome represents a significant proportion of the genome (similar to 81%), with highly divergent areas that allow for greater strain-dependent metabolic flexibility. The flexible genome was concentrated in small regions, typically containing a single gene, and was located in equivalent regions within the genomospecies. Each variable region was associated with a specific set of genes that, despite exhibiting some divergence, maintained equivalent biological functionality within the population. The environmental pangenome is large and enriched in genes involved in nutrient transport, as well as cell wall synthesis and modification, showing an extremely high degree of functional redundancy in the flexible genome (i.e. paralogisms). Conclusions This genomic architecture promotes polyclonality, preserving genetic variation within the population. This, in turn, mitigates intraspecific competition and enables the population to thrive under variable environmental conditions and selective pressures. 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In this study, despite seasonal fluctuations, several bacterial populations (<2% of the total OTUs) that are highly dominant (25% of relative abundance) and highly frequently occurring (>85% of occurrence) over four seasons were identified. Mathematical models using occurrence frequency and relative abundance data were able to describe community assembly persisting over time. Further, this work uncovered one of the core bacterial populations phylogenetically affiliated to SAR11 subclade Ma. The analysis of the hypervariable region of 16S rRNA gene and single copy housekeeping genes recovered from metagenomic datasets suggested that the population was unexpectedly evolutionarily closely related to those inhabiting a mesosaline lacustrine ecosystem rather than other marine/coastal members. 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James Schwalbach, Michael S. Meyer, Michelle M. Kitner, Joshua B. Breaker, Ronald R. Giovannoni, Stephen J. TI Unique glycine-activated riboswitch linked to glycine-serine auxotrophy in SAR11 SO ENVIRONMENTAL MICROBIOLOGY DT Article AB The genome sequence of the marine bacterium 'Candidatus Pelagibacter ubique' and subsequent analyses have shown that while it has a genome as small as many obligate parasites, it nonetheless possesses a metabolic repertoire that allows it to grow as one of the most successful free-living cells in the ocean. An early report based on metabolic reconstruction indicated that SAR11 cells are prototrophs for all amino acids. However, here we report experimental evidence that 'Cand. P. ubique' is effectively auxotrophic for glycine and serine. With glucose and acetate added to seawater to supply organic carbon, the addition of 125 nM to 1.5 mu M glycine to growth medium containing all other nutrients in excess resulted in a linear increase in maximum cell density from 1.14 x 10(6) cells ml(-1) to 8.16 x 10(6) cells ml(-1) (R-2 = 0.992). Serine was capable of substituting for glycine at 1.5 mu M. 'Cand. P. ubique' contains a glycine-activated riboswitch preceding malate synthase, an unusual genomic context that is conserved in the SAR11 group. Malate synthase plays a critical role in central metabolism by enabling TCA intermediates to be regenerated through the glyoxylate cycle. In vitro analysis of this riboswitch indicated that it responds solely to glycine but not close structural analogues, such as glycine betaine, malate, glyoxylate, glycolate, alanine, serine or threonine. We conclude that 'Cand. 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Here, we have identified and cloned two putative epoxide hydrolase genes (cpeh and sneh) from marine bacteria, Candidatus pelagibacter ubique and terrestrial bacteria, Stackebrandtia nassauensis, respectively and overexpressed them in pET28a vector in Escherichia coli BL21(DE3). The CPEH protein (42 kDa) was found to be overexpressed as inactive inclusion bodies while SNEH protein (40 kDa) was found to form soluble aggregates. In this study, the recombinant CPEH was successfully transformed from insoluble aggregates to the soluble and functionally active form, using pCold TF vector, though with low EH activity. To prevent the soluble aggregate formation of SNEH, it was co-expressed with GroEL/ES chaperone and was also fused with trigger factor (TF) chaperone at its N-terminus. The TF chaperone-assisted correct folding of SNEH led to a purified active EH with a specific activity of 3.85 mu mol/min/mg. 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Hartmann, Manuela Grob, Carolina Tarran, Glen A. Martin, Adrian P. Fuchs, Bernhard M. Scanlan, David J. Zubkov, Mikhail V. TI Comparable light stimulation of organic nutrient uptake by SAR11 and Prochlorococcus in the North Atlantic subtropical gyre SO ISME JOURNAL DT Article AB Subtropical oceanic gyres are the most extensive biomes on Earth where SAR11 and Prochlorococcus bacterioplankton numerically dominate the surface waters depleted in inorganic macronutrients as well as in dissolved organic matter. In such nutrient poor conditions bacterioplankton could become photoheterotrophic, that is, potentially enhance uptake of scarce organic molecules using the available solar radiation to energise appropriate transport systems. Here, we assessed the photoheterotrophy of the key microbial taxa in the North Atlantic oligotrophic gyre and adjacent regions using P-33-ATP, H-3-ATP and S-35-methionine tracers. Light-stimulated uptake of these substrates was assessed in two dominant bacterioplankton groups discriminated by flow cytometric sorting of tracer-labelled cells and identified using catalysed reporter deposition fluorescence in situ hybridisation. One group of cells, encompassing 48% of all bacterioplankton, were identified as members of the SAR11 clade, whereas the other group (24% of all bacterioplankton) was Prochlorococcus. When exposed to light, SAR11 cells took 31% more ATP and 32% more methionine, whereas the Prochlorococcus cells took 33% more ATP and 34% more methionine. Other bacterioplankton did not demonstrate light stimulation. Thus, the SAR11 and Prochlorococcus groups, with distinctly different light-harvesting mechanisms, used light equally to enhance, by approximately one-third, the uptake of different types of organic molecules. 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Methodology/Principal Findings: We present a phylogenetic study of the origin of mitochondria that incorporates metagenome data from the Global Ocean Sampling (GOS) expedition. We identify mitochondrially related sequences in the GOS dataset that represent a rare group of Alphaproteobacteria, designated OMAC (Oceanic Mitochondria Affiliated Clade) as the closest free-living relatives to mitochondria in the oceans. In addition, our analyses reject the hypothesis that the mitochondrial system for aerobic respiration is affiliated with that of the SAR11 clade. Conclusions/Significance: Our results allude to the existence of an alphaproteobacterial clade in the oxygen-rich surface waters of the oceans that represents the closest free-living relative to mitochondria identified thus far. In addition, our findings underscore the importance of expanding the taxonomic diversity in phylogenetic analyses beyond that represented by cultivated bacteria to study the origin of mitochondria. 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Hyde, Emily Leao, Pedro TI Nature should be the model for microbial sciences SO JOURNAL OF BACTERIOLOGY DT Review AB Until recently, microbiologists have relied on cultures to understand the microbial world. As a result, model organisms have been the focus of research into understanding Bacteria and Archaea at a molecular level. Diversity surveys and metagenomic sequencing have revealed that these model species are often present in low abundance in the environment; instead, there are microbial taxa that are cosmopolitan in nature. Due to the numerical dominance of these microorganisms and the size of their habitats, these lineages comprise mind-boggling population sizes upward of 10(28) cells on the planet. Many of these dominant groups have cultured representatives and have been shown to be involved in mediating key processes in nature. Given their importance and the increasing need to understand changes due to climate change, we propose that members of Nitrosophaerota (Nitrosopumilus maritimus), SAR11 (Pelagibacter ubique), Hadesarchaeia, Bathyarchaeia, and others become models in the future. Abundance should not be the only measure of a good model system; there are other organisms that are well suited to advance our understanding of ecology and evolution. For example, the most well-studied symbiotic bacteria, like Buchnera, Aliivibrio, and Rhizobium, should be models for understanding host-associations. Also, there are organisms that hold new insights into major transitions in the evolution of life on the planet like the Asgard Archaea (Heimdallarchaeia). Innovations in a variety of in situ techniques have enabled us to circumvent culturing when studying everything from genetics to physiology. Our deepest understanding of microbiology and its impact on the planet will come from studying these microbes in nature. 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Moran, Mary Ann Whitman, William B. TI Bacterial catabolism of dimethylsulfoniopropionate (DMSP) SO FRONTIERS IN MICROBIOLOGY DT Review AB Dimethylsulfoniopropionate (DMSP) is a metabolite produced primarily by marine phytoplankton and is the main precursor to the climatically important gas dimethylsulfide (DMS). DMS is released upon bacterial catabolism of DMSP but it is not the only possible fate of DMSP sulfur. An alternative demethylation/demethiolation pathway results in the eventual release of methanethiol, a highly reactive volatile sulfur compound that contributes little to the atmospheric sulfur flux. The activity of these pathways control the natural flux of sulfur released to the atmosphere. Although these biochemical pathways and the factors that regulate them are of great interest, they are poorly understood. Only recently have some of the genes and pathways responsible for DMSP catabolism been elucidated. Thus far, six different enzymes have been identified that catalyze the cleavage of DMSP resulting in the release of DMS. In addition, five of these enzymes appear to produce acrylate, while one produces 3-hydroxypropionate. In contrast, only one enzyme, designated DmdA, has been identified that catalyzes the demethylation reaction producing methylmercaptopropionate (MMPA). The metabolism of MMPA is performed by a series of three coenzyme-A mediated reactions catalyzed by DmdB, DmdC, and DmdD. Interestingly, Candidatus Pelagibacter ubique, a member of the SAR11 clade of Alphaproteobacteria that is highly abundant in marine surface waters, possessed functional DmdA, DmdB, and DmdC enzymes. Microbially mediated transformations of both DMS and methanethiol are also possible, although many of the biochemical and molecular genetic details are still unknown. 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Xia, Qian Zhao, Yanlin TI Genomic diversity, life strategies and ecology of marine HTVC010P-type pelagiphages SO MICROBIAL GENOMICS DT Article AB SAR11 bacteria dominate ocean surface bacterioplankton communities, and play an important role in marine carbon and nutrient cycling. The biology and ecology of SAR11 are impacted by SAR11 phages (pelagiphages) that are highly diverse and abundant in the ocean. Among the currently known pelagiphages, HTVC010P represents an extremely abundant but under- studied phage group in the ocean. In this study, we have isolated seven new HTVC010P- type pelagiphages, and recovered 77 nearly full- length HTVC010P- type metagenomic viral genomes from marine metagenomes. Comparative genomic and phylogenomic analyses showed that HTVC010P- type pelagiphages display genome synteny and can be clustered into two major subgroups, with subgroup I consisting of strictly lytic phages and subgroup II mostly consisting of phages with potential lysogenic life cycles. All but one member of the subgroup II contain an integrase gene. Site- specific integration of subgroup II HTVC010P- type pelagiphage was either verified experimentally or identified by in silico genomic sequence analyses, which revealed that various SAR11 tRNA genes can serve as the integration sites of HTVC010P- type pelagiphages. Moreover, HTVC010P- type pelagiphage integration was confirmed by the detection of several Global Ocean Survey (GOS) fragments that contain hybrid phage-host integration sites. Metagenomic recruitment analysis revealed that these HTVC010P- type phages were globally distributed and most lytic subgroup I members exhibited higher relative abundance. Altogether, this study significantly expands our knowledge about the genetic diversity, life strategies and ecology of HTVC010P- type pelagiphages. 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TI Diversity and biogeography of SAR11 bacteria from the Arctic Ocean SO ISME JOURNAL DT Article AB The Arctic Ocean is relatively isolated from other oceans and consists of strongly stratified water masses with distinct histories, nutrient, temperature, and salinity characteristics, therefore providing an optimal environment to investigate local adaptation. The globally distributed SAR11 bacterial group consists of multiple ecotypes that are associated with particular marine environments, yet relatively little is known about Arctic SAR11 diversity. Here, we examined SAR11 diversity using ITS analysis and metagenome-assembled genomes (MAGs). Arctic SAR11 assemblages were comprised of the S1a, S1b, S2, and S3 clades, and structured by water mass and depth. The fresher surface layer was dominated by an ecotype (S3-derived P3.2) previously associated with Arctic and brackish water. In contrast, deeper waters of Pacific origin were dominated by the P2.3 ecotype of the S2 clade, within which we identified a novel subdivision (P2.3s1) that was rare outside the Arctic Ocean. Arctic S2-derived SAR11 MAGs were restricted to high latitudes and included MAGs related to the recently defined S2b subclade, a finding consistent with bi-polar ecotypes and Arctic endemism. These results place the stratified Arctic Ocean into the SAR11 global biogeography and have identified SAR11 lineages for future investigation of adaptive evolution in the Arctic Ocean. C1 [Kraemer, Susanne; Ramachandran, Arthi; Colatriano, David; Walsh, David A.] Concordia Univ, Dept Biol, 7141 Sherbrooke St West, Sherbrooke, PQ H4B 1R6, Canada. [Lovejoy, Connie] Univ Laval, IBIS, Dept Biol, Quebec City, PQ G1K 7P4, Canada. [Lovejoy, Connie] Univ Laval, Quebec Ocean, Quebec City, PQ G1K 7P4, Canada. 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Sequenced genomes from organisms belonging to the abundant and ubiquitous SAR11 clade of marine chemoheterotrophic bacteria contain genes coding for a complete thiamin biosynthetic pathway, except for thiC, encoding the 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP) synthase, which is required for de novo synthesis of thiamin's pyrimidine moiety. Here we demonstrate that the SAR11 isolate 'Candidatus Pelagibacter ubique', strain HTCC1062, is auxotrophic for the thiamin precursor HMP, and cannot use exogenous thiamin for growth. In culture, strain HTCC1062 required 0.7 zeptomoles per cell (ca. 400 HMP molecules per cell). Measurements of dissolved HMP in the Sargasso Sea surface layer showed that HMP ranged from undetectable (detection limit: 2.4 pM) to 35.7 pM, with maximum concentrations coincident with the deep chlorophyll maximum. 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Norbeck, Angela D. Lipton, Mary S. Nicora, Carrie D. Callister, Stephen J. Smith, Richard D. Barofsky, Douglas F. Giovannoni, Stephen J. TI Proteomic analysis of stationary phase in the marine bacterium "Candidatus Pelagibacter ubique" SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB "Candidatus Pelagibacter ubique," an abundant marine alphaproteobacterium, subsists in nature at low ambient nutrient concentrations and may often be exposed to nutrient limitation, but its genome reveals no evidence of global regulatory mechanisms for adaptation to stationary phase. High-resolution capillary liquid chromatography coupled online to an LTQ mass spectrometer was used to build an accurate mass and time (AMT) tag library that enabled quantitative examination of proteomic differences between exponential- and stationary-phase "Ca. Pelagibacter ubique" cells cultivated in a seawater medium. The AMT tag library represented 65% of the predicted protein-encoding genes. "Ca. 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TI Nutrient requirements for growth of the extreme oligotroph 'Candidatus Pelagibacter ubique' HTCC1062 on a defined medium SO ISME JOURNAL DT Article AB Chemoheterotrophic marine bacteria of the SAR11 clade are Earth's most abundant organisms. Following the first cultivation of a SAR11 bacterium, 'Candidatus Pelagibacter ubique' strain HTCC1062 (Ca. P. ubique) in 2002, unusual nutritional requirements were identified for reduced sulfur compounds and glycine or serine. These requirements were linked to genome streamlining resulting from selection for efficient resource utilization in nutrient-limited ocean habitats. Here we report the first successful cultivation of Ca. P. ubique on a defined artificial seawater medium (AMS1), and an additional requirement for pyruvate or pyruvate precursors. Optimal growth was observed with the collective addition of inorganic macro-and micronutrients, vitamins, methionine, glycine and pyruvate. Methionine served as the sole sulfur source but methionine and glycine were not sufficient to support growth. Optimal cell yields were obtained when the stoichiometry between glycine and pyruvate was 1: 4, and incomplete cell division was observed in cultures starved for pyruvate. Glucose and oxaloacetate could fully replace pyruvate, but not acetate, taurine or a variety of tricarboxylic acid cycle intermediates. Moreover, both glycine betaine and serine could substitute for glycine. Interestingly, glycolate partially restored growth in the absence of glycine. We propose that this is the result of the use of glycolate, a product of phytoplankton metabolism, as both a carbon source for respiration and as a precursor to glycine. These findings are important because they provide support for the hypothesis that some micro-organisms are challenging to cultivate because of unusual nutrient requirements caused by streamlining selection and gene loss. 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Martinez-Garcia, Manuel TI Diel cycling of the cosmopolitan abundant Pelagibacter virus 37-F6: one of the most abundant viruses on earth SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB The spatiotemporal dynamics for marine viral populations has only recently been explored. However, nothing is known about temporal activities of the uncultured Pelagibacter virus vSAG 37-F6, which was discovered by single-virus genomics as potentially the most abundant marine virus. Here, we investigate the diel cycling of 37-F6 virus and the putative SAR11 host using coastal and oceanic transcriptomic and viromic time-series data from Osaka Bay and North Pacific Subtropical Gyre. Virus 37-F6 and relatives displayed diel cycling of transcriptional activities synchronized with its putative host. In both virus and host, the lowest transcription rates were observed at 14:00-15:00, coinciding roughly with maximum solar irradiance, while higher transcriptional rates were detected during the night/early morning and afternoon. Diel abundance of free viruses of 37-F6 in seawater roughly mirrored the transcriptional activities of both virus and host. In Osaka Bay, among viral relatives (genus level), virus 37-F6 specifically showed the highest ratio of transcriptional activity to virome abundance, a proxy for viral transcriptional activity relative to free viral particle abundance. This high ratio suggests high infection rate efficiencies in vSAG 37-F6 virus compared to viral relatives. Thus, time-series data revealed temporal transcript activities in one of the most abundant viruses in Earth. C1 [Martinez-Hernandez, Francisco; Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. [Luo, Elaine; DeLong, Edward F.] Univ Hawaii Manoa, Daniel K Inouye Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. [Tominaga, Kento; Yoshida, Takashi] Kyoto Univ, Grad Sch Agr, Sakyo Ku, Kyoto 6068502, Japan. [Ogata, Hiroyuki] Kyoto Univ, Inst Chem Res, Uji 6110011, Japan. RP Martinez-Garcia, M (corresponding author), Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. 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Giovannoni, Stephen J. Suffridge, Christopher P. TI Whole cell affinity for 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP) in the marine bacterium Candidatus Pelagibacter st. HTCC7211 explains marine dissolved HMP concentrations SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB Vitamin B1 is a universally required coenzyme in carbon metabolism. However, most marine microorganisms lack the complete biosynthetic pathway for this compound and must acquire thiamin, or precursor molecules, from the dissolved pool. The most common version of Vitamin B1 auxotrophy is for thiamin's pyrimidine precursor moiety, 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP). Frequent HMP auxotrophy in plankton and vanishingly low dissolved concentrations (approximately 0.1-50 pM) suggest that high-affinity HMP uptake systems are responsible for maintaining low ambient HMP concentrations. 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Davie-Martin, Cleo L. Giovannoni, Stephen J. Halsey, Kimberly H. TI Pelagibacter metabolism of diatom-derived volatile organic compounds imposes an energetic tax on photosynthetic carbon fixation SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Volatile organic compounds (VOCs) produced by phytoplankton are molecules with high vapor pressures that can diffuse across cell membranes into the environment, where they become public goods. VOCs likely comprise a significant component of the marine dissolved organic carbon (DOC) pool utilized by microorganisms, but they are often overlooked as growth substrates because their diffusivity imposes analytical challenges. The roles of VOCs in the growth of the photoautotrophic diatom Thalassiosira pseudonana and heterotrophic bacterium Pelagibacter sp. HTCC1062 (SAR11) were examined using co-cultures and proton-transfer reaction time-of-flight mass spectrometry. VOCs at 82 m/z values were produced in the cultures, and the concentrations of 9 of these m/z values changed in co-culture relative to the diatom monoculture. Several of the m/z values were putatively identified, and their metabolism by HTCC1062 was confirmed by measuring ATP production. Diatom carbon fixation rates in co-culture with HTCC1062 were 20.3% higher than the diatom monoculture. Removal of VOCs from the T. pseudonana monoculture using a hydrocarbon trap caused a similar increase in carbon fixation (18.1%). These results show that a wide range of VOCs are cycled in the environment, and the flux of VOCs from phytoplankton to bacterioplankton imposes a large and unexpected tax on phytoplankton photosynthesis. C1 [Moore, Eric R.; Davie-Martin, Cleo L.; Giovannoni, Stephen J.; Halsey, Kimberly H.] Oregon State Univ Corvallis, Dept Microbiol, 354 Nash Hall, Corvallis, OR 97331 USA. [Davie-Martin, Cleo L.] Univ Copenhagen, Terr Ecol Sect, Dept Biol, DK-2100 Copenhagen O, Denmark. 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We propose a conceptual framework in which physical mixing of the water column periodically resets SAR11 communities to a high diversity state and the seasonal evolution of phosphate limitation competitively excludes deeper-dwelling ecotypes to promote low diversity states dominated (>80%) by SAR11 Ia. A partial least squares (PLS) regression model was developed that could reliably predict sequence abundances of SAR11 ecotypes (Q(2) = 0.70) from measured environmental variables, of which mixed layer depth was quantitatively the most important. Comparison of clade-level SAR11 rRNA: rDNA signals with leucine incorporation enabled us to partially validate the use of these ratios as an in-situ activity measure. However, temporal trends in the activity of SAR11 ecotypes and their relationship to environmental variables were unclear. 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Cameron Boyd, Alex Huggett, Megan J. Grote, Jana Carini, Paul Yoder, Ryan J. Robbertse, Barbara Spatafora, Joseph W. Rappe, Michael S. Giovannoni, Stephen J. TI Phylogenomic evidence for a common ancestor of mitochondria and the SAR11 clade SO SCIENTIFIC REPORTS DT Article AB Mitochondria share a common ancestor with the Alphaproteobacteria, but determining their precise origins is challenging due to inherent difficulties in phylogenetically reconstructing ancient evolutionary events. Nonetheless, phylogenetic accuracy improves with more refined tools and expanded taxon sampling. We investigated mitochondrial origins with the benefit of new, deeply branching genome sequences from the ancient and prolific SAR11 clade of Alphaproteobacteria and publicly available alphaproteobacterial and mitochondrial genome sequences. Using the automated phylogenomic pipeline Hal, we systematically studied the effect of taxon sampling and missing data to accommodate small mitochondrial genomes. The evidence supports a common origin of mitochondria and SAR11 as a sister group to the Rickettsiales. The simplest explanation of these data is that mitochondria evolved from a planktonic marine alphaproteobacterial lineage that participated in multiple inter-specific cell colonization events, in some cases yielding parasitic relationships, but in at least one case producing a symbiosis that characterizes modern eukaryotic life. C1 [Thrash, J. Cameron; Carini, Paul; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Boyd, Alex; Yoder, Ryan J.; Robbertse, Barbara; Spatafora, Joseph W.] Oregon State Univ, Dept Bot & Plant Pathol, Corvallis, OR 97331 USA. [Huggett, Megan J.; Grote, Jana; Rappe, Michael S.] Univ Hawaii Manoa, Hawaii Inst Marine Biol, SOEST, Kaneohe, HI 96744 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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Previous studies reported that SAR11 bacteria are very small and potentially have few ribosomes, indicating that SAR11 bacteria could have low metabolic activities and could play a smaller role in the flux of dissolved organic matter than suggested by their abundance. To determine the ecological activity of SAR11 bacteria, we used a combination of microautoradiography and fluorescence in situ hybridization (Micro-FISH) to measure assimilation of (3)H-amino acids and [(35)S]dimethylsulfoniopropionate (DMSP) by SAR11 bacteria in the coastal North Atlantic Ocean and the Sargasso Sea. We found that SAR11 bacteria were often abundant in surface waters, accounting for 25% of all prokaryotes on average. SAR11 bacteria were typically as large as, if not larger than, other prokaryotes. Additionally, more than half of SAR11 bacteria assimilated dissolved amino acids and DMSP, whereas about 40% of other prokaryotes assimilated these compounds. 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SAR11 are highly abundant, cosmopolitan aquatic Alphaproteobacteria with streamlined, A+T-biased genomes. A dominant view holds that SAR11 are monophyletic and related to both Rickettsiales and the ancestor of mitochondria. Other studies dispute this, finding evidence of a polyphyletic origin of SAR11 with most strains distantly related to Rickettsiales. Although careful evolutionary modeling can reduce bias and noise in phylogenomic inference, entirely different approaches may be useful to extract robust phylogenetic signals from genomes. Here we develop simple phyloclassifiers from bioinformatically derived tRNA Class-Informative Features (CIFs), features predicted to target tRNAs for specific interactions within the tRNA interaction network. Our tRNA CIF-based model robustly and accurately classifies alphaproteobacterial genomes into one of seven undisputed monophyletic orders or families, despite great variability in tRNA gene complement sizes and base compositions. Our model robustly rejects monophyly of SAR11, classifying all but one strain as Rhizobiales with strong statistical support. Yet remarkably, conventional phylogenetic analysis of tRNAs classifies all SAR11 strains identically as Rickettsiales. We attribute this discrepancy to convergence of SAR11 and Rickettsiales tRNA base compositions. Thus, tRNA CIFs appear more robust to compositional convergence than tRNA sequences generally. Our results suggest that tRNA-CIF-based phyloclassification is robust to HGT of components of the tRNA interaction network, such as aminoacyl-tRNA synthetases. We explain why tRNAs are especially advantageous for prediction of traits governing macromolecular interactions from genomic data, and why such traits may be advantageous in the search for robust signals to address difficult problems in classification and phylogeny. Author Summary If gene products work well in the networks of foreign cells, their genes may transfer horizontally between unrelated genomes. What factors dictate the ability to integrate into foreign networks? Different RNAs and proteins must interact specifically in order to function well as a system. For example, tRNA functions are determined by the interactions they have with other macromolecules. We have developed ways to predict, from genomic data alone, how tRNAs distinguish themselves to their specific interaction partners. Here, as proof of concept, we built a robust computational model from these bioinformatic predictions in seven lineages of Alphaproteobacteria. We validated our model by classifying hundreds of diverse alphaproteobacterial taxa and tested it on eight strains of SAR11, a phylogenetically controversial group that is highly abundant in the world's oceans. We found that different strains of SAR11 are more distantly related, both to each other and to mitochondria, than widely believed. We explain conflicting results about SAR11 as an artifact of bias created by the variability in base contents of alphaproteobacterial genomes. While this bias affects tRNAs too, our classifier appears unexpectedly robust to it. More broadly, our results suggest that traits governing macromolecular interactions may be more faithfully vertically inherited than the macromolecules themselves. C1 [Amrine, Katherine C. H.; Swingley, Wesley D.; Ardell, David H.] Univ Calif Merced, Program Quantitat & Syst Biol, Merced, CA 95344 USA. RP Amrine, KCH (corresponding author), Univ Calif Merced, Program Quantitat & Syst Biol, Merced, CA 95344 USA. 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Chen, Yin TI Manganese Is Essential for PlcP Metallophosphoesterase Activity Involved in Lipid Remodeling in Abundant Marine Heterotrophic Bacteria SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB In vast areas of the ocean, microbes must adapt to the availability of scarce nutrients, and a key strategy for reducing the cellular phosphorus (P) quota is to remodel membranes by replacing phospholipids with non-P surrogate lipids. A metallo-phosphoesterase, PlcP, is essential for lipid remodeling in cosmopolitan marine bacteria of the Roseobacter (e.g., Phaeobacter sp. strain MED193) and SAR11 (e.g., Pelagibacter sp. strain HTCC7211) clades, and transcription of plcP is known to be induced by P limitation. In order to better understand PlcP-mediated lipid remodeling, we sought to characterize PlcP for its metal ion requirement and to determine its selectivity for native bacterial phospholipids. Here, we report the occurrence of a highly conserved binuclear ion center in PlcPs from MED193 and HTCC7211 and show that manganese is the preferred metal for metallophosphoesterase activity. PlcP displayed high activity towards the major bacterial phospholipids, e.g., phosphatidylglycerol but also phosphatidic acid, a key intermediate in phospholipid biosynthesis. In contrast, phosphatidylserine and phosphatidylinositol, both of which are rare lipids in bacteria, are not preferred substrates. These data suggest that PlcP undertakes a generic lipid remodeling role during the cellular response of marine bacteria to P deficiency and that manganese availability may play a key role in regulating the lipid remodeling process. IMPORTANCE Membrane lipids form the structural basis of all cells. In the marine environment, it is well established that phosphorus availability significantly affects lipid composition in cosmopolitan marine bacteria, whereby non-phosphorus-containing lipids are used to replace phospholipids in response to phosphorus stress. Central to this lipid remodeling pathway is a newly identified phospholipase C-type metallophosphoesterase (PlcP). However, little is known about how PlcP activity is regulated. Here, we determined the role of metal ions in regulating PlcP activity and compared PlcP substrate specificities in PlcP enzymes from two model marine bacteria from the marine Roseobacter clade and the SAR11 clade. Our data provide new insights into the regulation of lipid remodeling in these marine bacteria. C1 [Wei, Tao] Zhengzhou Univ Light Ind, Sch Food & Biol Engn, Zhengzhou, Henan, Peoples R China. [Wei, Tao; Quareshy, Mussa; Scanlan, David J.; Chen, Yin] Univ Warwick, Sch Life Sci, Coventry, W Midlands, England. [Zhang, Yu-Zhong] Shandong Univ, Marine Biotechnol Res Ctr, State Key Lab Microbial Technol, Jinan, Shandong, Peoples R China. [Zhang, Yu-Zhong] Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China. RP Wei, T (corresponding author), Zhengzhou Univ Light Ind, Sch Food & Biol Engn, Zhengzhou, Henan, Peoples R China.; Wei, T; Chen, Y (corresponding author), Univ Warwick, Sch Life Sci, Coventry, W Midlands, England. 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Applying MICRO-CARD-FISH, we observed that SAR11 had a large contribution to bulk abundance (37 +/- 6% of DAPI-stained cells) and to bulk bacterial heterotrophic production (BHP), as estimated from leucine incorporation (55 +/- 15% of DAPI-cells assimilating leucine) in surface waters (5 m) throughout the study period. SAR11 contributed also substantially to the assimilation of glucose, ATP, and a combination of amino acids (44 +/- 17%, 37 +/- 14%, and 43 +/- 12% of DAPI cells assimilating these compounds, respectively), organic compounds that provide either single or combined sources of C, P, and N. Temporal changes in the abundance of SAR11 cells that assimilated leucine, glucose, amino acids, and ATP revealed a pattern consistent with that of substrate-active DAPI cells, suggesting that the activity of SAR11 can explain to a large extent the variability in total cells contributing to the utilization of these compounds. Short-term nutrient enrichment experiments performed on each sampling date revealed a strong co-limitation of at least two of the three elements analyzed (C, N, P), in particular, during summer and early autumn. The in situ abundance of SAR11 cells assimilating leucine appeared to increase with P limitation as determined in the nutrient enrichment experiments (r = 0.81, p = 0.015). Our results demonstrate that SAR11 is an important component of the active bacterial community in the NW Mediterranean Sea. Our observations further indicate that the activity of the bulk bacterial community is linked to the activity of SAR11, possibly due to its adaptation to nutrient limitation. C1 [Laghdass, Melissa; Catala, Philippe; Caparros, Jocelyne; Oriol, Louise; Lebaron, Philippe; Obernosterer, Ingrid] CNRS, UMR 7621, LOM, Observ Oceanol, F-66650 Banyuls Sur Mer, France. 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The aim of the present study is to determine patterns in the biomass and productivity of Roseobacter/Rhodobacter, SAR11, and Bacteroidetes groups along the steep temperature, salinity, and organic matter gradients in the Southern Ocean by using catalyzed reporter deposition-fluorescence in situ hybridization and bromodeoxyuridine (BrdU) immunocytochemistry FISH. We found that Roseobacter/Rhodobacter, SAR11, and Bacteroidetes are prominent contributors to total bacterial biomass and production. SAR11 bacteria were the predominant lineage, but their biomass was low in the coldest regions. In contrast, the biomasses of Roseobacter/Rhodobacter and Bacteroidetes lineages were positively correlated with organic matter concentrations. The Roseobacter/Rhodobacter had the highest proportion of BrdU-positive (i.e., actively growing) cells among the three phylotypes at all stations, despite their low abundance. The relative contribution of Bacteroidetes to the total bacterial productivity (number of active cells) was negatively correlated with temperature. These results suggest that the growth and distribution patterns of Roseobacter/Rhodobacter, SAR11, and Bacteroidetes were determined by different environmental gradients (e.g., organic matter concentrations or temperature) in the Southern Ocean. C1 [Tada, Yuya; Hamasaki, Koji] Univ Tokyo, Atmosphere & Ocean Res Inst, Kashiwa, Chiba 2778564, Japan. [Makabe, Ryosuke] Hiroshima Univ, Grad Sch Biosphere Sci, Hiroshima 7398528, Japan. [Kasamatsu-Takazawa, Nobue] Tokyo Univ Marine Sci & Technol, Dept Ocean Sci, Minato Ku, Tokyo 1088477, Japan. [Taniguchi, Akito] Kinki Univ, Grad Sch Agr, Nara 6318505, Japan. RP Tada, Y (corresponding author), Univ Tokyo, Atmosphere & Ocean Res Inst, 5-1-5 Kashiwanoha, Kashiwa, Chiba 2778564, Japan. 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Ingrid TI SAR11 clade microdiversity and activity during the early spring blooms off Kerguelen Island, Southern Ocean SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB The ecology of the SAR11 clade, the most abundant bacterial group in the ocean, has been intensively studied in temperate and tropical regions, but its distribution remains largely unexplored in the Southern Ocean. Through amplicon sequencing of the 16S rRNA gene, we assessed the contribution of the SAR11 clade to bacterial community composition in the naturally iron fertilized region off Kerguelen Island. We investigated the upper 300 m at seven sites located in early spring phytoplankton blooms and at one high-nutrient low-chlorophyll site. Despite pronounced vertical patterns of the bacterioplankton assemblages, the SAR11 clade had high relative abundances at all depths and sites, averaging 40% (+/- 15%) of the total community relative abundance. 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Wang Q, 2007, APPL ENVIRON MICROB, V73, P5261, DOI 10.1128/AEM.00062-07 West NJ, 2008, ENVIRON MICROBIOL, V10, P738, DOI 10.1111/j.1462-2920.2007.01497.x West NJ, 2016, FRONT MICROBIOL, V7, DOI 10.3389/fmicb.2016.00234 NR 72 TC 7 Z9 7 PD DEC PY 2022 VL 14 IS 6 BP 907 EP 916 DI 10.1111/1758-2229.13117 EA AUG 2022 UT WOS:000846495500001 DA 2025-07-30 ER PT J AU Malmstrom, RR Cottrell, MT Elifantz, H Kirchman, DL AF Malmstrom, RR Cottrell, MT Elifantz, H Kirchman, DL TI Biomass production and assimilation of dissolved organic matter by SAR11 bacteria in the Northwest Atlantic Ocean SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Members of the SAR11 clade often dominate the composition of marine microbial communities, yet their contribution to biomass production and the flux of dissolved organic matter (DOM) is unclear. In addition, little is known about the specific components of the DOM pool utilized by SAR11 bacteria. To better understand the role of SAR11 bacteria in the flux of DOM, we examined the assimilation of leucine (a measure of biomass production), as well as free amino acids, protein, and glucose, by SAR11 bacteria in the Northwest Atlantic Ocean. We found that when SAR11 bacteria were > 25 % of total prokaryotes, they accounted for about 30 to 50 % of leucine incorporation, suggesting that SARII bacteria were major contributors to bacterial biomass production and the DOM flux. Specific growth rates of SAR11 bacteria either equaled or exceeded growth rates for the total prokaryotic community. In addition, SAR11 bacteria were typically responsible for a greater portion of amino acid assimilation (34 to 61 %) and glucose assimilation (45 to 57 %) than of protein assimilation (<= 34 %). These data suggest that SAR11 bacteria do not utilize various components of the DOM pool equally and may be more important to the flux of low-molecular-weight monomers than to that of high-molecular-weight polymers. C1 Univ Delaware, Coll Marine Studies, Lewes, DE 19958 USA. RP Univ Delaware, Coll Marine Studies, Lewes, DE 19958 USA. 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E. TI Single-cell genomics reveal low recombination frequencies in freshwater bacteria of the SAR11 clade SO GENOME BIOLOGY DT Article AB Background: The SAR11 group of Alphaproteobacteria is highly abundant in the oceans. It contains a recently diverged freshwater clade, which offers the opportunity to compare adaptations to salt-and freshwaters in a monophyletic bacterial group. However, there are no cultivated members of the freshwater SAR11 group and no genomes have been sequenced yet. Results: We isolated ten single SAR11 cells from three freshwater lakes and sequenced and assembled their genomes. A phylogeny based on 57 proteins indicates that the cells are organized into distinct microclusters. We show that the freshwater genomes have evolved primarily by the accumulation of nucleotide substitutions and that they have among the lowest ratio of recombination to mutation estimated for bacteria. In contrast, members of the marine SAR11 clade have one of the highest ratios. Additional metagenome reads from six lakes confirm low recombination frequencies for the genome overall and reveal lake-specific variations in microcluster abundances. We identify hypervariable regions with gene contents broadly similar to those in the hypervariable regions of the marine isolates, containing genes putatively coding for cell surface molecules. Conclusions: We conclude that recombination rates differ dramatically in phylogenetic sister groups of the SAR11 clade adapted to freshwater and marine ecosystems. The results suggest that the transition from marine to freshwater systems has purged diversity and resulted in reduced opportunities for recombination with divergent members of the clade. The low recombination frequencies of the LD12 clade resemble the low genetic divergence of host-restricted pathogens that have recently shifted to a new host. C1 [Zaremba-Niedzwiedzka, Katarzyna; Viklund, Johan; Zhao, Weizhou; Ast, Jennifer; Andersson, Siv G. E.] Uppsala Univ, Dept Mol Evolut, S-75124 Uppsala, Sweden. [Zaremba-Niedzwiedzka, Katarzyna; Viklund, Johan; Zhao, Weizhou; Ast, Jennifer; Andersson, Siv G. E.] Uppsala Univ, Sci Life Lab, S-75124 Uppsala, Sweden. [Sczyrba, Alexander] Univ Bielefeld, Ctr Biotechnol, D-33501 Bielefeld, Germany. [Woyke, Tanja] US DOE, Joint Genome Inst, Walnut Creek, CA 94598 USA. [McMahon, Katherina] Univ Wisconsin, Dept Civil & Environm Engn, Madison, WI 53706 USA. [McMahon, Katherina] Univ Wisconsin, Dept Bacteriol, Madison, WI 53706 USA. [Bertilsson, Stefan] Uppsala Univ, Dept Ecol & Genet, Limnol & Sci Life Lab, S-75236 Uppsala, Sweden. [Stepanauskas, Ramunas] Bigelow Lab Ocean Sci, East Boothbay, ME 04544 USA. RP Andersson, SGE (corresponding author), Uppsala Univ, Dept Mol Evolut, S-75124 Uppsala, Sweden. 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Liu, Wen-Tso TI Evolution and adaptation of SAR11 and Cyanobium in a saline Tibetan lake SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB Lake Qinghai is a unique lacustrine ecosystem located on the Tibetan Plateau and exhibits oligotrophic, alkaline, and saline conditions. Previous studies have focused on the community phylogenetic diversity of bacterioplankton in the ecosystem. This study aimed to address the ecotype diversity of bacterioplankton populations in the unique microbial habitat, using metagenomic sequencing and analysis. Phylogenetic analysis revealed two major bacterial populations: SAR11 IIIa (14% of the total) and Cyanobium (14%). Although the two populations shared high 16S rRNA gene sequence identity (> 98% identity) with their closest marine counterparts, they displayed substantial genomic divergence (<= 80% average amino acid sequence identity). Comparative genomic analysis identified conservation of carbon and energy storage metabolism (biosynthesis of polyphosphate and polyhydroxyalkanoate) gene operons in the SAR11 IIIa and a cyanate (potential nitrogen source in alkaline conditions) transporter gene operon in the Cyanobium. We further identified genetic signature of positive selection acting on an exodeoxyribonuclease gene of the SAR11 IIIa popula-tion, which is potentially associated with DNA repair responsive to strong UV radiation on the high altitude mountain. Taken together, our results revealed the ecosystem-specific gene content of the bacterioplankton populations and provided new insights into their adaptations unique to the Tibetan lake. C1 [Oh, Seungdae] Nanyang Technol Univ, Sch Civil & Environm Engn, Singapore, Singapore. [Oh, Seungdae; Liu, Wen-Tso] Univ Illinois, Dept Civil & Environm Engn, Urbana, IL USA. [Zhang, Rui] Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen, Fuijan, Peoples R China. [Zhang, Rui] Xiamen Univ, Inst Marine Microbes & Ecospheres, Xiamen, Fuijan, Peoples R China. [Wu, Qinglong L.] Chinese Acad Sci, Nanjing Inst Geog & Limnol, State Key Lab Lake Sci & Environm, Nanjing, Jiangsu, Peoples R China. RP Liu, WT (corresponding author), Univ Illinois, Dept Civil & Environm Engn, Urbana, IL USA. 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The new data expand observations about genome evolution and gene content that previously had been restricted to the SAR11 Ia subclade, providing a much broader perspective on the clade's origins, evolution, and ecology. We found small genomes throughout the clade and a very high proportion of core genome genes (48 to 56%), indicating that small genome size is probably an ancestral characteristic. In their level of core genome conservation, the members of SAR11 are outliers, the most conserved free-living bacteria known. Shared features of the clade include low GC content, high gene synteny, a large hypervariable region bounded by rRNA genes, and low numbers of paralogs. Variation among the genomes included genes for phosphorus metabolism, glycolysis, and C1 metabolism, suggesting that adaptive specialization in nutrient resource utilization is important to niche partitioning and ecotype divergence within the clade. These data provide support for the conclusion that streamlining selection for efficient cell replication in the planktonic habitat has occurred throughout the evolution and diversification of this clade. IMPORTANCE The SAR11 clade is the most abundant group of marine microorganisms worldwide, making them key players in the global carbon cycle. Growing knowledge about their biochemistry and metabolism is leading to a more mechanistic understanding of organic carbon oxidation and sequestration in the oceans. The discovery of small genomes in SAR11 provided crucial support for the theory that streamlining selection can drive genome reduction in low-nutrient environments. Study of isolates in culture revealed atypical organic nutrient requirements that can be attributed to genome reduction, such as conditional auxotrophy for glycine and its precursors, a requirement for reduced sulfur compounds, and evidence for widespread cycling of C1 compounds in marine environments. However, understanding the genetic variation and distribution of such pathways and characteristics like streamlining throughout the group has required the isolation and genome sequencing of diverse SAR11 representatives, an analysis of which we provide here. C1 [Grote, Jana; Huggett, Megan J.; Rappe, Michael S.] Univ Hawaii Manoa, Ctr Microbial Oceanog Res & Educ, SOEST, Honolulu, HI 96822 USA. [Grote, Jana; Huggett, Megan J.; Rappe, Michael S.] Univ Hawaii Manoa, Hawaii Inst Marine Biol, SOEST, Kaneohe, HI USA. [Thrash, J. Cameron; Landry, Zachary C.; Carini, Paul; Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. RP Rappé, MS (corresponding author), Univ Hawaii Manoa, Ctr Microbial Oceanog Res & Educ, SOEST, Honolulu, HI 96822 USA. 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TI Bias in assessments of marine microbial biodiversity in fosmid libraries as evaluated by pyrosequencing SO ISME JOURNAL DT Article AB On the basis of 16S rRNA gene sequencing, the SAR11 clade of marine bacteria has an almost universal distribution, being detected as abundant sequences in all marine provinces. Yet, SAR11 sequences are rarely detected in fosmid libraries, suggesting that the widespread abundance may be an artefact of PCR cloning and that SAR11 has a relatively low abundance. Here the relative abundance of SAR11 is explored in both a fosmid library and a metagenomic sequence data set from the same biological community taken from fjord surface water from Bergen, Norway. Pyrosequenced data and 16S clone data confirmed an 11-15% relative abundance of SAR11 within the community. In contrast, not a single SAR11 fosmid was identified in a pooled shotgun sequence data set of 100 fosmid clones. 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Zhang, Yu-Zhong Li, Chun-Yang TI Structure-Function Analysis Indicates that an Active-Site Water Molecule Participates in Dimethylsulfoniopropionate Cleavage by DddK SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB The osmolyte dimethylsulfoniopropionate (DMSP) is produced in petagram quantities in marine environments and has important roles in global sulfur and carbon cycling. Many marine microorganisms catabolize DMSP via DMSP lyases, generating the climate-active gas dimethyl sulfide (DMS). DMS oxidation products participate in forming cloud condensation nuclei and, thus, may influence weather and climate. SAR11 bacteria are the most abundant marine heterotrophic bacteria; many of them contain the DMSP lyase DddK, and their dddK transcripts are relatively abundant in seawater. In a recently described catalytic mechanism for DddK, Tyr64 is predicted to act as the catalytic base initiating the beta-elimination reaction of DMSP. Tyr64 was proposed to be deprotonated by coordination to the metal cofactor or its neighboring His96. To further probe this mechanism, we purified and characterized the DddK protein from Pelagibacter ubique strain HICC1062 and determined the crystal structures of wild-type DddK and its Y64A and Y122A mutants (bearing a change of Y to A at position 64 or 122, respectively), where the Y122A mutant is complexed with DMSP. The structural and mutational analyses largely support the catalytic role of Tyr64, but not the method of its deprotonation. Our data indicate that an active water molecule in the active site of DddK plays an important role in the deprotonation of Tyr64 and that this is far more likely than coordination to the metal or His96. Sequence alignment and phylogenetic analysis suggest that the proposed catalytic mechanism of DddK has universal significance. Our results provide new mechanistic insights into DddK and enrich our understanding of DMS generation by SAR11 bacteria. IMPORTANCE The climate-active gas dimethyl sulfide (DMS) plays an important role in global sulfur cycling and atmospheric chemistry. DMS is mainly produced through the bacterial cleavage of marine dimethylsulfoniopropionate (DMSP). When released into the atmosphere from the oceans, DMS can be photochemically oxidized into DMSO or sulfate aerosols, which form cloud condensation nuclei that influence the reflectivity of clouds and, thereby, global temperature. SAR11 bacteria are the most abundant marine heterotrophic bacteria, and many of them contain DMSP lyase DddK to cleave DMSP, generating DMS. In this study, based on structural analyses and mutational assays, we revealed the catalytic mechanism of DddK, which has universal significance in SAR11 bacteria. This study provides new insights into the catalytic mechanism of DddK, leading to a better understanding of how SAR11 bacteria generate DMS. 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Microbiol oxidation of DMS to DMSO represents a major sink of DMS in marine surface waters. The SAR11 clade and the marine Roseobacter clade (MRC) are the most abundant heterotrophic bacteria in the ocean surface seawater. It has been reported that trimethylamine monooxygenase (Tmm, EC 1.14.13.148) from both MRC and SAR11 bacteria likely oxidizes DMS to generate DMSO. However, the structural basis of DMS oxidation has not been explained. Here, we characterized a Tmm homolog from the SAR11 bacterium Pelagibacter sp. HTCC7211 (Tmm(7211)). Tmm(7211) exhibits DMS oxidation activity in vitro. We further solved the crystal structures of Tmm(7211) and Tmm(7211) soaked with DMS, and proposed the catalytic mechanism of Tmm(7211), which comprises a reductive half-reaction and an oxidative half-reaction. FAD and NADPH molecules are essential for the catalysis of Tmm(7211). In the reductive half-reaction, FAD is reduced by NADPH. In the oxidative half-reaction, the reduced FAD reacts with O-2 to form the C4a-(hydro)peroxyflavin. The binding of DMS may repel the nicotinamide ring of NADP(+), and make NADP(+) generate a conformational change, shutting off the substrate entrance and exposing the active C4a-(hydro)peroxyflavin to DMS to complete the oxidation of DMS. The proposed catalytic mechanism of Tmm(7211) may be widely adopted by MRC and SAR11 bacteria. This study provides important insight into the conversion of DMS into DMSO in marine bacteria, leading to a better understanding of the global sulfur cycle. C1 [Wang, Xiu-Juan; Wang, Peng; Zhang, Wei-Peng; Zhang, Yu-Zhong; Chen, Yin; Fu, Hui-Hui; Li, Chun-Yang] Ocean Univ China, Coll Marine Life Sci, Frontiers Sci Ctr Deep Ocean Multispheres & Earth, Qingdao, Peoples R China. [Wang, Xiu-Juan; Teng, Zhao-Jie; Chen, Xiu-Lan; Zhang, Yu-Zhong] Shandong Univ, Marine Biotechnol Res Ctr, State Key Lab Microbial Technol, Qingdao, Peoples R China. 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The various water masses between the subtropics/Agulhas-Benguela boundary region and the Antarctic coastal current exhibited distinct bacterioplankton communities with the highest richness in the polar frontal region, as shown by denaturing gradient gel electrophoresis of 16S rRNA gene fragments. The SAR11 clade and the Roseobacter clade-affiliated (RCA) cluster were quantified by real-time quantitative PCR. SAR11 was detected in all samples analysed from subtropical waters to the coastal current and to depths of > 1000 m. In fall and winter, this clade constituted < 3% to 48% and 4-28% of total bacterial 16S rRNA genes respectively, with highest fractions in subtropical to polar frontal regions. The RCA cluster was only present in New Zealand coastal surface waters not exceeding 17 degrees C, in the Agulhas-Benguela boundary region (visited only during the winter cruise), in subantarctic waters and in the Southern Ocean. In fall, this cluster constituted up to 36% of total bacterial 16S rRNA genes with highest fractions in the Antarctic coastal current and outnumbered the SAR11 clade at most stations in the polar frontal region and further south. In winter, the RCA cluster constituted lower proportions than the SAR11 clade and did not exceed 8% of total bacterial 16S rRNA genes. In fall, the RCA cluster exhibited significant positive correlations with latitude and ammonium concentrations and negative correlations with concentrations of nitrate, phosphate, and for near-surface samples also with chlorophyll a, biomass production of heterotrophic prokaryotes and glucose turnover rates. The findings show that the various water masses between the subtropics and the Antarctic coastal current harbour distinct bacterioplankton communities. 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Church, Matthew J. Karl, David M. Rappe, Michael S. TI Dynamics of the SAR11 bacterioplankton lineage in relation to environmental conditions in the oligotrophic North Pacific subtropical gyre SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>A quantitative PCR assay for the SAR11 clade of marine Alphaproteobacteria was applied to nucleic acids extracted from monthly depth profiles sampled over a 3-year period (2004-2007) at the open-ocean Station ALOHA (A Long-term Oligotrophic Habitat Assessment; 22 degrees 45'N, 158 degrees 00'W) in the oligotrophic North Pacific Ocean. This analysis revealed a high contribution (averaging 36% of 16S rRNA gene copies) of SAR11 to the total detected 16S rRNA gene copies over depths ranging from the surface layer to 4000 m, and revealed consistent spatial and temporal variation in the relative abundance of SAR11 16S rRNA gene copies. On average, a higher proportion of SAR11 rRNA gene copies were detected in the photic zone (< 175 m depth; mean = 38%) compared with aphotic (> 175 m depth; mean = 30%), and in the winter months compared with the summer (mean = 44% versus 33%, integrated over 175 m depth). Partial least square to latent structure projections identified environmental variables that correlate with variation in the absolute abundance of SAR11, and provided tools for developing a predictive model to explain time and depth-dependent variations in SAR11. Moreover, this information was used to hindcast temporal dynamics of the SAR11 clade between 1997 and 2006 using the existing HOT data set, which suggested that interannual variations in upper ocean SAR11 abundances were related to ocean-climate variability such as the El Nino Southern Oscillation. C1 [Eiler, Alexander; Hayakawa, Darin H.; Rappe, Michael S.] Univ Hawaii, Hawaii Inst Marine Biol, SOEST, Kaneohe, HI 96744 USA. 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Campbell, Barbara J. Kirchman, David L. TI Light-dependent growth and proteorhodopsin expression by Flavobacteria and SAR11 in experiments with Delaware coastal waters SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>Proteorhodopsin (PR)-containing bacteria are hypothesized to use both light and organic compounds as energy sources. Recent studies have found that PR is common in marine microorganisms, but the impact of light on the growth of PR-containing organisms and PR transcription in the environment remains unclear. We examined the diversity of PR genes and transcripts by PCR amplification and sequencing in Delaware coastal waters. Clone libraries of PR DNA and cDNA (from mRNA) revealed large differences between bacterial groups in expression of PR genes. We then evaluated by quantitative PCR the impact of light on growth and PR expression in PR-containing SAR11 bacteria (SAR11-PR) and a population of Flavobacteria (Flavobacteria-PR). 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Moran, Mary Ann Whitman, William B. TI Dimethylsulfoniopropionate-Dependent Demethylase (DmdA) from Pelagibacter ubique and Silicibacter pomeroyi SO JOURNAL OF BACTERIOLOGY DT Article AB The ubiquitous algal metabolite dimethylsulfoniopropionate ( DMSP) is a major source of carbon and reduced sulfur for marine bacteria. Recently, the enzyme responsible for the demethylation of DMSP, designated DmdA, was identified, and homologs were found to be common in marine bacterioplankton cells. The recombinant DmdA proteins from the cultured marine bacteria Pelagibacter ubique HTCC1062 and Silicibacter pomeroyi DSS-3 were purified with a three-step procedure using anion-exchange, hydrophobic interaction, and hydroxyapatite chromatographies. The P. ubique enzyme possessed an M-r on sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 38,500. Under nondenaturing conditions, the M-r was 68,000, suggesting that the enzyme was likely to be a dimer. The purified enzyme exhibited strict substrate specificity for DMSP, as DmdA from both S. pomeroyi and P. ubique possessed no detectable demethylase activity with glycine betaine, dimethyl glycine, methylmercaptopropionate, methionine, or dimethylsulfonioacetate. Less than 1% activity was found with dimethylsulfoniobutanoate and dimethylsulfoniopentanoate. The apparent K(m)s for DMSP were 13.2 +/- 2.0 and 5.4 +/- 2.3 mM for the P. ubique and S. pomeroyi enzymes, respectively. In cell extracts of S. pomeroyi DSS-3, the apparent K-m for DMSP was 8.6 +/- 1.2 mM, similar to that of purified recombinant DmdA. The intracellular concentration of DMSP in chemostat-grown S. pomeroyi DSS-3 was 70 mM. These results suggest that marine bacterioplankton may actively accumulate DMSP to osmotically significant concentrations that favor near-maximal rates of DMSP demethylation activity. C1 [Reisch, Chris R.; Whitman, William B.] Univ Georgia, Dept Microbiol, Athens, GA 30602 USA. 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Most studies have placed the mitochondria ancestor within the Rickettsiales order. Ten years ago, the bacterium Odyssella thessalonicensis was isolated from Acanthamoeba spp., and the 16S rDNA phylogeny placed it within the Rickettsiales. Recently, the whole genome of O. thessalonicensis has been sequenced, and 16S rDNA phylogeny and more robust and accurate phylogenomic analyses have been performed with 65 highly conserved proteins. Methodology/Principal Findings: The results suggested that the O. thessalonicensis emerged between the Rickettsiales and other Alphaproteobacteria. The mitochondrial proteins of the Reclinomonas americana have been used to locate the phylogenetic position of the mitochondrion ancestor within the Alphaproteobacteria tree. Using the K tree score method, nine mitochondrion-encoded proteins, whose phylogenies were congruent with the Alphaproteobacteria phylogenomic tree, have been selected and concatenated for Bayesian and Maximum Likelihood phylogenies. The Reclinomonas americana mitochondrion is a sister taxon to the free-living bacteria Candidatus Pelagibacter ubique, and together, they form a clade that is deeply rooted in the Rickettsiales clade. Conclusions/Significance: The Reclinomonas americana mitochondrion phylogenomic study confirmed that mitochondria emerged deeply in the Rickettsiales clade and that they are closely related to Candidatus Pelagibacter ubique. C1 [Georgiades, Kalliopi; Madoui, Mohammed-Amine; Le, Phuong; Robert, Catherine; Raoult, Didier] Univ Mediterranee, Fac Med, URMITE, CNRS IRD UMR 6236, Marseille, France. [Le, Phuong] Univ Aix Marseille 1, LATP UMR CNRS 6632, FR 3098, IFR48, Marseille, France. RP Georgiades, K (corresponding author), Univ Mediterranee, Fac Med, URMITE, CNRS IRD UMR 6236, Marseille, France. 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Analysis of known RNAs (rRNA, tRNA, riboswitches etc.) shows that structured RNAs are significantly enriched in such IGRs. To identify additional candidate structured RNAs, we examined other IGRs with similar characteristics from 'Cand. P. ubique' using comparative genomics approaches in conjunction with marine metagenomic data. Employing this strategy, we discovered four candidate structured RNAs including a new riboswitch class as well as three additional likely cis-regulatory elements that precede genes encoding ribosomal proteins S2 and S12, and the cytoplasmic protein component of the signal recognition particle. We also describe four additional potential RNA motifs with few or no examples occurring outside the metagenomic data. Conclusion: This work begins the process of identifying functional RNA motifs present in the metagenomic data and illustrates how existing completed genomes may be used to aid in this task. 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De Silva, Saumya M. Todd, Jonathan D. Dey, Mishtu TI Structural and Biochemical Insights into Dimethylsulfoniopropionate Cleavage by Cofactor-Bound DddK from the Prolific Marine Bacterium Pelagibacter SO BIOCHEMISTRY DT Article AB Enormous amounts of,the organic osmolyte drmethylsulfoniopro-pionate (DMSP) are produced in marine environments where bacterial DMSP lyases cleave it, yielding acrylate and the climate-active gas dimethyl sulfide (DMS). SAR11 bacteria are the most abundant Glade of heterotrophic bacteria in the oceans and play a key role in DMSP catabolism: An important environmental factor affecting DMS generation via DMSP lyases is the availability of metal ions because they are essential cofactors for many of these enzymes. Here we examine the structure and activity of DddK in the presence of various metal ions. We have established that DddK containing a double-stranded beta-helical, motif utilizes various divalent metal Rills as cofactors for catalytic activity. 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Zemskaya, Tamara I. Rosselli, Riccardo Coutinho, Felipe H. Zakharenko, Alexandra S. Blinov, Vadim V. Rodriguez-Valera, Francisco TI Genomes of Novel Microbial Lineages Assembled from the Sub-Ice Waters of Lake Baikal SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB We present a metagenomic study of Lake Baikal (East Siberia). Two samples obtained from the water column under the ice cover (5 and 20 m deep) in March 2016 have been deep sequenced and the reads assembled to generate metagenome-assembled genomes (MAGs) that are representative of the microbes living in this special environment. Compared with freshwater bodies studied around the world, Lake Baikal had an unusually high fraction of Verrucomicrobia. Other groups, such as Actinobacteria and Proteobacteria, were in proportions similar to those found in other lakes. The genomes (and probably cells) tended to be small, presumably reflecting the extremely oligotrophic and cold prevalent conditions. Baikal microbes are novel lineages recruiting very little from other water bodies and are distantly related to other freshwater microbes. Despite their novelty, they showed the closest relationship to genomes discovered by similar approaches from other freshwater lakes and reservoirs. Some of them were particularly similar to MAGs from the Baltic Sea, which, although it is brackish, connected to the ocean, and much more eutrophic, has similar climatological conditions. Many of the microbes contained rhodopsin genes, indicating that, in spite of the decreased light penetration allowed by the thick ice/snow cover, photoheterotrophy could be widespread in the water column, either because enough light penetrates or because the microbes are already adapted to the summer ice-less conditions. We have found a freshwater SAR11 subtype I/II representative showing striking synteny with Pelagibacter ubique strains, as well as a phage infecting the widespread freshwater bacterium Polynucleobacter. IMPORTANCE Despite the increasing number of metagenomic studies on different freshwater bodies, there is still a missing component in oligotrophic cold lakes suffering from long seasonal frozen cycles. Here, we describe microbial genomes from metagenomic assemblies that appear in the upper water column of Lake Baikal, the largest and deepest freshwater body on Earth. This lake is frozen from January to May, which generates conditions that include an inverted temperature gradient (colder up), decrease in light penetration due to ice, and, especially, snow cover, and oligotrophic conditions more similar to the open-ocean and high-altitude lakes than to other freshwater or brackish systems. As could be expected, most reconstructed genomes are novel lineages distantly related to others in cold environments, like the Baltic Sea and other freshwater lakes. 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TI Differential responses of Prochlorococcus and SAR11-dominated bacterioplankton groups to atmospheric dust inputs in the tropical Northeast Atlantic Ocean SO FEMS MICROBIOLOGY LETTERS DT Article AB The metabolic responses of indigenous dominant bacterioplankton populations to additions of dust were examined in the tropical northeast Atlantic. Subsurface seawater samples were treated with dust, added directly or indirectly as a 'leachate' after its rapid dissolution in deionized water. Samples were incubated at ambient temperature and light for up to 24 h and microbial metabolic responses were assessed by 35S-methionine (35S-Met) uptake. Prochlorococcus and low nucleic acid (LNA) cells were sorted by flow cytometry to determine their group-specific responses. Sorted cells were also phylogenetically affiliated using FISH. The high-light-adapted ecotype II dominated the Prochlorococcus group and 73 +/- 14% of LNA prokaryotes belonged to the SAR11 clade of Alphaproteobacteria. Both Prochlorococcus and LNA cells were metabolically impaired by the addition of dust (40 +/- 28% and 37 +/- 22% decrease in 35S-Met uptake compared with controls, respectively). However, LNA bacterioplankton showed minor positive responses to dust leachate additions (7 +/- 4% increase in 35S-Met uptake), while the metabolic activity of Prochlorococcus cells decreased in the presence of dust leachate by 16 +/- 11%. Thus, dust dissolution in situ appears to be more deleterious to Prochlorococcus than SAR11-dominated LNA bacterioplankton and hence could initiate a compositional shift in the indigenous bacterioplankton. C1 [Hill, Polly G.; Zubkov, Mikhail V.; Purdie, Duncan A.] Univ Southampton, Natl Oceanog Ctr, Southampton SO14 3ZH, Hants, England. RP Hill, PG (corresponding author), Univ Southampton, Natl Oceanog Ctr, European Way, Southampton SO14 3ZH, Hants, England. 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Carlson, Craig A. TI Basin-scale patterns in the abundance of SAR11 subclades, marine Actinobacteria (OM1), members of the Roseobacter clade and OCS116 in the South Atlantic SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Bacterioplankton are major biogeochemical agents responsible for mediating the flux of dissolved organic matter (DOM) and subsequent cycling of nutrients in the oceans. Most information about the composition of bacterioplankton communities has come from studies along well-defined biogeochemical gradients in the northern hemisphere. This study extends observations of spatial and temporal dynamics for SAR11, Actinobacteria and OCS116 in the North Atlantic by demonstrating distinct spatial variability in the abundance and distribution of these and other lineages across the South Atlantic gyre and in the Benguela upwelling system. We identified shifts in SAR11, Actinobacteria, OCS116, SAR86, SAR116 and members of the Roseobacter clade along basin-scale gradients in nutrients, chlorophyll and dissolved organic carbon (DOC). Distinct SAR11 subclades dominated the western and eastern regions of the gyre, and Actinobacteria, OCS116 and members of the Roseobacter lineages were most abundant at the deep chlorophyll maxima. SAR86 and SAR116 accounted for a significant fraction of coastal and open ocean communities, respectively, and members of the gamma sulfur oxidizer (GSO) clade persisted in the Benguela upwelling system. These data suggest that distinct communities are partitioned along basin-scale biogeochemical gradients, that SAR11 community structure varies across the gyre and that Actinobacteria, OCS116, and members of the Roseobacter clade are closely associated with phytoplankton in the gyre. C1 [Morris, Robert M.; Frazar, Christian D.] Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. 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Lipski, Andre Iversen, Morten H. Mollenhauer, Gesine TI Microbial communities in the nepheloid layers and hypoxic zones of the Canary Current upwelling system SO MICROBIOLOGYOPEN DT Article AB Eastern boundary upwelling systems (EBUSs) are among the most productive marine environments in the world. The Canary Current upwelling system off the coast of Mauritania and Morocco is the second most productive of the four EBUS, where nutrient-rich waters fuel perennial phytoplankton blooms, evident by high chlorophyll a concentrations off Cape Blanc, Mauritania. High primary production leads to eutrophic waters in the surface layers, whereas sinking phytoplankton debris and horizontally dispersed particles form nepheloid layers (NLs) and hypoxic waters at depth. We used Catalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) in combination with fatty acid (measured as methyl ester; FAME) profiles to investigate the bacterial and archaeal community composition along transects from neritic to pelagic waters within the "giant Cape Blanc filament" in two consecutive years (2010 and 2011), and to evaluate the usage of FAME data for microbial community studies. We also report the first fatty acid profile of Pelagibacterales strain HTCC7211 which was used as a reference profile for the SAR11 clade. Unexpectedly, the reference profile contained low concentrations of long chain fatty acids 18:1 cis11, 18:1 cis11 11methyl, and 19:0 cyclo11-12 fatty acids, the main compounds in other Alphaproteobacteria. Members of the free-living SAR11 clade were found at increased relative abundance in the hypoxic waters in both years. In contrast, the depth profiles of Gammaproteobacteria (including Alteromonas and Pseudoalteromonas), Bacteroidetes, Roseobacter, and Synechococcus showed high abundances of these groups in layers where particle abundance was high, suggesting that particle attachment or association is an important mechanisms of dispersal for these groups. Collectively, our results highlight the influence of NLs, horizontal particle transport, and low oxygen on the structure and dispersal of microbial communities in upwelling systems. C1 [Thiele, Stefan] Max Planck Inst Marine Microbiol, Bremen, Germany. [Basse, Andreas; Iversen, Morten H.; Mollenhauer, Gesine] Alfred Wegener Inst Polar & Marine Res, Handelshafen 12, D-27570 Bremerhaven, Germany. [Becker, Jamie W.] Haverford Coll, Dept Biol, Haverford, PA 19041 USA. [Lipski, Andre] Rheinische Friedrich Wilhelms Univ Bonn, Dept Food Microbiol & Hyg, Bonn, Germany. [Basse, Andreas; Iversen, Morten H.; Mollenhauer, Gesine] MARUM, Bremen, Germany. 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Its mild salinity gradient provides a unique environment for the coexistence of freshwater and marine bacteria. This study employs catalysed reporter deposition-fluorescence in situ hybridisation (CARD-FISH) to investigate seasonal dynamics of marine (SAR11 clade I/II) and freshwater (SAR11 clade IIIb (LD12), Limnohabitans clades B, C, D, Burkholderiales (former Betaproteobacteria)) bacterial groups. Samples were collected from below the ice in February, and then monthly from April to October 2011 from three stations with different salinity. The abundance of SAR11 clade I/II strongly positively correlated with salinity, with the highest abundance in autumn at time of inflows of saline waters from the Baltic Sea. Two groups (spring-summer and autumn-winter) were distinguished within this clade, based on a scatter chart between SAR11-I/II abundance and salinity. Salinity explained 69% of the variability of the spring-summer group and 77% of the autumn-winter group. This suggests that the presence of marine SAR11-I/II in the Vistula Lagoon was caused by passive inflow with waters from the open Baltic Sea, and this environment is not suitable for its regular existence. The abundance of the freshwater SAR11 clade IIIb was similar to that of SAR11-I/II. However, it depended on temperature, organic and inorganic phosphorous, and not on salinity, suggesting that SAR11-IIIb found a regular niche in the eutrophic Vistula Lagoon. Burkholderiales positively correlated with temperature, chlorophyll-a, organic and inorganic phosphorous, and heterotrophic nanoflagellates, suggesting that their role in the Vistula Lagoon may be similar to that in lakes. Interestingly, Burkholderiales had the highest relative abundance under ice in February. Finally, the abundance of Limnohabitans clades B, C, D positively depended on temperature, chlorophyll-a and negatively on salinity. 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Orellana, Luis H. Sidhu, Chandni Klip, Helena C. L. Meunier, Cedric L. Boersma, Maarten Wiltshire, Karen H. Amann, Rudolf Fuchs, Bernhard M. TI In situ cell division and mortality rates of SAR11, SAR86, Bacteroidetes, and Aurantivirga during phytoplankton blooms reveal differences in population controls SO MSYSTEMS DT Article AB Net growth of microbial populations, that is, changes in abundances over time, can be studied using 16S rRNA fluorescence in situ hybridization (FISH). However, this approach does not differentiate between mortality and cell division rates. We used FISH-based image cytometry in combination with dilution culture experiments to study net growth, cell division, and mortality rates of four bacterial taxa over two distinct phytoplankton blooms: the oligotrophs SAR11 and SAR86, and the copiotrophic phylum Bacteroidetes, and its genus Aurantivirga. Cell volumes, ribosome content, and frequency of dividing cells (FDC) co-varied over time. Among the three, FDC was the most suitable predictor to calculate cell division rates for the selected taxa. The FDC-derived cell division rates for SAR86 of up to 0.8/day and Aurantivirga of up to 1.9/day differed, as expected for oligotrophs and copiotrophs. Surprisingly, SAR11 also reached high cell division rates of up to 1.9/day, even before the onset of phytoplankton blooms. For all four taxonomic groups, the abundance-derived net growth (-0.6 to 0.5/day) was about an order of magnitude lower than the cell division rates. Consequently, mortality rates were comparably high to cell division rates, indicating that about 90% of bacterial production is recycled without apparent time lag within 1 day. Our study shows that determining taxon-specific cell division rates complements omics-based tools and provides unprecedented clues on individual bacterial growth strategies including bottom-up and top-down controls. IMPORTANCE The growth of a microbial population is often calculated from their numerical abundance over time. However, this does not take cell division and mortality rates into account, which are important for deriving ecological processes like bottom-up and top-down control. In this study, we determined growth by numerical abundance and calibrated microscopy-based methods to determine the frequency of dividing cells and subsequently calculate taxon-specific cell division rates in situ. The cell division and mortality rates of two oligotrophic (SAR11 and SAR86) and two copiotrophic (Bacteroidetes and Aurantivirga) taxa during two spring phytoplankton blooms showed a tight coupling for all four taxa throughout the blooms without any temporal offset. Unexpectedly, SAR11 showed high cell division rates days before the bloom while cell abundances remained constant, which is indicative of strong top-down control. Microscopy remains the method of choice to understand ecological processes like top-down and bottom-up control on a cellular level. C1 [Bruewer, Jan D.; Orellana, Luis H.; Sidhu, Chandni; Amann, Rudolf; Fuchs, Bernhard M.] Max Planck Inst Marine Microbiol, Bremen, Germany. [Klip, Helena C. L.; Meunier, Cedric L.; Boersma, Maarten; Wiltshire, Karen H.] Alfred Wegener Inst, Biol Anstalt Helgoland, Helmholtz Zentrum Polar & Meeresforsch, Helgoland, Germany. [Boersma, Maarten] Univ Bremen, Bremen, Germany. [Wiltshire, Karen H.] Alfred Wegener Inst, Helmholtz Zentrum Polar & Meeresforsch, Wattenmeerstn, Bremerhaven, Germany. RP Fuchs, BM (corresponding author), Max Planck Inst Marine Microbiol, Bremen, Germany. 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HTCC7211 SO BIOTECHNOLOGY LETTERS DT Article AB To characterize a novel thermophilic beta-carotene 15,15 '-monooxygenase BCMO7211 isolated from the marine bacterium Candidatus Pelagibacter sp. HTCC7211. BCMO7211 was functionally overexpressed in Escherichia coli and purified to homogeneity by Ni-NTA affinity chromatography and Superdex-200 gel filtration chromatography. Labeling experiments with (H2O)-O-18 demonstrated that the oxygen atom in the terminal aldehyde group of the produced retinal molecules was provided from both molecular oxygen and water, indicating that BCMO7211 is the first characterized bacterial beta-carotene 15,15 '-monooxygenase. BCMO7211 exhibited broad carotenoid substrate specificity toward alpha-carotene, beta-cryptoxanthin, beta-carotene, zeaxanthin, and lutein. The optimum temperature, pH, and concentrations of the substrate and enzyme for retinal production were 60 degrees C, 9.0, 500 mg beta-carotene/L, and 2.5 U/ml, respectively. 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This paring down the bacterium's genome appears to be evolution's way of conserving genetic energy for an abundant organism in the face of limited resources. NR 0 TC 0 Z9 0 PD DEC PY 2005 VL 293 IS 6 BP 26 EP 28 DI 10.1038/scientificamerican1205-26 UT WOS:000233750200019 DA 2025-07-30 ER PT J AU Mohapatra, M Behera, P Kim, JY Rastogi, G AF Mohapatra, Madhusmita Behera, Pratiksha Kim, Ji Yoon Rastogi, Gurdeep TI Seasonal and spatial dynamics of bacterioplankton communities in a brackish water coastal lagoon SO SCIENCE OF THE TOTAL ENVIRONMENT DT Article AB Coastal ecosystems, one of the most productive ecosystems, are subjected to natural and anthropogenic stresses. Coastal bacterioplankton communities are highly dynamic due to spatiotemporal heterogeneity in the environmental parameters. 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This study has provided baseline information on the bacterioplankton communities and their environmental drivers within an anthropogenically impacted cyclone prone coastal lagoon which would be useful in assessing the impact of multiple stressors on this vulnerable ecosystem. (C) 2019 Elsevier B.V. All rights reserved. C1 [Mohapatra, Madhusmita; Behera, Pratiksha; Rastogi, Gurdeep] Chilika Dev Author, Wetland Res & Training Ctr, Balugaon 752030, Odisha, India. [Mohapatra, Madhusmita] KIIT Univ, Sch Biotechnol, Bhubaneswar 751024, Odisha, India. [Kim, Ji Yoon] Natl Inst Environm Studies, Ctr Climate Change Adaptat, Tsukuba, Ibaraki 3058506, Japan. RP Rastogi, G (corresponding author), Chilika Dev Author, Wetland Res & Training Ctr, Balugaon 752030, Odisha, India. 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Lang, Andrew S. TI Effects of Vertical Water Mass Segregation on Bacterial Community Structure in the Beaufort Sea SO MICROORGANISMS DT Article AB The Arctic Ocean is one of the least well-studied marine microbial ecosystems. Its low-temperature and low-salinity conditions are expected to result in distinct bacterial communities, in comparison to lower latitude oceans. However, this is an ocean currently in flux, with climate change exerting pronounced effects on sea-ice coverage and freshwater inputs. How such changes will affect this ecosystem are poorly constrained. In this study, we characterized the bacterial community compositions at different depths in both coastal, freshwater-influenced, and pelagic, sea-ice-covered locations in the Beaufort Sea in the western Canadian Arctic Ocean. The environmental factors controlling the bacterial community composition and diversity were investigated. 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Kirchman, David L. TI Transcriptional Control in Marine Copiotrophic and Oligotrophic Bacteria with Streamlined Genomes SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Bacteria often respond to environmental stimuli using transcriptional control, but this may not be the case for marine bacteria such as "Candidatus Pelagibacter ubique," a cultivated representative of the SAR11 clade, the most abundant organism in the ocean. This bacterium has a small, streamlined genome and an unusually low number of transcriptional regulators, suggesting that transcriptional control is low in Pelagibacter and limits its response to environmental conditions. Transcriptome sequencing during batch culture growth revealed that only 0.1% of protein-encoding genes appear to be under transcriptional control in Pelagibacter and in another oligotroph (SAR92) whereas >10% of genes were under transcriptional control in the copiotrophs Polaribacter sp. strain MED152 and Ruegeria pomeroyi. 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Sanchez, Olga Gonzalez, Jose M. Pinhassi, Jarone Massana, Ramon Pernthaler, Jakob Pedros-Alio, Carlos Gasol, Josep M. TI Seasonality in bacterial diversity in north-west Mediterranean coastal waters:: assessment through clone libraries, fingerprinting and FISH SO FEMS MICROBIOLOGY ECOLOGY DT Article AB We combined denaturing gradient gel electrophoresis (DGGE), catalysed reporter deposition-FISH (CARD-FISH) and clone libraries to investigate the seasonality of the bacterial assemblage composition in north-west Mediterranean coastal waters. DGGE analysis indicated that bacterial diversity changed gradually throughout the year, although with a clear distinction of the summer period. Alphaproteobacteria were the dominant group on an annual basis [29% of the DAPI (4',6-diamidino-2-phenylindole) counts by CARD-FISH, and 70% of the bacterial clones]. 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In the tropical waters, this is a permanent feature, but in the Mediterranean and other temperate waters, the DCM is a seasonal phenomenon. The metagenome from a single sample of a mature Mediterranean DCM community has been 454 pyrosequenced both directly and after cloning in fosmids. This study is the first to be carried out at this sequencing depth (ca. 600 Mb combining direct and fosmid sequencing) at any DCM. Our results indicate a microbial community massively dominated by the high-light-adapted Prochlorococcus marinus subsp. pastoris, Synechococcus sp., and the heterotroph Candidatus Pelagibacter. The sequences retrieved were remarkably similar to the existing genome of P. marinus subsp. pastoris with a nucleotide identity over 98%. Besides, we found a large number of cyanophages that could prey on this microbe, although sequence conservation was much lower. 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Thrash, J. Cameron TI Microbial ecology of northern Gulf of Mexico estuarine waters SO MSYSTEMS DT Article AB Estuarine and coastal ecosystems are of high economic and ecological importance, owing to their diverse communities and the disproportionate role they play in carbon cycling, particularly in carbon sequestration. Organisms inhabiting these environments must overcome strong natural fluctuations in salinity, nutrients, and turbidity, as well as numerous climate change-induced disturbances such as land loss, sea level rise, and, in some locations, increasingly severe tropical cyclones that threaten to disrupt future ecosystem health. The northern Gulf of Mexico (nGoM) along the Louisiana coast contains dozens of estuaries, including the Mississippi-Atchafalaya River outflow, which dramatically influence the region due to their vast upstream watershed. Nevertheless, the microbiology of these estuaries and surrounding coastal environments has received little attention. To improve our understanding of microbial ecology in the understudied coastal nGoM, we conducted a 16S rRNA gene amplicon survey at eight sites and multiple time points along the Louisiana coast and one inland swamp spanning freshwater to high brackish salinities, totaling 47 duplicated Sterivex (0.2-2.7 mu m) and prefilter (>2.7 mu m) samples. We cataloged over 13,000 Amplicon Sequence ariants (ASVs) from common freshwater and marine clades such as SAR11 (Alphaproteobacteria), Synechococcus (Cyanobacteria), and acI and Candidatus Actinomarina (Actinobacteria). We observed correlations with freshwater or marine habitats in many organisms and characterized a group of taxa with specialized distributions across brackish water sites, supporting the hypothesis of an endogenous brackish-water community. Additionally, we observed brackish-water associations for several aquatic clades typically considered marine or freshwater taxa, such as SAR11 subclade II, SAR324, and the acI Actinobacteria. The data presented here expand the geographic coverage of microbial ecology in estuarine communities, help delineate the native and transitory members of these environments, and provide critical aquatic microbiological baseline data for coastal and estuarine sites in the nGoM. C1 [Henson, Michael W.] Northern Univ, Dept Biol Sci, De Kalb, IL 60115 USA. [Thrash, J. Cameron] Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA. RP Henson, MW (corresponding author), Northern Univ, Dept Biol Sci, De Kalb, IL 60115 USA.; Thrash, JC (corresponding author), Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA. 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Cottrel, Matthew T. DiTullio, Giacomo R. TI Shaping of bacterial community composition and diversity by phytoplankton and salinity in the Delaware Estuary, USA SO AQUATIC MICROBIAL ECOLOGY DT Article AB Our understanding of the impact of phytoplankton on bacterial communities is largely based on studies showing that a few bacteria have interactions with the phytoplankton class, which is often diatoms, dominating phytoplankton communities. To determine the effect of the complete phytoplankton community on the entire bacterial community, we used tag pyro sequences of the 16S rRNA gene and phytoplankton pigments measured by high performance liquid chromatography along with Chemtax analyses to examine bacterial and phytoplankton communities along the salinity gradient of the Delaware Estuary, USA, in August and November of 3 years (2011-2013). Salinity had a large effect on the composition, taxon richness, and evenness of bacterial communities in the estuary, but so too did the composition and biomass of the phytoplankton community. Phytoplankton classes had a larger effect in shaping the composition of bacterial communities than did total chlorophyll a. Although diatoms and cryptophytes dominated the phytoplankton communities in both August and November, less common phytoplankton classes, such as dinoflagellates, haptophytes, and prasinophytes, had more significant relationships with the entire bacterial community and with individual bacterial taxa. In contrast, the 2 most abundant bacterial subclades in the estuary, SAR11 IIIa and SAR 11 IIIb, had few significant relationships with chlorophyll a or with phytoplankton classes. These data on bacterial and phytoplankton community composition help to explain the weak coupling between bacteria and phytoplankton communities often observed in estuarine and other aquatic systems. 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Cottrell, Matthew T. Kirchman, David L. TI Growth rates and rRNA content of four marine bacteria in pure cultures and in the Delaware estuary SO ISME JOURNAL DT Article AB Interpretation of 16S ribosomal RNA (rRNA) to 16S rRNA gene ratios (rRNA: rDNA) is based on a limited number of studies with rapidly growing copiotrophic bacteria. The most abundant bacteria in the ocean are oligotrophs, which probably grow more slowly than those bacteria whose rRNA: rDNA versus growth rate relationships are known. To examine whether rRNA: rDNA varies differently in oligotrophic marine bacteria than in copiotrophic bacteria, we used quantitative PCR and reverse transcriptase quantitative PCR to measure rRNA: rDNA in two marine copiotrophs and in two marine oligotrophs, including Candidatus Pelagibacter ubique HTCC1062, a coastal isolate of SAR11, the most abundant bacterial clade in the ocean. 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Waidner, Lisa A. Cottrell, Matthew T. Kirchman, David L. TI Abundant proteorhodopsin genes in the north atlantic ocean SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Proteorhodopsin (PR) is a light-driven proton pump that has been found in a variety of marine bacteria, including Pelagibacter ubique, a member of the ubiquitous SAR11 clade. The goals of this study were to explore the diversity of PR genes and to estimate their abundance in the North Atlantic Ocean using quantitative polymerase chain reaction (QPCR). We found that PR genes in the western portion of the Sargasso Sea could be grouped into 27 clusters, but five clades had the most sequences. Sets of specific QPCR primers were designed to examine the abundance of PR genes in the following four of the five clades: SAR11 (P. ubique and other SAR11 Alphaproteobacteria), BACRED17H8 (Alphaproteobacteria), HOT2C01 (Alphaproteobacteria) and an uncultured subgroup of the Flavobacteria. 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Wang, Da-Zhi TI Phosphoproteomics reveals essential regulatory roles of phosphorylation in marine oligotrophic bacteria SO MARINE LIFE SCIENCE & TECHNOLOGY DT Article; Early Access AB Oligotrophic bacteria with reduced genomes have relatively few transcriptional regulators and are thought to rely more than other bacteria on post-transcriptional regulation to respond to environmental stimuli. SAR11 bacteria are the most abundant group of heterotrophic bacteria in marine planktonic systems and are a model for understanding genome reduction in other free-living microorganisms. Here, we report a comprehensive, quantitative protein phosphorylation profile for SAR11 strain HTCC1062 grown under various environmentally relevant conditions, including light/dark cycles, temperature differences, and nutrient limitations, to investigate phosphorylation dynamics in this streamlined organism. Nearly half of proteins encoded by the genome were detected in phosphorylated forms under at least one condition. 1014 Ser/Thr/Tyr phosphorylation sites were observed in 1576 phosphopeptides from 555 phosphoproteins. Protein phosphorylation was concentrated in proteins for functions associated with nutrient acquisition and growth, such as ABC transporters, RNA polymerase, and ribosomal proteins. Prominent patterns in protein phosphorylation were detected across a range of culture conditions. In these cells, which previously have been shown to continuously express nearly their entire proteome, protein phosphorylation was more dynamic than protein abundance, supporting the hypothesis that post-transcriptional regulation by protein phosphorylation might play a large role in modulating protein activity. Our findings support a regulatory model characterized by minimal variation in protein expression but extensive protein phosphorylation. This model diverges from bacterial regulatory paradigms reliant on transcriptional control, and may be relevant to understanding other abundant heterotrophs with reduced genomes. C1 [Zhang, Yu; Xie, Zhang-Xian; Bai, Zhuo-An; Wang, Ming-Hua; Wang, Da-Zhi] Xiamen Univ, Coll Environm & Ecol, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China. [He, Yao-Hui] Univ South China, Hengyang Med Sch, MOE Key Lab Rare Pediat Dis, Hengyang 421001, Peoples R China. [Xie, Zhang-Xian] Quanzhou Normal Univ, Sch Resources & Environm Sci, Key Lab Rural Environm Remediat & Waste Recycling, Quanzhou 362000, Peoples R China. [Hu, Guo-Sheng] Xiamen Univ, Sch Pharmaceut Sci, Fujian Prov Key Lab Innovat Drug Target Res, Xiamen 361102, Peoples R China. [Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. RP Wang, DZ (corresponding author), Xiamen Univ, Coll Environm & Ecol, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China.; Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. 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HGT is widespread in prokaryotes, contributing to the genomic repertoire of prokaryotic organisms, and is particularly apparent in Rickettsiales genomes. Gene gains from both distantly and closely related organisms play crucial roles in the evolution of bacterial genomes. In this work, we focus on genes transferred from distantly related species into Rickettsiales species. Results: We developed an automated approach for the detection of HGT from other organisms (excluding alphaproteobacteria) into Rickettsiales genomes. Our systematic approach consisted of several specialized features including the application of a parsimony method for inferring phyletic patterns followed by blast filter, automated phylogenetic reconstruction and the application of patterns for HGT detection. We identified 42 instances of HGT in 31 complete Rickettsiales genomes, of which 38 were previously unidentified instances of HGT from Anaplasma, Wolbachia, Candidatus Pelagibacter ubique and Rickettsia genomes. 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Santoro, Alyson E. TI Exaggerated trans-membrane charge of ammonium transporters in nutrient-poor marine environments SO OPEN BIOLOGY DT Article AB Transporter proteins are a vital interface between cells and their environment. In nutrient-limited environments, microbes with transporters that are effective at bringing substrates into their cells will gain a competitive advantage over variants with reduced transport function. Microbial ammonium transporters (Amt) bring ammonium into the cytoplasm from the surrounding periplasm space, but diagnosing Amt adaptations to low nutrient environments solely from sequence data has been elusive. Here, we report altered Amt sequence amino acid distribution from deep marine samples compared to variants sampled from shallow water in two important microbial lineages of the marine water column community-Marine Group I Archaea (Thermoproteota) and the uncultivated gammaproteobacterial lineage SAR86. 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The data suggest that a significant fraction of viruses in the mid-euphotic zone of the subtropical gyres may be cyanophages and patterns in their abundance are largely determined by Prochlorococcus dynamics in response to water column stability. This high-resolution, decadal survey of virioplankton abundance provides insight into the possible controls of virioplankton dynamics in the open ocean. The ISME Journal (2012) 6, 273-284; doi: 10.1038/ismej.2011.101; published online 11 August 2011 C1 [Carlson, Craig A.] Univ Calif Santa Barbara, Dept Ecol Evolut & Marine Biol, Santa Barbara, CA 93106 USA. [Parsons, Rachel J.; Lomas, Michael W.] Bermuda Inst Ocean Sci, St Georges GE01, Bermuda. [Breitbart, Mya] Univ S Florida, Coll Marine Sci, St Petersburg, FL 33701 USA. RP Carlson, CA (corresponding author), Univ Calif Santa Barbara, Dept Ecol Evolut & Marine Biol, Santa Barbara, CA 93106 USA. 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A primary method of determining their diversity and distribution is through sequencing of 16S ribosomal RNA genes from environmental samples. However, the perceived community composition may vary significantly based on differences in methodology, including choice of 16S variable region(s). This study investigated the influence of 16S variable region selection (V4-V5 or V6-V8) on perceived community composition and diversity for bacteria, Archaea and chloroplasts by tag-Illumina sequencing. We used 24 samples from the photic zone of the Scotian Shelf, northwest Atlantic, collected during a spring phytoplankton bloom. Taxonomic assignment and community composition varied greatly depending on the choice of variable regions while observed patterns of beta diversity were reproducible between variable regions. V4-V5 was considered the preferred variable region for future studies based on its superior recognition of Archaea, which has received little attention in bloom dynamics. 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(2) Methods: Marine prokaryotic diversity was investigated using high-throughput sequencing of the 16S rRNA gene during the bloom declines of two dinoflagellates detected in the summer of 2019 along the northern and southern Tunisian coasts (South Mediterranean Sea). The species Gymnodinium impudicum (Carthage, Tunis Gulf) and Alexandrium minutum (Sfax, Gabes Gulf) were identified using microscopy and molecular methods and were related to physicochemical factors and prokaryotic compositions. (3) Results: The abundance of G. impudicum decreased over time with decreasing phosphate concentrations. During the G. impudicum bloom decay, prokaryotes were predominated by the archaeal MGII group (Thalassarchaeaceae), Pelagibacterales (SAR11), Rhodobacterales, and Flavobacteriales. At Sfax, the abundance of A. minutum declined with decreasing phosphate concentrations and increasing pH. 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Oligonucleotide probes were designed to enumerate some bacterial groups found in the clone library such as the Arctic96B-16 cluster, Roseobacter RCA cluster, AGG58 cluster, and Polaribacter clade. FISH analyses revealed that each of these groups typically accounted for 3 to 10% of prokaryotes, while well known cosmopolitan groups like the SAR86 and SAR11 clades made up 8 and 25% of the prokaryotic communities, respectively. Overall, > 60% of total prokaryotes belonged to 8 specific bacterial groups, of which the SAR11 clade was the most diverse (< 13% 16S rRNA sequence difference). Together the Arctic96B-16, Roseobacter RCA, Polaribacter, and SARI 1 clades also accounted for 25 to 82% of biomass production at 4 locations, as determined by a combination of FISH and microautoradiography of H-3-leucine assimilation. These results indicate that community composition and biomass production in the western Arctic Ocean are dominated by a few bacterial groups. 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Ertan, Haluk Ting, Lily Cavicchioli, Ricardo TI Carbon and nitrogen substrate utilization in the marine bacterium Sphingopyxis alaskensis strain RB2256 SO ISME JOURNAL DT Article AB Sphingopyxis alaskensis is a marine member of the Alphaproteobacteria that is adapted to heterotrophic growth under nutrient-depleted (oligotrophic) conditions. S. alaskensis strain RB2256 is an ultramicrobacterium (cell volume <0.1 mu m(3)), and has a genome size larger than that of the ultramicrobacterium 'Candidatus Pelagibacter ubique' HTCC1062 (SAR11 clade of Alphaproteobacteria): 3.35 versus 1.31 Mbp. In this study, we investigate the carbon and nitrogen metabolism of strain RB2256 using an integrated approach that combines growth and enzyme assays, proteomics and genome analysis. S. alaskensis is able to use specific amino acids and putrescine as a sole carbon and nitrogen source, and higher energy-yielding substrates such as glucose and trehalose as carbon sources. Alanine, in particular, emerges as a very important substrate in S. alaskensis metabolism. In an oligotrophic environment where competition for nutrients is intense, our data support a simplified metabolism for S. alaskensis in which the fate of certain substrates is constrained, especially at the intersections of central carbon and nitrogen metabolism, in order to ensure optimal disposition of scarce resources. This is the first investigation of central metabolism for an oligotrophic ultramicrobacterium that possesses a relatively large genome size. In contrast to the behavior so far observed for SAR11 oligotrophic bacteria, S. alaskensis shows a physiological capacity to exploit increases in ambient nutrient availability and thereby achieve high-population densities. The ISME Journal (2009) 3, 1036-1052; doi:10.1038/ismej.2009.52; published online 21 May 2009 C1 [Williams, Timothy J.; Ertan, Haluk; Ting, Lily; Cavicchioli, Ricardo] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia. [Ertan, Haluk] Istanbul Univ, Dept Mol Biol & Genet, Fac Sci, Istanbul, Turkey. RP Cavicchioli, R (corresponding author), Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia. 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Mitchell, James G. Hu, Xiaoke Tang, Haitian Zhang, Hua Sheng, Yanqing TI Bacterial community structure in the Bohai Strait provides insights into organic matter niche partitioning SO CONTINENTAL SHELF RESEARCH DT Article AB The Bohai Strait is a crucial pathway for water exchange between the Bohai Sea and the Yellow Sea, and exhibits a robust spatiotemporal pattern of hydrodynamic conditions. 16S rRNA amplicon sequencing was applied to study the spatiotemporal patterns of bacterial community composition and diversity in the Bohai Strait. Physical and chemical parameters were measured in order to explain what might control the observed patterns in community composition and diversity. In response to environmental changes between seasons, especially the dramatic changes of temperature and nutrient (NO2-N and PO4-P) levels, Synechococcus and unclassified genera in Family I predominated in summer while some oligotrophic taxa (e.g., Pelagibacter, OM43 Glade) are more abundant in winter. The spatial heterogeneity and overall patchiness of bacterial assemblages in the Bohai Strait could not be well explained by the measured factors. However, the taxa with the most spatially variable presence and absence, e.g. Vibrionaceae, SAR11 and Sva0996 marine group, possess differentiated niches for utilization of organic matter (OM) and display a close relationship with the distribution of OM sources in this area, which may indicate the significance of OM sources to bacterial community structure. Our results suggest that local factors rather than regional factors, such as dispersal limited by hydrodynamics, structure the bacterial communities in the Bohai Strait. C1 [Wang, Yibo; Wang, Bin; Hu, Xiaoke] Chinese Acad Sci, Key Lab Coastal Biol & Bioresource Utilizat, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China. [Wang, Yibo; Wang, Bin; Hu, Xiaoke] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266237, Peoples R China. 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Amaral-Zettler, Linda A. Ducklow, Hugh W. Rich, Jeremy J. TI Seasonal Succession of Free-Living Bacterial Communities in Coastal Waters of the Western Antarctic Peninsula SO FRONTIERS IN MICROBIOLOGY DT Article AB The marine ecosystem along the Western Antarctic Peninsula undergoes a dramatic seasonal transition every spring, from almost total darkness to almost continuous sunlight, resulting in a cascade of environmental changes, including phytoplankton blooms that support a highly productive food web. Despite having important implications for the movement of energy and materials through this ecosystem, little is known about how these changes impact bacterial succession in this region. Using 16S rRNA gene amplicon sequencing, we measured changes in free-living bacterial community composition and richness during a 9-month period that spanned winter to the end of summer. Chlorophyll a concentrations were relatively low until summer when a major phytoplankton bloom occurred, followed 3 weeks later by a high peak in bacterial production. Richness in bacterial communities varied between similar to 1,200 and 1,800 observed operational taxonomic units (OTUs) before the major phytoplankton bloom (out of similar to 43,000 sequences per sample). During peak bacterial production, OTU richness decreased to similar to 700 OTUs. The significant decrease in OTU richness only lasted a few weeks, after which time OTU richness increased again as bacterial production declined toward pre-bloom levels. OTU richness was negatively correlated with bacterial production and chlorophyll a concentrations. Unlike the temporal pattern in OTU richness, community composition changed from winter to spring, prior to onset of the summer phytoplankton bloom. Community composition continued to change during the phytoplankton bloom, with increased relative abundance of several taxa associated with phytoplankton blooms, particularly Polaribacter. Bacterial community composition began to revert toward pre-bloom conditions as bacterial production declined. Overall, our findings clearly demonstrate the temporal relationship between phytoplankton blooms and seasonal succession in bacterial growth and community composition. Our study highlights the importance of high-resolution time series sampling, especially during the relatively under-sampled Antarctic winter and spring, which enabled us to discover seasonal changes in bacterial community composition that preceded the summertime phytoplankton bloom. C1 [Luria, Catherine M.] Brown Univ, Dept Ecol & Evolutionary Biol, Providence, RI 02912 USA. [Amaral-Zettler, Linda A.] Marine Biol Lab, Josephine Bay Paul Ctr Comparat Mol Biol & Evolut, Woods Hole, MA 02543 USA. 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TI Marine bacterial, archaeal and protistan association networks reveal ecological linkages SO ISME JOURNAL DT Article AB Microbes have central roles in ocean food webs and global biogeochemical processes, yet specific ecological relationships among these taxa are largely unknown. This is in part due to the dilute, microscopic nature of the planktonic microbial community, which prevents direct observation of their interactions. Here, we use a holistic (that is, microbial system-wide) approach to investigate time-dependent variations among taxa from all three domains of life in a marine microbial community. We investigated the community composition of bacteria, archaea and protists through cultivation-independent methods, along with total bacterial and viral abundance, and physicochemical observations. Samples and observations were collected monthly over 3 years at a well-described ocean time-series site of southern California. To find associations among these organisms, we calculated time-dependent rank correlations (that is, local similarity correlations) among relative abundances of bacteria, archaea, protists, total abundance of bacteria and viruses and physico-chemical parameters. We used a network generated from these statistical correlations to visualize and identify time-dependent associations among ecologically important taxa, for example, the SAR11 cluster, stramenopiles, alveolates, cyanobacteria and ammonia-oxidizing archaea. Negative correlations, perhaps suggesting competition or predation, were also common. The analysis revealed a progression of microbial communities through time, and also a group of unknown eukaryotes that were highly correlated with dinoflagellates, indicating possible symbioses or parasitism. Possible 'keystone' species were evident. The network has statistical features similar to previously described ecological networks, and in network parlance has non-random, small world properties (that is, highly interconnected nodes). This approach provides new insights into the natural history of microbes. The ISME Journal (2011) 5, 1414-1425; doi:10.1038/ismej.2011.24; published online 24 March 2011 C1 Univ So Calif, Dept Biol Sci, Los Angeles, CA 90089 USA. Univ So Calif, Wrigley Inst Environm Studies, Los Angeles, CA USA. RP Steele, JA (corresponding author), CALTECH, Div Geol & Planetary Sci, Mail Code 100-23, Pasadena, CA 91125 USA. 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It indicated that phage infection to the SAR11 cluster may occur ubiquitously and has significant impacts on bathypelagic SAR11 communities in the deep sea. Meanwhile, as Prochlorococcus is prominently distributed in the euphotic ocean, the existence of their potential phages in the deep sea suggested the sedimentation mechanism might contribute to the formation of the deep sea viromes. Intriguingly, the presence of Mycobacterium phages only in the deep sea viromes, suggests inhabitance of endemic viral populations in the deep sea viromes in the SCSV. This study provided an insight of the viral community in the South China Sea and for the first time uncovered the deep sea viral diversity in the central basin of the South China Sea. C1 [Liang, Yantao; Wang, Long; Wang, Zengmeng; Zhao, Jiulong; Zhang, Yongyu] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Shandong Prov Key Lab Energy Genet, Qingdao, Shandong, Peoples R China. 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Marine viral communities are composed of numerous genetically distinct subfamily/genus-level viral groups. Among currently identified viral groups, the HMO-2011-type group is known to be dominant and broadly distributed. However, only four HMO-2011-type cultivated representatives that infect marine SAR116 and Roseobacter strains have been reported to date, and the genetic diversity, potential hosts, and ecology of this group remain poorly elucidated. Here, we present the genomes of seven HMO-2011-type phages that were isolated using four Roseobacter strains and one SAR11 strain, as well as additional 207 HMO-2011-type metagenomic viral genomes (MVGs) identified from various marine viromes. Phylogenomic and shared-gene analyses revealed that the HMO-2011-type group is a subfamily-level group comprising at least 10 discernible genus-level subgroups. Moreover, >2000 HMO-2011-type DNA polymerase sequences were identified, and the DNA polymerase phylogeny also revealed that the HMO-2011-type group contains diverse subgroups and is globally distributed. Metagenomic read-mapping results further showed that most HMO-2011-type phages are prevalent in global oceans and display distinct geographic distributions, with the distribution of most HMO-2011-type phages being associated with temperature. Lastly, we found that members in subgroup IX, represented by pelagiphage HTVC033P, were among the most abundant HMO-2011-type phages, which implies that SAR11 bacteria are crucial hosts for this viral group. In summary, our findings substantially expand current knowledge regarding the phylogenetic diversity, evolution, and distribution of HMO-2011-type phages, highlighting HMO-2011-type phages as major ecological agents that can infect certain key bacterial groups. 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The cascade of environmental changes brought about by upwelling and relaxation events thus impacts microbial community structure in the bay, with important implications for the temporal variability of nutrient and energy fluxes within the MB ecosystem. Our observations emphasize the need for continued monitoring of planktonic microbial communities in order to predict and manage the behavior of this sensitive marine sanctuary ecosystem, over projected intensification of upwelling in the region. C1 [Reji, Linta; Tolar, Bradley B.; Francis, Christopher A.] Stanford Univ, Dept Earth Syst Sci, Stanford, CA 94305 USA. [Chavez, Francisco P.] Monterey Bay Aquarium Res Inst, Biol Oceanog Grp, Moss Landing, CA USA. RP Francis, CA (corresponding author), Stanford Univ, Dept Earth Syst Sci, Stanford, CA 94305 USA. 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We here review how observational, experimental, and theoretical evidence converge on the conclusion that the marine pelagic community of heterotrophic prokaryotes consists of organisms with two main types of life strategies. We illustrate this dichotomy by SAR11 and Vibrio spp. as typical representatives of the two strategies. A theory for life strategy dichotomy exists in classical r/K-selection. We here discuss an additional dichotomy introduced by what we term S/L-selection (for Small and Large, respectively). While r/K-selection focuses on the role of environmental disturbances, steady-state models suggest that high abundance at species level should be closely related to a low trade-off between competition and defense. We summarize literature indicating that the high availability of organic C is an essential environmental factor favoring Vibrio spp. and suggest that the essence of the generalized L-strategy is to reduce the competition-predator defense trade-off by using non-limiting organic C to increase size. The "streamlining" theory that has been suggested for the S-strategist SAR11 proposes the opposite: that low trade-off is achieved by a reduction in size. We show how this apparent contradiction disappears when the basic assumptions of diffusion-limited uptake are considered. We propose a classification scheme that combines S/L and r/K-selection using the two dimensions of organic C availability and environmental disturbance. As organic C in terrestrial runoff and size of the oligotrophic oceanic gyres are both changing, habitat size for both S- and L-strategists are affected by global change. 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Both the full mix of substances leached from polyvinyl chloride (PVC) and zinc alone had profound impacts on the taxonomic and functional diversity of our natural planktonic community. Microbial primary producers, both prokaryotic and eukaryotic, which comprise the base of the marine food web, were strongly impaired by exposure to plastic leachates, showing significant declines in photosynthetic efficiency, diversity, and abundance. Key heterotrophic taxa, such as SAR11, which are the most abundant planktonic organisms in the ocean, also exhibited significant declines in relative abundance when exposed to higher levels of PVC leachate. In contrast, many copiotrophic bacteria, including members of the Alteromonadales, dramatically increased in relative abundance under both exposure treatments. 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Ulloa, Osvaldo TI Oxygen modulates bacterial community composition in the coastal upwelling waters off central Chile SO DEEP-SEA RESEARCH PART II-TOPICAL STUDIES IN OCEANOGRAPHY DT Article; Proceedings Paper CT International Symposium on Microbial Responses to Ocean Deoxygenation CY DEC, 2016 CL Natl Inst Oceanog, Council Sci & Ind Res, INDIA HO Natl Inst Oceanog, Council Sci & Ind Res AB Bacterial community composition and its relationship to oxygen were investigated in the non-sulfidic shelf waters exposed to seasonal upwelling and oxygen deficiency off central Chile. Using 16S rRNA gene clone libraries, terminal restriction fragment length polymorphism (T-RFLP) and iTAG sequencing analyses of nearly 4 years of monthly sampling through the water column (5-80 m depth), we found a clear partitioning of community composition that could be attributed to dissolved oxygen (Spearman correlation coefficient = 0.578) in comparison to other environmental variables, such as light, fluorescence, temperature, salinity, microbial abundance or nutrients. Bacteroidetes (orders Sphingobacteriales and Flavobacteriales), SAR11 (subclades Ia - Candidatus Pelagibacter ubique and II), an unclassified group of Alphaproteobacteria, and SAR86 dominated in waters containing dissolved oxygen concentrations >= 70 mu M. In contrast, taxonomic groups associated with capabilities of either oxidative or reductive cycling of inorganic sulfur dominated in waters with dissolved oxygen from similar to 70 mu M to undetectable levels. The dominant groups were Arctic96BD-19, SUP05 and SAR324 (sulfur oxidation) and Desulfobacterales (sulfate reduction). Desulfobacterales are known to come primarily from sulfidic, nitrate/nitrite-depleted waters and sediments. Their presence, therefore, provides taxonomic evidence for pelagic dissimilatory sulfate reduction within oxygen-depleted coastal environments. C1 [Aldunate, Montserrat; Bertagnolli, Anthony D.; Ulloa, Osvaldo] Univ Concepcion, Dept Oceanog, Casilla 160-C, Concepcion 4070386, Chile. [Aldunate, Montserrat; Bertagnolli, Anthony D.; Ulloa, Osvaldo] Univ Concepcion, Inst Milenio Oceanog, Casilla 160-C, Concepcion 4070386, Chile. [Aldunate, Montserrat] Univ Concepcion, Programas Postgrado Oceanog, Casilla 160-C, Concepcion 4070386, Chile. [De la Iglesia, Rodrigo] Pontificia Univ Catolica Chile, Dept Mol Genet & Microbiol, Ave Libertador Gen Bernardo OHiggins 340, Santiago 8331150, Chile. 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Data demonstrated that this large viral microdiversity somehow mirrors the host species diversity since approximate to 50% of the 926 analyzed Pelagibacter genomes were found to belong to independent BSC species that do not significantly engage in gene flow with one another. The host range of this evolutionarily successful virus revealed that a single viral species can infect multiple Pelagibacter BSC species, indicating that this virus crosses not only formal BSC barriers but also biomes since viral ancestors are found in freshwater. C1 [Martinez-Hernandez, Francisco; Garcia-Heredia, Inmaculada; Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. [Diop, Awa; Bobay, Louis-Marie] Univ North Carolina Greensboro, Dept Biol, Greensboro, NC USA. RP Martinez-Garcia, M (corresponding author), Univ Alicante, Dept Physiol Genet & Microbiol, Alicante, Spain. 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[Jing, Xiaoyan; Gou, Honglei; Gong, Yanhai; Su, Xiaolu; Ji, Yuetong; Xu, Jian] Chinese Acad Sci, Qingdao Inst BioEnergy & Bioproc Technol, Shandong Key Lab Energy Genet, Qingdao, Shandong, Peoples R China. [Jing, Xiaoyan; Song, Yizhi; Thompson, Ian P.; Huang, Wei E.] Univ Oxford, Dept Engn Sci, Parks Rd, Oxford OX1 3PJ, England. [Jing, Xiaoyan; Gou, Honglei; Gong, Yanhai; Su, Xiaolu; Ji, Yuetong; Xu, Jian] Univ Chinese Acad Sci, Beijing, Peoples R China. [Xu, La] Marine Biol Inst Shandong Prov, Dis & Fishery Drugs Res Ctr, Qingdao, Shandong, Peoples R China. 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Alcolombri, Uria Uechi, Gen-Ichiro Jackson, Colin J. Laurino, Paola TI The ultra-high affinity transport proteins of ubiquitous marine bacteria SO NATURE DT Article AB SAR11 bacteria are the most abundant microorganisms in the surface ocean(1) and have global biogeochemical importance(2-4). To thrive in their competitive oligotrophic environment, these bacteria rely heavily on solute-binding proteins that facilitate uptake of specific substrates via membrane transporters(5,6). The functions and properties of these transport proteins are key factors in the assimilation of dissolved organic matter and biogeochemical cycling of nutrients in the ocean, but they have remained largely inaccessible to experimental investigation. Here we performed genome-wide experimental characterization of all solute-binding proteins in a prototypical SAR11 bacterium, revealing specific functions and general trends in their properties that contribute to the success of SAR11 bacteria in oligotrophic environments. We found that the solute-binding proteins of SAR11 bacteria have extremely high binding affinity (dissociation constant >20 pM) and high binding specificity, revealing molecular mechanisms of oligotrophic adaptation. Our functional data have uncovered new carbon sources for SAR11 bacteria and enable accurate biogeographical analysis of SAR11 substrate uptake capabilities throughout the ocean. This study provides a comprehensive view of the substrate uptake capabilities of ubiquitous marine bacteria, providing a necessary foundation for understanding their contribution to assimilation of dissolved organic matter in marine ecosystems. C1 [Clifton, Ben E.; Uechi, Gen-Ichiro; Laurino, Paola] Okinawa Inst Sci & Technol Grad Univ, Prot Engn & Evolut Unit, Onna, Japan. [Alcolombri, Uria] Hebrew Univ Jerusalem, Dept Plant & Environm Sci, Jerusalem, Israel. [Jackson, Colin J.] Australian Natl Univ, Res Sch Chem, Canberra, ACT, Australia. [Jackson, Colin J.] Australian Natl Univ, ARC Ctr Excellence Innovat Peptide & Prot Sci, Canberra, ACT, Australia. [Jackson, Colin J.] Australian Natl Univ, Res Sch Chem, ARC Ctr Excellence Synthet Biol, Canberra, ACT, Australia. [Laurino, Paola] Osaka Univ, Inst Prot Res, Suita, Osaka, Japan. RP Clifton, BE; Laurino, P (corresponding author), Okinawa Inst Sci & Technol Grad Univ, Prot Engn & Evolut Unit, Onna, Japan.; Laurino, P (corresponding author), Osaka Univ, Inst Prot Res, Suita, Osaka, Japan. 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Schwientek, Patrick Martinez-Garcia, Manuel Torrents, David McMahon, Katherine D. Andersson, Siv G. E. Stepanauskas, Ramunas Woyke, Tanja Bertilsson, Stefan TI Tuning fresh: radiation through rewiring of central metabolism in streamlined bacteria SO ISME JOURNAL DT Article AB Most free-living planktonic cells are streamlined and in spite of their limitations in functional flexibility, their vast populations have radiated into a wide range of aquatic habitats. Here we compared the metabolic potential of subgroups in the Alphaproteobacteria lineage SAR11 adapted to marine and freshwater habitats. Our results suggest that the successful leap from marine to freshwaters in SAR11 was accompanied by a loss of several carbon degradation pathways and a rewiring of the central metabolism. Examples for these are C1 and methylated compounds degradation pathways, the Entner-Doudouroff pathway, the glyoxylate shunt and anapleuretic carbon fixation being absent from the freshwater genomes. Evolutionary reconstructions further suggest that the metabolic modules making up these important freshwater metabolic traits were already present in the gene pool of ancestral marine SAR11 populations. The loss of the glyoxylate shunt had already occurred in the common ancestor of the freshwater subgroup and its closest marine relatives, suggesting that the adaptation to freshwater was a gradual process. Furthermore, our results indicate rapid evolution of TRAP transporters in the freshwater clade involved in the uptake of low molecular weight carboxylic acids. We propose that such gradual tuning of metabolic pathways and transporters toward locally available organic substrates is linked to the formation of subgroups within the SAR11 clade and that this process was critical for the freshwater clade to find and fix an adaptive phenotype. 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McLellan, Sandra L. Graham, Linda E. Young, Erica B. TI Invasive dreissenid mussels and benthic algae in Lake Michigan: characterizing effects on sediment bacterial communities SO FEMS MICROBIOLOGY ECOLOGY DT Article AB Dreissenid mussels have invaded the Laurentian Great Lakes causing dramatic changes to benthic-pelagic interactions. Despite research on food web impacts, there is limited data on mussel effects on benthic bacterial communities. This study examined effects of dreissenid mussels and benthic algae on sediment bacterial community composition and diversity. Triplicate experimental sediment plus lake water microcosms were used and either mussels, benthic algae or both were added. Changes in water nutrient chemistry and sediment bacterial communities were monitored using 16S rRNA amplicon sequencing, over 21 days. When mussels were present, nitrate and soluble reactive P increased significantly as the dominant N and P forms. Bacterial diversity increased in all microcosms, although bacterial community composition was distinct between treatment. Higher nitrate in mussel microcosms was accompanied by increases in nitrifying taxa (Nitrospira, Nitrosomonas), which are important in oxidizing mussel-excreted ammonium. Microcosms with algal additions showed increases in bacterial taxa capable of degrading algal cellulose, and Pelagibacter (SAR11) disappeared from all but control microcosms. This study suggests that bacterial communities in lake sediments respond to mussels and algae. Functional analysis of bacterial communities provides insights into changes in microbially mediated benthic nutrient transformations associated with invasive dreissenid mussels and benthic algae in lake ecosystems. C1 [Lee, Philip O.; Young, Erica B.] Univ Wisconsin, Dept Biol Sci, Milwaukee, WI 53201 USA. [McLellan, Sandra L.; Young, Erica B.] Univ Wisconsin, Sch Freshwater Sci, Milwaukee, WI 53204 USA. 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Pitre, David M. Weckhorst, Jessica Lee Lanclos, V. Celeste Webber, Austen T. Thrash, J. Cameron TI Artificial Seawater Media Facilitate Cultivating Members of the Microbial Majority from the Gulf of Mexico SO MSPHERE DT Article AB High-throughput cultivation studies have been successful at bringing numerous important marine bacterioplankton lineages into culture, yet these frequently utilize natural seawater media that can hamper portability, reproducibility, and downstream characterization efforts. Here we report the results of seven experiments with a set of newly developed artificial seawater media and evaluation of cultivation success via comparison with community sequencing data from the inocula. Eighty-two new isolates represent highly important marine clades, including SAR116, OM60/NOR5, SAR92, Roseobacter, and SAR11. For many, isolation with an artificial seawater medium is unprecedented, and several organisms are also the first of their type from the Gulf of Mexico. Community analysis revealed that many isolates were among the 20 most abundant organisms in their source inoculum. This method will expand the accessibility of bacterioplankton cultivation experiments and improve repeatability by avoiding normal compositional changes in natural seawater. IMPORTANCE The difficulty in cultivating many microbial taxa vexes researchers intent on understanding the contributions of these organisms to natural systems, particularly when these organisms are numerically abundant, and many cultivation attempts recover only rare taxa. Efforts to improve this conundrum with marine bacterioplankton have been successful with natural seawater media, but that approach suffers from a number of drawbacks and there have been no comparable artificial alternatives created in the laboratory. This work demonstrates that a newly developed suite of artificial seawater media can successfully cultivate many of the most abundant taxa from seawater samples and many taxa previously only cultivated with natural seawater media. This methodology therefore significantly simplifies efforts to cultivate bacterioplankton and greatly improves our ability to perform physiological characterization of cultures postisolation. C1 [Henson, Michael W.; Pitre, David M.; Weckhorst, Jessica Lee; Lanclos, V. Celeste; Webber, Austen T.; Thrash, J. Cameron] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA. RP Thrash, JC (corresponding author), Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA. 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Art-science collaborations play an integral role in this. Traditionally, these involve pairing artists and marine scientists to work on a project which is then presented to an audience. Increasingly however more interactive relationships with the public are being seen as a beneficial way to merge art and scientific data. The Ocean Science Jam is such a project that brings musicians, artists, dancers, performers, and the public together to respond creatively in real time to visual and audio cues based on a theme related to marine scientists' work. By mixing creativity with science in an integrative way the Ocean Science Jam not only acts as tool for public communication but also opens new ways for scientific data to be interpreted by non-scientists. This paper will explore this initiative from design to delivery highlighting the results of facilitating with the public moments where new responses to ocean science can be created through art. 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Widdicombe, Claire E. Tarran, Glen A. Polimene, Luca Dashfield, Sarah Beale, Rachael Scanlan, David J. Chen, Yin TI Microbial uptake dynamics of choline and glycine betaine in coastal seawater SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Choline and glycine betaine (GBT) are utilized as osmolytes to counteract osmotic stress, but also constitute important nutrient sources for many marine microbes. Bacterial catabolism of these substrates can then lead to the production of climate active trace gases such as methylamine and methane. Using radiotracers, we investigated prokaryotic choline/GBT uptake and determined biotic and abiotic factors driving these processes in the Western English Channel, UK. Kinetic uptake parameters indicated high affinity (nM range) for both osmolytes and showed a seasonal pattern for choline uptake. Generalized linear modeling of uptake parameters suggested a significant influence of sea surface temperature and salinity on prokaryotic uptake of both osmolytes. The presence of diatoms significantly influenced prokaryotic choline/GBT uptake dynamics. Choline uptake was further related to the occurrence of Phaeocystis spp., which were highly abundant in the phytoplankton community during spring, and dinoflagellates abundance during summer. While Rhodobacteraceae were the most important bacterial drivers for prokaryotic choline uptake, prokaryotic GBT uptake was associated with various groups such as SAR11 (Pelagibacterales) and Gammaproteobacteria, suggesting a wider capacity for GBT catabolism than previously recognized. Furthermore, using a newly developed approach we determined the first available data for dissolved GBT concentrations in seawater and found both osmolytes to be at the sub-nanomolar range. Together, this study improves our understanding of the biogeochemical cycling of these environmentally important osmolytes and highlights how their cycles may be affected by a changing climate. 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Jones, Leandro R. TI Are ocean currents to slow to counteract SAR11 evolution? A next-generation sequencing, phylogeographic analysis (vol 107, pg 324, 2017) SO MOLECULAR PHYLOGENETICS AND EVOLUTION DT Correction C1 Consejo Nacl Invest Cient & Tecn, Av Rivadavia 1917 C1083ACA, Buenos Aires, DF, Argentina. [Jones, Leandro R.] Univ Nacl Patagonia San Juan Bosco, Fac Ciencias Nat Sede Trelew, Lab Virol & Genet Mol, 9 Julio & Belgrano S-N 9100, Trelew, Chubut, Argentina. RP Jones, LR (corresponding author), Univ Nacl Patagonia San Juan Bosco, Fac Ciencias Nat Sede Trelew, Lab Virol & Genet Mol, 9 Julio & Belgrano S-N 9100, Trelew, Chubut, Argentina. EM ljones@conicet.gov.ar CR Manrique JM, 2017, MOL PHYLOGENET EVOL, V107, P324, DOI 10.1016/j.ympev.2016.11.015 NR 1 TC 0 Z9 0 PD MAY PY 2017 VL 110 BP 150 EP 150 DI 10.1016/j.ympev.2017.03.001 UT WOS:000398877500015 DA 2025-07-30 ER PT J AU Mizuno, CM Ghai, R Saghaï, A López-García, P Rodriguez-Valera, F AF Mizuno, Carolina Megumi Ghai, Rohit Saghai, Aurelien Lopez-Garcia, Purificacion Rodriguez-Valera, Francisco TI Genomes of Abundant and Widespread Viruses from the Deep Ocean SO MBIO DT Article AB The deep sea is a massive, largely oligotrophic ecosystem, stretched over nearly 65% of the planet's surface. Deep-sea planktonic communities are almost completely dependent upon organic carbon sinking from the productive surface, forming a vital component of global biogeochemical cycles. However, despite their importance, viruses from the deep ocean remain largely unknown. Here, we describe the first complete genomes of deep-sea viruses assembled from metagenomic fosmid libraries. "Candidatus Pelagibacter" (SAR11) phage HTVC010P and Puniceispirillum phage HMO-2011 are considered the most abundant cultured marine viruses known to date. Remarkably, some of the viruses described here recruited as many reads from deep waters as these viruses do in the photic zone, and, considering the gigantic scale of the bathypelagic habitat, these genomes provide information about what could be some of the most abundant viruses in the world at large. Their role in the viral shunt in the global ocean could be very significant. Despite the challenges encountered in inferring the identity of their hosts, we identified one virus predicted to infect members of the globally distributed SAR11 cluster. We also identified a number of putative proviruses from diverse taxa, including deltaproteobacteria, bacteroidetes, SAR11, and gammaproteobacteria. Moreover, our findings also indicate that lysogeny is the preferred mode of existence for deep-sea viruses inhabiting an energy-limited environment, in sharp contrast to the predominantly lytic lifestyle of their photic-zone counterparts. Some of the viruses show a widespread distribution, supporting the tenet "everything is everywhere" for the deep-ocean virome. IMPORTANCE The deep sea is among the largest known habitats and a critical cog in biogeochemical cycling but remains under-explored in its microbiology. Even more than is the case for its prokaryotic community, our knowledge of its viral component has remained limited by the paucity of information provided by studies dependent upon short sequence fragments. In this work, we attempt to fill this existing gap by using a combination of classical fosmid libraries with next-generation sequencing and assembly to recover long viral genomic fragments. We have sequenced ca. 6,000 fosmids from two metagenomics libraries made from prokaryotic biomass from the deep Mediterranean Sea and recovered twenty-eight complete viral genomes, all of them novel and quite distinct from all previously described viral genomes. They are preferentially found in deeper waters and are widely distributed all over the oceans. To our knowledge, this is the first report on complete and cosmopolitan viral genomes from the bathypelagic habitat. C1 [Mizuno, Carolina Megumi; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Evolutionary Genom Grp, Alicante, Spain. [Mizuno, Carolina Megumi] Inst Pasteur, Dept Microbiol, Unit Mol Biol Gene Extremophiles, Paris, France. [Ghai, Rohit] Acad Sci Czech Republ, Inst Hydrobiol, Dept Aquat Microbial Ecol, Ctr Biol, Ceske Budejovice, Czech Republic. [Saghai, Aurelien; Lopez-Garcia, Purificacion] Univ Paris 11, CNRS UMR 8079, Unite Ecol Systemat & Evolut, Orsay, France. RP Rodriguez-Valera, F (corresponding author), Univ Miguel Hernandez, Evolutionary Genom Grp, Alicante, Spain. 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However, studies of viral communities are challenging to interpret because the vast majority of viruses are known only from sequence fragments, and their taxonomy, hosts, and genetic repertoires are unknown. Milne Fiord, located on the coastal margin of the Last Ice Area (LIA) in the High Arctic (82 degrees N, Canada), harbors an epishelf lake, a rare type of ice-dependent ecosystem in which a layer of freshwater overlies marine water connected to the open ocean. This microbe-dominated ecosystem faces catastrophic change due to the deterioration of its ice environment related to warming temperatures. We produced the first assessment of viral abundance, diversity, and distribution in this vulnerable ecosystem and explored the niches available for viral taxa and the functional genes underlying their distribution. We found that the viral community in the freshwater layer was distinct from, and more diverse than, the community in the underlying seawater and contained a different set of putative auxiliary metabolic genes, including the sulfur starvation-linked gene tauD and the gene coding for patatin-like phospholipase. The halocline community resembled the freshwater more than the marine community, but harbored viral taxa unique to this layer. We observed distinct viral assemblages immediately below the halocline, at a depth that was associated with a peak of prasinophyte algae and the viral family Phycodnaviridae. We also assembled 15 complete circular genomes, including a putative Pelagibacter phage with a marine distribution. It appears that despite its isolated and precarious situation, the varied niches in this epishelf lake support a diverse viral community, highlighting the importance of characterizing underexplored microbiota in the Last Ice Area before these ecosystems undergo irreversible change. IMPORTANCE Viruses are key to understanding polar aquatic ecosystems, which are dominated by microorganisms. However, studies of viral communities are challenging to interpret because the vast majority of viruses are known only from sequence fragments, and their taxonomy, hosts, and genetic repertoires are unknown. Our study establishes a basis for comparison that will advance understanding of viral ecology in diverse global environments, particularly in the High Arctic. Rising temperatures in this region mean that researchers have limited time remaining to understand the biodiversity and biogeochemical cycles of ice-dependent environments and the consequences of these rapid, irreversible changes. 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C1 [Saini, Priya; Wani, Shadil Ibrahim; Kumar, Ranjai; Chhabra, Ravneet; Sareen, Dipti] Panjab Univ, Dept Biochem, Chandigarh 160014, India. [Chimni, Swapandeep Singh] Guru Nanak Dev Univ, Dept Chem, Amritsar 143005, Punjab, India. RP Sareen, D (corresponding author), Panjab Univ, Dept Biochem, Sect 14, Chandigarh 160014, India. EM diptsare@pu.ac.in CR Saini P, 2014, PROTEIN EXPRES PURIF, V104, P71, DOI 10.1016/j.pep.2014.09.004 NR 1 TC 0 Z9 0 PD AUG PY 2015 VL 112 BP 50 EP 50 DI 10.1016/j.pep.2014.10.011 UT WOS:000355572200008 DA 2025-07-30 ER PT J AU Teira, E Martínez-García, S Lonborg, C Alvarez-Salgado, XA AF Teira, Eva Martinez-Garcia, Sandra Lonborg, Christian Alvarez-Salgado, Xose A. TI Growth rates of different phylogenetic bacterioplankton groups in a coastal upwelling system SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB Microbial degradation of dissolved organic matter (DOM) in planktonic ecosystems is carried out by diverse prokaryotic communities, whose growth rates and patterns of DOM utilization modulate carbon and nutrient biogeochemical cycles at local and global scales. Nine dilution experiments (September 2007 to June 2008) were conducted with surface water from the highly productive coastal upwelling system of the R a de Vigo (NW Iberian Peninsula) to estimate bacterial growth rates of six relevant marine bacterial groups: Roseobacter, SAR11, Betaproteobacteria, Gammaproteobacteria, SAR86 and Bacteroidetes. Surprisingly, SAR11 dominated over the other bacterial groups in autumn, likely associated to the entry of nutrient-rich, DOC-poor Eastern North Atlantic Central Water (ENACW) into the embayment. Roseobacter and SAR11 showed significantly opposing growth characteristics. SAR11 consistently grows at low rates (range 0.19-0.71 day(-1)), while Roseobacter has a high growth potential (range 0.70-1.64 day(-1)). In contrast, Betaproteobacteria, Bacteroidetes, SAR86 and Gammaproteobacteria growth rates widely varied among experiments. 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Here we show that three new enzymes related to fatty acid beta-oxidation constitute the pathway that assimilates methylmercaptopropionate (MMPA), the first product of DMSP demethylation/demethiolation, and that two previously unknown coenzyme A (CoA) derivatives, 3-methylmercaptopropionyl-CoA (MMPA-CoA) and methylthioacryloyl-CoA (MTA-CoA), are formed as novel intermediates. A member of the marine roseobacters, Ruegeria pomeroyi DSS-3, requires the MMPA-CoA pathway for MMPA assimilation and MeSH production. This pathway and the ability to produce MeSH from MMPA are present in diverse bacteria, and the ubiquitous SAR11 clade bacterium Pelagibacter ubique possesses enzymes for at least the first two steps. Analysis of marine metagenomic data indicates that the pathway is widespread among bacterioplankton in the ocean surface waters, making it one of the most important known routes for acquisition of reduced carbon and sulphur by surface ocean heterotrophs. 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The two forms of DmdB present in the marine roseobacter Ruegeria pomeroyi DSS-3, RPO_DmdB1 and RPO_DmdB2, and the single form in the SAR11 clade bacterium "Candidatus Pelagibacter ubique" HTCC1062, PU_DmdB1, were characterized in detail. DmdB enzymes were also examined from Ruegeria lacuscaerulensis ITI-1157, Pseudomonas aeruginosa PAO1, and Burkholderia thailandensis E264. The DmdB enzymes separated into two phylogenetic clades. All enzymes had activity with MMPA and were sensitive to inhibition by salts, but there was no correlation between the clades and substrate specificity or salt sensitivity. All Ruegeria species enzymes were inhibited by physiological concentrations (70 mM) of DMSP. However, ADP reversed the inhibition of RPO_DmdB1, suggesting that this enzyme was responsive to cellular energy charge. MMPA reversed the inhibition of RPO_DmdB2 as well as both R. lacuscaerulensis ITI-1157 DmdB enzymes, suggesting that a complex regulatory system exists in marine bacteria. 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Millar, Justin J. Payne, Jason T. Ochs, Clifford A. TI Free-Living and Particle-Associated Bacterioplankton in Large Rivers of the Mississippi River Basin Demonstrate Biogeographic Patterns SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB The different drainage basins of large rivers such as the Mississippi River represent interesting systems in which to study patterns in freshwater microbial biogeography. Spatial variability in bacterioplankton communities in six major rivers (the Upper Mississippi, Missouri, Illinois, Ohio, Tennessee, and Arkansas) of the Mississippi River Basin was characterized using Ion Torrent 16S rRNA amplicon sequencing. 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Muratore, Daniel Newton, Ryan J. Coleman, Maureen L. TI Reevaluating the Salty Divide: Phylogenetic Specificity of Transitions between Marine and Freshwater Systems SO MSYSTEMS DT Article AB Marine and freshwater microbial communities are phylogenetically distinct, and transitions between habitat types are thought to be infrequent. We compared the phylogenetic diversity of marine and freshwater microorganisms and identified specific lineages exhibiting notably high or low similarity between marine and freshwater ecosystems using a meta-analysis of 16S rRNA gene tag-sequencing data sets. As expected, marine and freshwater microbial communities differed in the relative abundance of major phyla and contained habitat-specific lineages. At the same time, and contrary to expectations, many shared taxa were observed in both habitats. Based on several metrics, we found that Gammaproteobacteria, Alphaproteobacteria, Bacteroidetes, and Betaproteobacteria contained the highest number of closely related marine and freshwater sequences, suggesting comparatively recent habitat transitions in these groups. Using the abundant alphaproteobacterial group SAR11 as an example, we found evidence that new lineages, beyond the recognized LD12 Glade, are detected in freshwater at low but reproducible abundances; this evidence extends beyond the 16S rRNA locus to core genes throughout the genome. Our results suggest that shared taxa are numerous, but tend to occur sporadically and at low relative abundance in one habitat type, leading to an underestimation of transition frequency between marine and freshwater habitats. Rare taxa with abundances near or below detection, including lineages that appear to have crossed the salty divide relatively recently, may possess adaptations enabling them to exploit opportunities for niche expansion when environments are disturbed or conditions change. IMPORTANCE The distribution of microbial diversity across environments yields insight into processes that create and maintain this diversity as well as potential to infer how communities will respond to future environmental changes. We integrated data sets from dozens of freshwater lake and marine samples to compare diversity across open water habitats differing in salinity. Our novel combination of sequence-based approaches revealed lineages that likely experienced a recent transition across habitat types. These taxa are promising targets for studying physiological constraints on salinity tolerance. 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TI Testing the metabolic theory of ecology with marine bacteria: different temperature sensitivity of major phylogenetic groups during the spring phytoplankton bloom SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Although temperature is a key driver of bacterioplankton metabolism, the effect of ocean warming on different bacterial phylogenetic groups remains unclear. Here, we conducted monthly short-term incubations with natural coastal bacterial communities over an annual cycle to test the effect of experimental temperature on the growth rates and carrying capacities of four phylogenetic groups: SAR11, Rhodobacteraceae, Gammaproteobacteria and Bacteroidetes. SAR11 was the most abundant group year-round as analysed by CARD-FISH, with maximum abundances in summer, while the other taxa peaked in spring. All groups, including SAR11, showed high temperature-sensitivity of growth rates and/or carrying capacities in spring, under phytoplankton bloom or post-bloom conditions. 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We revisited previous investigations in this area using concatenated alignments of the small and large subunit (SSU and LSU) rRNA genes, as we show here that these loci have much lower GC bias than whole genomes. This approach has allowed us to update the canonical 16S rRNA gene tree of the Alphaproteobacteria with additional important taxa that were not previously included, and with added resolution provided by concatenating the SSU and LSU genes. We investigated the topological stability of the Alphaproteobacteria by varying alignment methods, rate models, taxon selection and RY-recoding to circumvent GC content bias. We also introduce RYMK-recoding and show that it avoids some of the information loss in RY-recoding. We demonstrate that the topology of the Alphaproteobacteria is sensitive to inclusion of several groups of taxa, but it is less affected by the choice of alignment and rate methods. 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The base composition of their DNA is often highly biased towards low G+C content, a possible source of systematic error in phylogenetic reconstruction. A total of 228 orthologous protein families were sampled that are shared among major lineages of Alphaproteobacteria, including the marine free-living SAR11 clade and the obligate endosymbiotic Rickettsiales. These two ecologically distinct lineages share genome sizes of <1.5Mbp and genomic G+C content of <30%. Statistical analyses showed that only 28 protein families are composition-homogeneous, whereas the other 200 families significantly violate the composition-homogeneous assumption included in most phylogenetic methods. RAxML analysis based on the concatenation of 24 ribosomal proteins that fall into the heterogeneous protein category clustered the SAR11 and Rickettsiales lineages at the base of the Alphaproteobacteria tree, whereas that based on the concatenation of 28 homogeneous proteins (including 19 ribosomal proteins) disassociated the lineages and placed SAR11 at the base of the non-endosymbiotic lineages. When the two data sets were concatenated, only a model that accounted for compositional bias yielded a tree identical to the tree built with composition-homogeneous proteins. Ancestral genome analysis suggests that the first evolved SAR11 cell had a small genome streamlined from its ancestor by a factor of two and coinciding with an ecological transition, followed by further gradual streamlining towards the extant SAR11 populations. C1 [Luo, Haiwei] Chinese Univ Hong Kong, Sch Life Sci, Simon FS Li Marine Sci Lab, Shatin, Hong Kong, Peoples R China. 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Needham, David M. Fuhrman, Jed A. TI Every base matters: assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Microbial community analysis via high-throughput sequencing of amplified 16S rRNA genes is an essential microbiology tool. We found the popular primer pair 515F (515F-C) and 806R greatly underestimated (e.g. SAR11) or overestimated (e.g. Gammaproteobacteria) common marine taxa. We evaluated marine samples and mock communities (containing 11 or 27 marine 16S clones), showing alternative primers 515F-Y (5'-GTGYCAGCMGCCGCGG TAA) and 926R (5'-CCGYCAATTYMTTTRAGTTT) yield more accurate estimates of mock community abundances, produce longer amplicons that can differentiate taxa unresolvable with 515F-C/806R, and amplify eukaryotic 18S rRNA. 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D'Hondt, Sofie Willems, Anne Airoldi, Laura De Clerck, Olivier TI Diversity and Temporal Dynamics of the Epiphytic Bacterial Communities Associated with the Canopy-Forming Seaweed Cystoseira cornpressa (Esper) Gerloff and Nizamuddin SO FRONTIERS IN MICROBIOLOGY DT Article AB Canopy-forming seaweed species of the genus Cystoseira form diverse and productive habitats along temperate rocky coasts of the Mediterranean Sea. Despite numerous studies on the rich macrofauna and flora associated with Cystoseira spp., there is little knowledge about the epiphytic bacteria. We analyzed bacterial populations associated with canopies of Cystoseira compressa, over an annual vegetative cycle (May-October), and their relationships with the bacterial populations in the surrounding seawater, at intertidal rocky shores in Vasto (Chieti-Italy). The bacterial diversity was assessed using Illumina Miseq sequences of V1-V3 hypervariable regions of 16S rRNA gene. C. compressa bacterial community was dominated by sequences of Proteobacteria and Bacteroidetes, Verrucomicrobia, Actinobacteria. and Cyanobactena especially of the Rhodobactenaceae, Flayobactenaceae, Sapropiraceae, Verrucomicrobiaceae, and Phyllobactenaceae families. Seawater libraries were also dominated by Proteobacteria and Bacteroidetes sequences, especially of the Candidatus Pelagibacter (SAR11) and Rhodobacteriaceae families, but were shown to be clearly distinct from C. compressa libraries with only few species in common between the two habitats. We observed a clear successional pattern in the epiphytic bacteria of C. compressa over time. These variations were characterized by gradual addition of OTUs (Verrucomicrobia, Actinobacteria and SR1) to the community over a growing season, indicative of a temporal gradient, rather than a radical reorganization of the bacterial community. Moreover, we also found an increase in abundance over time of Rhodobacteraceae, comprising six potential pathogenic genera, Ruegena, Nautella, Aquimarina, Loktanella, Saprospira, and Phaeobacter which seemed to be associated to aged thalli of C. compressa. These bacteria could have the potential to affect the health and ecology of the algae, suggesting the hypothesis of a possible, but still unexplored, role of the microbial communities in contributing to the extensive ongoing declines of populations of Cystoseira spp. in the Mediterranean Sea. C1 [Mancuso, Francesco P.; Airoldi, Laura] Univ Bologna, Ctr Interdipartimentale Ric Sci Ambientali, Dipartimento Sci Biol Geol & Ambientali, UO CoNISMa, Ravenna, Italy. [Mancuso, Francesco P.; D'Hondt, Sofie; De Clerck, Olivier] Univ Ghent, Phycol Res Grp, B-9000 Ghent, Belgium. [Mancuso, Francesco P.; D'Hondt, Sofie; De Clerck, Olivier] Univ Ghent, Ctr Mol Phylogenet & Evolut, B-9000 Ghent, Belgium. [Willems, Anne] Univ Ghent, Dept Biochem & Microbiol, Lab Microbiol, B-9000 Ghent, Belgium. RP Mancuso, FP; Airoldi, L (corresponding author), Univ Bologna, Ctr Interdipartimentale Ric Sci Ambientali, Dipartimento Sci Biol Geol & Ambientali, UO CoNISMa, Ravenna, Italy.; Mancuso, FP (corresponding author), Univ Ghent, Phycol Res Grp, B-9000 Ghent, Belgium.; Mancuso, FP (corresponding author), Univ Ghent, Ctr Mol Phylogenet & Evolut, B-9000 Ghent, Belgium. 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R. Lundin, Daniel Andersson, Anders F. Labrenz, Matthias Juergens, Klaus TI Phylogenetic Signals of Salinity and Season in Bacterial Community Composition Across the Salinity Gradient of the Baltic Sea SO FRONTIERS IN MICROBIOLOGY DT Article AB Understanding the key processes that control bacterial community composition has enabled predictions of bacterial distribution and function within ecosystems. In this study, we used the Baltic Sea as a model system to quantify the phylogenetic signal of salinity and season with respect to bacterioplankton community composition. The abundances of 16S rRNA gene amplicon sequencing reads were analyzed from samples obtained from similar geographic locations in July and February along a brackish to marine salinity gradient in the Baltic Sea. While there was no distinct pattern of bacterial richness at different salinities, the number of bacterial phylotypes in winter was significantly higher than in summer. 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However, the metabolic activities and contribution of actively growing prokaryotes to ecosystem energy fluxes remain underexplored. Here we describe the temporal and spatial dynamics of active prokaryotes in the different water masses of the Mediterranean Sea using a combination of bromodeoxyuridine labelling and 16S rRNA gene Illumina sequencing. Bulk and actively dividing prokaryotic communities were drastically different and depth stratified. Alteromonadales were rare in bulk communities (contributing 0.1% on average) but dominated the actively dividing community throughout the overall water column (28% on average). Moreover, temporal variability of actively dividing Alteromonadales oligotypes was evinced. SAR86, Actinomarinales and Rhodobacterales contributed on average 3-3.4% each to the bulk and 11, 8.4 and 8.5% to the actively dividing communities in the epipelagic zone, respectively. 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We herein present the design and preclinical evaluation of two novel NOTA/NODAGA-containing peptides suitable for labeling with the positron emission tomography (PET) radionuclide Ga-68. These analogs are based on the previously reported GRPR-antagonist DOTAGA-PEG2-[Sar(11)]RM26, developed for targeted radiotheraostic applications. Both NOTA-PEG2-[Sar(11)]RM26 and NODAGA-PEG2-[Sar(11)]RM26 were successfully labeled with Ga-68 and evaluated in vitro and in vivo using PC-3 cell models. Both, [Ga-68]Ga-NOTA-PEG2-[Sar(11)]RM26 and [Ga-68]Ga-NODAGA-PEG2-[Sar(11)]RM26 displayed high metal-chelate stability in phosphate buffered saline and against the EDTA-challenge. The two [Ga-68]Ga-labeled conjugates demonstrated highly GRPR-mediated uptake in vitro and in vivo and exhibited a slow internalization over time, typical for radioantagonistis. The [Ga-nat]Ga-loaded peptides displayed affinity in the low nanomole range for GRPR in competition binding experiments. The new radiotracers demonstrated biodistribution profiles suitable for diagnostic imaging shortly after administration with fast background clearance. Their high tumor uptake (13 +/- 1 and 15 +/- 3% IA/g for NOTA and NODAGA conjugates, respectively) and high tumor-to-blood ratios (60 +/- 10 and 220 +/- 70, respectively) 3 h pi renders them promising PET tracers for use in patients. Tumor-to-normal organ ratios were higher for [Ga-68]Ga-NODAGA-PEG2-[Sar(11)]RM26 than for the NOTA-containing counterpart. The performance of the two radiopeptides was further supported with the PET/CT images. In conclusion, [Ga-68]Ga-NODAGA-PEG2-[Sar(11)]RM26 is a promising PET imaging tracer for visualization of GRPR-expressing lesions with high imaging contrast shortly after administration. C1 [Kanellopoulos, Panagiotis; Bezverkhniaia, Ekaterina; Abouzayed, Ayman; Rosenstro''m, Ulrika; Orlova, Anna] Uppsala Univ, Dept Med Chem, S-75123 Uppsala, Sweden. 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Desantis, Todd Z. Saunders, F. Michael Andersen, Gary L. Liu, Wen-Tso TI Diversity of bacterioplankton in contrasting Tibetan lakes revealed by high-density microarray and clone library analysis SO FEMS MICROBIOLOGY ECOLOGY DT Article AB Tibetan lakes represent a unique microbial environment and are a good ecosystem to investigate the microbial diversity of high mountain lakes and their relationship with environmental factors. The diversity and community structure of bacterioplankton in Tibetan lakes was determined using DNA fingerprinting analysis, high-density 16S rRNA gene microarray (PhyloChip) analysis, and extensive clone library analysis of bacterial 16S rRNA genes. A previously unseen high microbial diversity (1732 operational taxonomic units based on PhyloChip data) and numerous novel bacterial 16S rRNA gene sequences were observed. Abundant SAR11-like sequences retrieved from saline Lake Qinghai demonstrated a unique SAR11 phylogenetic sister clade related to the freshwater LD12 clade. Water chemistry (e.g. salinity) and altitude played important roles in the selection of bacterial taxa (both presence and relative abundance) in Tibetan lakes. The ubiquity and uniqueness of bacterial taxa, as well as the correlation between environmental factors and bacterial taxa, was observed to vary gradually with different phylogenetic levels. Our study suggested high microbial cosmopolitanism and high endemicity observed at higher and lower phylogenetic levels, respectively. C1 [Zhang, Rui; Wu, Qinglong; Saunders, F. Michael; Liu, Wen-Tso] Natl Univ Singapore, Div Environm Sci & Engn, Singapore 117548, Singapore. [Zhang, Rui] Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen, Peoples R China. 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Thrash, J. Cameron Dupont, Chris L. Barbeau, Katherine A. TI Trace Metal Acquisition by Marine Heterotrophic Bacterioplankton with Contrasting Trophic Strategies SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Heterotrophic bacteria in the SAR11 and Roseobacter lineages shape the marine carbon, nitrogen, phosphorous, and sulfur cycles, yet they do so having adopted divergent ecological strategies. Currently, it is unknown whether these globally significant groups partition into specific niches with respect to micronutrients (e.g., trace metals) and how that may affect marine trace metal cycling. Here, we used comparative genomics to identify diverse iron, cobalt, nickel, copper, and zinc uptake capabilities in SAR11 and Roseobacter genomes and uncover surprising unevenness within and between lineages. The strongest predictors for the extent of the metal uptake gene content are the total number of transporters per genome, genome size, total metal transporters, and GC content, but numerous exceptions exist in both groups. Taken together, our results suggest that SAR11 have strongly minimized their trace metal uptake versatility, with high-affinity zinc uptake being a unique exception. The larger Roseobacter genomes have greater trace metal uptake versatility on average, but they also appear to have greater plasticity, resulting in phylogenetically similar genomes having largely different capabilities. Ultimately, phylogeny is predictive of the diversity and extent of 20 to 33% of all metal uptake systems, suggesting that specialization in metal utilization mostly occurred independently from overall lineage diversification in both SAR11 and Roseobacter. We interpret these results as reflecting relatively recent trace metal niche partitioning in both lineages, suggesting that concentrations and chemical forms of metals in the marine environment are important factors shaping the gene content of marine heterotrophic Alphaproteobacteria of the SAR11 and Roseobacter lineages. C1 [Hogle, Shane L.; Barbeau, Katherine A.] Univ Calif San Diego, Scripps Inst Oceanog, Geosci Res Div, La Jolla, CA 92093 USA. [Thrash, J. Cameron] Louisiana State Univ, Dept Biol Sci, Baton Rouge, LA 70803 USA. [Dupont, Chris L.] J Craig Venter Inst, La Jolla, CA USA. RP Hogle, SL (corresponding author), Univ Calif San Diego, Scripps Inst Oceanog, Geosci Res Div, La Jolla, CA 92093 USA. 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Sestanovic, Stefanija TI Contribution of different bacterial groups in the carbon flow through the microbial food web SO ESTUARINE COASTAL AND SHELF SCIENCE DT Article AB Bacteria greatly contributes to the transfer of dissolved organic carbon to the upper trophic levels. Nevertheless, little is known about the contribution of specific bacterial groups. Here, we conducted three seasonal experiments (both, microcosm and mesocosm) in the coastal area of the central Adriatic with the aim of determining the contribution of selected bacterial groups to carbon flow through the microbial food web. We assessed the growth rates of four bacterial groups (Bacteroidota, SAR11, Rhodobacteraceae, and Gammaproteobacteria), as well as aerobic anoxygenic phototrophs, and their contributions to biomass production and carbon transfer to upper trophic levels. For the first time, we reported a significant contribution of aerobic anoxygenic phototrophs to the carbon flow, especially during the summer. Under initial winter conditions, SAR11 contributed the most to bacterial biomass production, while Gammaproteobacteria were the primary contributors to grazing loss and served as the preferred prey. As temperature increased, the contribution of Gammaproteobacteria and Bacteroidota to bacterial biomass production also increased. Regarding grazing loss, SAR11 and Bacteroidota were the dominant contributors in spring, whereas Gammaproteobacteria and Rhodobacteraceae played the most significant role during summer. Under nutrientenriched conditions, SAR11 contributed the most to both bacterial biomass production and grazing loss during winter. However, with rising temperatures, Gammaproteobacteria, Rhodobacteraceae and Bacteroidetes became the primary contributors to bacterial biomass production and Gammaproteobacteria and Bacteroidetes in grazing loss. Our estimates suggest that Gammaproteobacteria and the SAR11 clade were more significantly impacted by grazing, whereas Bacteroidota and Rhodobacteraceae were equally influenced by both nutrient availability and grazing pressure. 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DeLong, Edward F. TI Metatranscriptomics reveals unique microbial small RNAs in the ocean's water column SO NATURE DT Article AB Microbial gene expression in the environment has recently been assessed via pyrosequencing of total RNA extracted directly from natural microbial assemblages. Several such 'metatranscriptomic' studies(1,2) have reported that many complementary DNA sequences shared no significant homology with known peptide sequences, and so might represent transcripts from uncharacterized proteins. Here we report that a large fraction of cDNA sequences detected in microbial metatranscriptomic data sets are comprised of well-known small RNAs ( sRNAs) 3, as well as new groups of previously unrecognized putative sRNAs ( psRNAs). These psRNAs mapped specifically to intergenic regions of microbial genomes recovered from similar habitats, displayed characteristic conserved secondary structures and were frequently flanked by genes that indicated potential regulatory functions. 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Particularly, bacterial contributions to phytoplankton-derived dimethylsulfoniopropionate (DMSP) degradation have been recently evaluated by using advanced sequencing technologies to understand their role in the marine microbial food web. Here, we surveyed the bacterial diversity and community composition under seasonal water mixing in the bay of Gwangyang (GW), a semi-enclosed estuary at the southern tip of the Korea Peninsula. We detected phylogenetic dissimilarities among season-specific habitats in GW and their specific bacterial taxa. Additionally, bacterial contribution to degradation of phytoplankton-derived DMSP from estuarine to coastal waters at euphotic depths in GW was investigated as the presence or absence of DMSP demethylation gene, encoded by dmdA. Among the operational taxonomic units (OTUs) in GW bacterial communities, the most dominant and ubiquitous OTU1 was affiliated with the SAR11 clade (SAR11-OTU). The population dynamics of SAR11-OTU in dmdA-detected GW waters suggest that water mass mixing plays a major role in shaping bacterial communities involved in phytoplankton-derived DMSP demethylation. C1 [Han, Dukki; Unno, Tatsuya] Jeju Natl Univ, Jeju Special Self Governing Prov, Jeju 63243, South Korea. [Kang, Hee Yoon; Kang, Chang-Keun; Hur, Hor-Gil] Gwangju Inst Sci & Technol, Sch Earth Sci & Environm Engn, Gwangju 61005, South Korea. RP Hur, HG (corresponding author), Gwangju Inst Sci & Technol, Sch Earth Sci & Environm Engn, Gwangju 61005, South Korea. 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Zeder, Michael Ameryk, Anetta Pernthaler, Jakob TI Seasonal dynamics and activity of typical freshwater bacteria in brackish waters of the Gulf of Gdansk SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB While typical freshwater and marine bacteria have been found to co-occur in brackish habitats, it is unknown if they are active members of the local bacterial assemblages or if their presence is the result of passive transport only. We followed the seasonal dynamics of typical freshwater bacteria (R-BT lineage of Betaproteobacteria; Ac1 Actinobacteria; LD12 Alphaproteobacteria) and of marine SAR11 Alphaproteobacteria in the brackish water of the Gulf of Gdansk (southern Baltic Sea), and we assessed their incorporation of thymidine and leucine at three distinct environmental conditions. The temporal development of bacteria was driven not only by local conditions but also by phenomena resulting from the dynamic hydrology of the site. 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In contrast, there was a decrease in the dominant SAR11 phylotype and an increase in Pseudoalteromonas spp. in the oiled mesocosms (investigated by 16S rRNA gene analysis using denaturing gradient gel electrophoresis), as a consequence of the longer incubation, closer proximity of the samples to oil, and the lack of replenishment with seawater. A total of 216 strains were isolated from hydrocarbon enrichment cultures, predominantly belonging to the genus Pseudoaltero monas; most strains grew on PAHs, branched and straight-chain alkanes, as well as many other carbon sources. No obligate hydrocarbonoclastic bacteria were isolated or detected, highlighting the potential importance of cosmopolitan marine generalists like Pseudoalteromonas spp. in degrading hydrocarbons in the water column beneath an oil slick, and revealing the susceptibility to oil pollution of SAR11, the most abundant bacterial clade in the surface ocean. 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Fuchsman, Clara A. TI Niche differentiation in microorganisms capable of using alternative reduced nitrogen sources studied across depth and between oxic and anoxic ocean regions SO FRONTIERS IN MARINE SCIENCE DT Article AB Introduction: Assimilation of reduced nitrogen is less energetically costly than assimilation of oxidized forms. In the open ocean, ammonium is generally absent from the water column, including in oxygen-deficient zones (ODZs). Some microorganisms can use alternative organic reduced nitrogen forms like urea and cyanate, as indicated by the presence of cyanase (cynS) and urease (ureC) genes. Methods: Here we examine the Hawaii Ocean Time series, two stations in the Eastern Tropical South Pacific ODZ and one in the Eastern Tropical North Pacific ODZ, using phylogenetic read placement of metagenomic reads to define the proportion of each taxon capable of using cyanate and/or urea in oxic and anoxic environments. Results: An improved phylogenetic tree found that Thioglobaceae and Verrucomicrobia had the capability to use urea. Our detailed examination of all the microbial groups able to use cyanate and urea illuminated that niche differentiation, an adaptation to minimize competition, determines chosen nitrogen sources, partitioning by depth and oxygen. Urease genes were found in Picocyanobacteria and SAR11 in surface waters, Thaumarchaeota and Nitrospina in deep waters, Thioglobaceae and Cand. Scalindua in ODZs, and Verrucomicrobia in the deep oxycline. In the ODZs, the percentage of Anammox bacteria that contained cynS was double that of those containing ureC, and their cynS transcripts were abundant, indicating a preference for cyanate over urea. Discussion: While Prochlorococcus could utilize cyanate in the deep chlorophyll maximum, in the ODZs, Prochlorococcus uses nitrite rather than compete with Cand. Scalindua for cyanate, even though cyanate is present. SAR11 and Prochlorococcus may compete for urea in surface waters, but for SAR11, the presence of ureC was negatively correlated with nitrate concentration (p = 10(-17)), with similar to 40% of SAR11 genomes containing the ureC gene in oxic surface waters but none at depth, indicating that SAR11 bacteria switched to using nitrate when available. In the oxycline above the ODZ, where Thaumarchaeota and Nitrospina both could use urea, 50% of Nitrospina were also able to use cyanate, and their cyanase transcripts were present. This use of dissolved organic N should allow a higher biomass of N-cycling microbes and higher N-transformation rates than in a system competing for ammonia only. C1 [Huanca-Valenzuela, Paulina; Cram, Jacob A.; Fuchsman, Clara A.] Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD 21613 USA. RP Fuchsman, CA (corresponding author), Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD 21613 USA. 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However, our current understanding of the correlation between bacterial genome size and environmental adaptation relies on too few species. It is still unclear whether there are other paths leading to genomic reduction in free-living bacteria. The genome size of marine free-living bacteria of the genus Idiomarina belonging to the order Alteromonadales (Gammaproteobacteria) is much smaller than the size of related genomes from bacteria in the same order. Comparative genomic and physiological analyses showed that the genomic reduction pattern in this genus is different from that of the classical SAR11 lineage. Genomic reduction reconstruction and substrate utilization profile showed that Idiomarina spp. lost a large number of genes related to carbohydrate utilization, and instead they specialized on using proteinaceous resources. Here we propose a new hypothesis to explain genomic reduction in this genus; we propose that trophic specialization increasing the metabolic efficiency for using one kind of substrate but reducing the substrate utilization spectrum could result in bacterial genomic reduction, which would be not uncommon in nature. This hypothesis was further tested in another free-living genus, Kangiella, which also shows dramatic genomic reduction. These findings highlight that trophic specialization is potentially an important path leading to genomic reduction in some marine free-living bacteria, which is distinct from the classical lineages like SAR11. IMPORTANCE The streamlining hypothesis is usually used to explain the genomic reduction events in free-living bacteria like SAR11. However, we find that the genomic reduction phenomenon in the bacterial genus Idiomarina is different from that in SAR11. Therefore, we propose a new hypothesis to explain genomic reduction in this genus based on trophic specialization that could result in genomic reduction, which would be not uncommon in nature. Not only can the trophic specialization hypothesis explain the genomic reduction in the genus Idiomarina, but it also sheds new light on our understanding of the genomic reduction processes in other free-living bacterial lineages. C1 [Qin, Qi-Long; Li, Yi; Sun, Lin-Lin; Wang, Zhi-Bin; Chen, Xiu-Lan; Zhang, Yu-Zhong] Shandong Univ, Marine Biotechnol Res Ctr, State Key Lab Microbial Technol, Qingdao, Peoples R China. [Wang, Shi; Zhang, Yu-Zhong] Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China. [Chen, Xiu-Lan; Zhang, Yu-Zhong] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China. [Oren, Aharon] Hebrew Univ Jerusalem, Alexander Silberman Inst Life Sci, Dept Plant & Environm Sci, Edmond J Safra Campus, Jerusalem, Israel. RP Zhang, YZ (corresponding author), Shandong Univ, Marine Biotechnol Res Ctr, State Key Lab Microbial Technol, Qingdao, Peoples R China.; Zhang, YZ (corresponding author), Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China.; Zhang, YZ (corresponding author), Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China.; Oren, A (corresponding author), Hebrew Univ Jerusalem, Alexander Silberman Inst Life Sci, Dept Plant & Environm Sci, Edmond J Safra Campus, Jerusalem, Israel. 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Martin, Adrian P. Scanlan, David J. Zubkov, Mikhail V. TI Photoheterotrophy of bacterioplankton is ubiquitous in the surface oligotrophic ocean SO PROGRESS IN OCEANOGRAPHY DT Review AB Accurate measurements in the Southern Hemisphere were obtained to test a hypothesis of the ubiquity of photoheterotrophy in the oligotrophic ocean. We present experimental results of light-enhanced uptake of methionine, leucine and ATP by bacterioplankton during two large-scale transects of the South Atlantic. Light increased the uptake of substrates by both dominant bacterioplankton groups, Prochlorococcus and SAR11, as well as for the bulk microbial community. Our consistent experimental evidence strongly indicates that photoheterotrophy is characteristic of dominant bacterioplankton populations in the global oligotrophic ocean. (C) 2015 Elsevier Ltd. All rights reserved. C1 [Evans, Claire; Gomez-Pereira, Paola R.; Martin, Adrian P.; Zubkov, Mikhail V.] Natl Oceanog Ctr, Ocean Biogeochem & Ecosyst Res Grp, Southampton, Hants, England. [Scanlan, David J.] Univ Warwick, Sch Life Sci, Coventry CV4 7AL, W Midlands, England. RP Zubkov, MV (corresponding author), Natl Oceanog Ctr, Ocean Biogeochem & Ecosyst Res Grp, European Way, Southampton, Hants, England. 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Bolanos, Luis M. Bell, Ashley G. Michelsen, Michelle L. Allen, Michael J. Temperton, Ben TI A Novel and Ubiquitous Marine Methylophage Provides Insights into Viral-Host Coevolution and Possible Host-Range Expansion in Streamlined Marine Heterotrophic Bacteria SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Isolation and cultivation of viruses are the foundations on which the mechanistic understanding of virus-host interactions and parameterization of bioinformatic tools for viral ecology are based. This study isolated and characterized the first myophage known to infect the OM43 clade, expanding our knowledge of this understudied group of microbes. The methylotrophic OM43 clade are Gammaproteobacteria that comprise some of the smallest free-living cells known and have highly streamlined genomes. OM43 represents an important microbial link between marine primary production and remineralization of carbon back to the atmosphere. Bacteriophages shape microbial communities and are major drivers of mortality and global marine biogeochemistry. Recent cultivation efforts have brought the first viruses infecting members of the OM43 clade into culture. Here, we characterize a novel myophage infecting OM43 called Melnitz. Melnitz was isolated independently from water samples from a subtropical ocean gyre (Sargasso Sea) and temperate coastal (Western English Channel) systems. Metagenomic recruitment from global ocean viromes confirmed that Melnitz is globally ubiquitous, congruent with patterns of host abundance. Bacteria with streamlined genomes such as OM43 and the globally dominant SAR11 clade use riboswitches as an efficient method to regulate metabolism. Melnitz encodes a two-piece tmRNA (ssrA), controlled by a glutamine riboswitch, providing evidence that riboswitch use also occurs for regulation during phage infection of streamlined heterotrophs. Virally encoded tRNAs and ssrA found in Melnitz were phylogenetically more closely related to those found within the alphaproteobacterial SAR11 clade and their associated myophages than those within their gammaproteobacterial hosts. This suggests the possibility of an ancestral host transition event between SAR11 and OM43. Melnitz and a related myophage that infects SAR11 were unable to infect hosts of the SAR11 and OM43, respectively, suggesting host transition rather than a broadening of host range. IMPORTANCE Isolation and cultivation of viruses are the foundations on which the mechanistic understanding of virus-host interactions and parameterization of bioinformatic tools for viral ecology are based. This study isolated and characterized the first myophage known to infect the OM43 clade, expanding our knowledge of this understudied group of microbes. The nearly identical genomes of four strains of Melnitz isolated from different marine provinces and the global abundance estimations from metagenomic data suggest that this viral population is globally ubiquitous. Genome analysis revealed several unusual features in Melnitz and related genomes recovered from viromes, such as a curli operon and virally encoded tmRNA controlled by a glutamine riboswitch, neither of which are found in the host. Further phylogenetic analysis of shared genes indicates that this group of viruses infecting the gammaproteobacterial OM43 shares a recent common ancestor with viruses infecting the abundant alphaproteobacterial SAR11 clade. Host ranges are affected by compatible cell surface receptors, successful circumvention of superinfection exclusion systems, and the presence of required accessory proteins, which typically limits phages to singular narrow groups of closely related bacterial hosts. This study provides intriguing evidence that for streamlined heterotrophic bacteria, virus-host transitioning may not be necessarily restricted to phylogenetically related hosts but is a function of shared physical and biochemical properties of the cell. C1 [Buchholz, Holger H.; Bolanos, Luis M.; Bell, Ashley G.; Michelsen, Michelle L.; Allen, Michael J.; Temperton, Ben] Univ Exeter, Sch Biosci, Exeter, Devon, England. [Allen, Michael J.] Plymouth Marine Lab, Plymouth, Devon, England. RP Temperton, B (corresponding author), Univ Exeter, Sch Biosci, Exeter, Devon, England. 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TI Prevalence of a calcium-based alkaline phosphatase associated with the marine cyanobacterium Prochlorococcus and other ocean bacteria SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>Phosphate plays a key role in regulating primary productivity in several regions of the world's oceans and here dissolved organic phosphate can be an important phosphate source. A key enzyme for utilizing dissolved organic phosphate is alkaline phosphatase and the phoA-type of this enzyme has a zinc cofactor. As the dissolved zinc concentration is low in phosphate depleted environments, this has led to the hypothesis that some phytoplankton may be zinc-P co-limited. Recently, it was shown that many marine bacteria contain an alternative form of alkaline phosphatase called phoX, but it is unclear which marine lineages carry this enzyme. Here, we describe the occurrence in low phosphate environments of phoX that is associated with uncultured Prochlorococcus and SAR11 cells. Through heterologous expression, we demonstrate that phoX encodes an active phosphatase with a calcium cofactor. The enzyme also functions with magnesium and copper, whereas cobalt, manganese, nickel and zinc inhibit enzyme activity to various degrees. We also find that uncultured SAR11 cells and cyanophages contain a different alkaline phosphatase related to a variant present in several Prochlorococcus isolates. Overall, the results suggest that many bacterial lineages including Prochlorococcus and SAR11 may not be subject to zinc-P co-limitation. C1 [Kathuria, Satish; Martiny, Adam C.] Univ Calif Irvine, Dept Earth Syst Sci, Irvine, CA 92697 USA. [Martiny, Adam C.] Univ Calif Irvine, Dept Ecol & Evolutionary Biol, Irvine, CA 92697 USA. RP Martiny, AC (corresponding author), Univ Calif Irvine, Dept Earth Syst Sci, Irvine, CA 92697 USA. 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Batailler, Nicole Caparros, Jocelyne Catala, Philippe Lantoine, Francois Oriol, Louise Lebaron, Philippe Obernosterer, Ingrid TI Impact of lower salinity waters on bacterial heterotrophic production and community structure in the offshore NW Mediterranean Sea SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB P>We investigated the impact of water masses originating from freshwater input on bacterial heterotrophic metabolism and community structure at an offshore site in the oligotrophic NW Mediterranean Sea in 2007 and 2008. By combining 16S rRNA gene clone libraries and MICRO-CARD-FISH we determined the dominant operational taxonomic units (OTU) and their contribution to bulk abundance and activity in the presence of buoyant water masses characterized by lower salinity (LSW, < 37.9) and compared these with the winter and spring phytoplankton blooms. 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The highest abundance of dmdA genes was in May 2008 at 25 m, with similar to 16.5% of cells harboring a gene in one of the eight subclades surveyed, while the highest abundance of dddP genes was in July 2008 at 25 m, with similar to 2% of cells harboring a gene. The dmdA gene pool was consistently dominated by homologs from SAR11 subclades, which was supported by findings in metagenomic data sets derived from Station ALOHA. Expression of the SAR11 dmdA genes was low, with typical transcript: gene ratios between 1:350 and 1:1,400. The abundance of DMSP genes was statistically different between 25 m and the DCM and correlated with a number of environmental variables, including primary production, photosynthetically active radiation, particulate DMSP, and DMS concentrations. At 25 m, dddP abundance was positively correlated with pigments that are diagnostic of diatoms; at the DCM, dmdA abundance was positively correlated with temperature. 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Halloush, Shiraz Al-jedai, Ahmed Alrashed, Ahmed AlFaifi, Mashael Mohzari, Yahya Almutairi, Malak AlHassar, Fatimah Howaidi, Jude Almutairi, Wedad Abraham, Ivo Alkhatib, Nimer TI Cost-effectiveness of Favipiravir in moderately to severely ill COVID-19 patients in the real-world setting of Saudi arabian pandemic referral hospitals SO SAUDI PHARMACEUTICAL JOURNAL DT Article AB Purpose: We aimed to evaluate the cost effectiveness of Favipiravir treatment versus standard of care (SC) in moderately to severely ill COVID-19 patients from the Saudi healthcare payer perspective. Methods: We used the patient-level simulation method to simulate a cohort of 415 patients with mod-erate to severe COVID-19 disease who were admitted to two Saudi COVID-19 referral hospitals: 220 patients on Favipiravir and 195 patients on SC. We estimated the incremental cost-effectiveness ratio (ICER) of Favipiravir versus SC in terms of the probability to be discharged alive from hospital and the mean time in days to discharge one patient alive. The model was performed twice: first, using unweighted, and second, using weighted clinical and economic data. Weighting using the inverse weight probability method was performed to achieve balance in baseline characteristics. Results: In the unweighted model, base case (probabilistic) ICER estimates favored Favipiravir at savings of Saudi Riyal (SAR)1,611,511 (SAR1,998,948) per 1% increase in the probability of being discharged alive. As to mean time to discharging one patient alive, ICERs favored Favipiravir at savings of SAR11,498 (SAR11,125). Similar results were observed in the weighted model with savings using Favipiravir of SAR1,514,893 (SAR2,453,551) per 1% increase in the probability of being discharged alive, and savings of SAR11,989 (SAR11,277) for each day a patient is discharged alive. Conclusion: From the payer perspective, the addition of Favipiravir in moderately to severely ill COVID-19 patients was cost-savings over SC. Favipiravir was associated with a higher probability of discharging patients alive and lower daily spending on hospitalization than SC.(c) 2023 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). C1 [Alamer, Ahmad] Prince Sattam Bin Abdulaziz Univ, Coll Pharm, Dept Clin Pharm, Alkharj, Saudi Arabia. [Alamer, Ahmad; Abraham, Ivo] Univ Arizona, Ctr Hlth Outcomes & Pharmacoecon Res, Tucson, AZ USA. [Almutairi, Abdulaali R.] Saudi Food & Drug Author, Drug Sect, Riyadh, Saudi Arabia. [Halloush, Shiraz] Appl Sci Private Univ, Fac Pharm, Dept Clin Pharm & Therapeut, Amman, Jordan. [Al-jedai, Ahmed] Deputyship Therapeut Affairs, Minist Hlth, Riyadh, Saudi Arabia. [Al-jedai, Ahmed] Al Faisal Univ, Coll Pharm, Riyadh, Saudi Arabia. [Alrashed, Ahmed; Howaidi, Jude] King Fahad Med City, Dept Clin Pharm, Riyadh, Saudi Arabia. [AlFaifi, Mashael; Mohzari, Yahya] King Fahad Med City, Riyadh, Saudi Arabia. [Almutairi, Malak] Almaarefa Univ, Coll Pharm, Dept Pharm Practice, Riyadh, Saudi Arabia. [AlHassar, Fatimah; Almutairi, Wedad] Imam Abdulrahman Bin Faisal Univ, Dept Clin Pharm, Dammam, Saudi Arabia. [Alkhatib, Nimer] Al Zaytoonah Univ Jordan, Fac Pharm, Dept Pharm, Amman, Jordan. [Alkhatib, Nimer] PI Pharm Intelligence, Amman, Jordan. RP Almutairi, AR (corresponding author), Saudi Food & Drug Author, Drug Sect, Riyadh, Saudi Arabia. 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Amann, Rudolf Zubkov, Mikhail V. Tarran, Glen A. Pernthaler, Jakob TI Latitudinal distribution of prokaryotic picoplankton populations in the Atlantic Ocean SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>Members of the prokaryotic picoplankton are the main drivers of the biogeochemical cycles over large areas of the world's oceans. In order to ascertain changes in picoplankton composition in the euphotic and twilight zones at an ocean basin scale we determined the distribution of 11 marine bacterial and archaeal phyla in three different water layers along a transect across the Atlantic Ocean from South Africa (32.9 degrees S) to the UK (46.4 degrees N) during boreal spring. Depth profiles down to 500 m at 65 stations were analysed by catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and automated epifluorescence microscopy. 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The fractions of SAR11, Polaribacter, and Ant4D3 that were active differed from each other and varied among substrates. SAR11 had the largest fraction of active cells incorporating leucine, while Polaribacter dominated the community using protein. The fraction of Ant4D3 using different compounds did not vary, but this group dominated the incorporation of amino acids, and was an abundant and active component of the bacterial community. Bacteria in the WAP region were as active as bacteria in the Mid-Atlantic Bight, even though total bacterial production was lower in the WAP. Though persistently cold (0-1 degrees C) and dominated by different bacterial taxa, the single-cell activity of this summertime Antarctic bacterial community was comparable to that of temperate communities. C1 [Straza, Tiffany R. A.; Kirchman, David L.] Univ Delaware, Sch Marine Sci & Policy, Lewes, DE 19958 USA. [Ducklow, Hugh W.] Marine Biol Lab, Ctr Ecosyst, Woods Hole, MA 02543 USA. [Murray, Alison E.] Univ Nevada, Desert Res Inst, Reno, NV 89506 USA. RP Kirchman, DL (corresponding author), Univ Delaware, Sch Marine Sci & Policy, Lewes, DE 19958 USA. 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Simo, Rafel TI Sunlight effects on the DMSP-sulfur and leucine assimilation activities of polar heterotrophic bacterioplankton SO BIOGEOCHEMISTRY DT Article; Proceedings Paper CT 5th International Symposium on Biological and Environmental Chemistry of DMS(P) and Related Compounds CY OCT 19-22, 2010 CL Natl Inst Oceanography (NIO), INDIA HO Natl Inst Oceanography (NIO) AB The influence of solar ultraviolet radiation and photosynthetically active radiation (PAR) on summertime marine bacterial uptake and assimilation of sulfur from radiolabeled dimethlysulfoniopropionate (S-35-DMSP) was studied at four Arctic and two Antarctic stations. Incubations with H-3-leucine were also conducted for comparative purposes as a measurement of bacterial activity. Arctic waters were characterized by large numbers of colonial Phaeocystis pouchetii and higher DMSP concentrations than in the two diatom-dominated Antarctic samples. Exposure to full sunlight radiation (280-700 nm), and to a lesser extent to PAR + UVA (320-700 nm), generally decreased the bacterial assimilation of H-3-leucine with respect to darkness, and caused variable effects on S-35-DMSP assimilation. By using a single-cell approach involving microautoradiography we found high percentages of sulfur assimilating cells within the bacterial groups Gammaproteobacteria, Bacteroidetes, SAR11 and Roseobacter despite the varying DMSP concentrations between Arctic and Antarctic samples. The dominant SAR11 clade contributed 50-70% of the cells assimilating both substrates in the Arctic stations, whereas either Gammaproteobacteria or SAR11 were the largest contributors to H-3-leucine uptake in samples from the two Antarctic stations. Only one station was analyzed for single-cell S-35-DMSP assimilation in Antarctica, and Gammaproteobacteria were major contributors to its uptake, providing the first evidence for Antarctic bacteria actively taking up S-35-DMSP. PAR + UVA repeatedly increased the number of SAR11 cells assimilating H-3-leucine. This pattern also occurred with other S-35-DMSP assimilating groups, though not so consistently. Our results support a widespread capability of polar bacteria to assimilate DMSP-sulfur during the season of maximum DMSP concentrations, and show for the first time that all major polar taxa can be highly active at this assimilation under the appropriate circumstances. Our findings further confirm the role of sunlight as a modulator of heterotrophic carbon and sulfur fluxes in the surface ocean. C1 [Ruiz-Gonzalez, Clara; Gali, Marti; Gasol, Josep M.; Simo, Rafel] CSIC, Inst Ciencies Mar, E-08003 Barcelona, Catalunya, Spain. 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Microbial methanol dissimilation varied between 0.05 and 1.68 nmol L-1 h(-1) in the top 200 m of the Atlantic Ocean and showed significant variability between biogeochemical provinces. The highest rates of methanol dissimilation were found in the northern subtropical gyre (average 0.99 +/- 0.41 nmol L-1 h(-1)), which were up to 8 times greater than other Atlantic regions. Microbial methanol dissimilation rates displayed a significant inverse correlation with heterotrophic bacterial production (determined using H-3-leucine). Despite significant depth stratification of bacterial communities, methanol dissimilation rates showed much greater variability between oceanic provinces compared to depth. There were no significant differences in rates between samples collected under light and dark environmental conditions. The variability in the numbers of SAR11 (16S rRNA gene sequences) were estimated to explain approximately 50 % of the changes in microbial methanol dissimilation rates. 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To address this problem, we used a water circulation culture system that provided in-water air bubbles to achieve active growth and mass cultivation of the dinoflagellate Alexandrium affine. This facilitated assessment in a large-scale mesocosm (1000 L) of the short-term response of the marine pelagic planktonic community to inoculation with A. affine under diatom-dominant conditions. Members of the pelagic planktonic community (bacteria and phytoplankton) varied in their response to A. affine addition, even over short time periods. Following nutrient depletion, the population of the dominant diatom (Chaetoceros debilis) rapidly declined, while the proportion of A. affine increased to more than 80% in both the low and high concentration treatment groups. The bacterial community was influenced by the population dynamics of phytoplankton, but a similar effect was not evident for the mesozooplankton community. Rhodobacteraceae dominated during the diatom-dominant stage in the mesocosms, and their numbers were positively correlated with most of the diatoms, including C. debilis. In contrast, the bacterial SAR11 clade and C. debilis showed markedly opposite patterns, and their occurrences were strongly negatively correlated. At the end of the experiment, there were similar time-series changes evident between the major bacterial group, and phytoplankton of the SAR11 clade and A. affine, but these groups were not directly correlated in association network analysis, indicating that the SAR11 clade was more associated with the decline in diatom populations rather than having a stimulatory effect on dinoflagellates. 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Sachdeva, Rohan Cram, Jacob A. Steele, Joshua A. Needham, David M. Patel, Anand Parada, Alma E. Fuhrman, Jed A. TI Temporal variability and coherence of euphotic zone bacterial communities over a decade in the Southern California Bight SO ISME JOURNAL DT Article AB Time-series are critical to understanding long-term natural variability in the oceans. Bacterial communities in the euphotic zone were investigated for over a decade at the San Pedro Ocean Time-series station (SPOT) off southern California. Community composition was assessed by Automated Ribosomal Intergenic Spacer Analysis (ARISA) and coupled with measurements of oceanographic parameters for the surface ocean (0-5 m) and deep chlorophyll maximum (DCM, average depth similar to 30 m). SAR11 and cyanobacterial ecotypes comprised typically more than one-third of the measured community; diversity within both was temporally variable, although a few operational taxonomic units (OTUs) were consistently more abundant. 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R. Juergens, Klaus TI Zonation of bacterioplankton communities along aging upwelled water in the northern Benguela upwelling SO FRONTIERS IN MICROBIOLOGY DT Article AB Upwelling areas are shaped by enhanced primary production in surface waters, accompanied by a well-investigated planktonic succession. Although bacteria play an important role in biogeochemical cycles of upwelling systems, little is known about bacterial community composition and its development during upwelling events. The aim of this study was to investigate the succession of bacterial assemblages in aging upwelled water of the Benguela upwelling from coastal to offshore sites. Water from the upper mixed layer at 12 stations was sampled along two transects from the origin of the upwelling to a distance of 220 km. 16S rRNA gene amplicon sequencing was then used in a bacterial diversity analysis and major bacterial taxa were quantified by catalyzed reporter deposition-fluorescence in situ hybridization. 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The sequencing effort was directed by denaturing gradient gel electrophoresis (DGGE) screening of samples and the clone libraries. Sequences of 88 clones fell into seven major lineages of the domain Bacteria: alpha (36%)-, gamma (32%)-, delta (14%)-, and epsilon (1%)-Proteobacteria; Cytophaga-Flexibacter-Bacteroides spp. (9%); Verrucomicrobium spp. (6%); and green nonsulfur bacteria (2%). A total of 34% of the cloned sequences (excluding clones in the SAR11 and Roseobacter groups) had sequence similarities that were <94% compared to previously reported sequences, indicating the presence of novel sequences. DGGE fingerprints of the selected samples showed that most of the bands were common to all samples in all three seasons. However, additional bands representing sequences related to Cytophaga and Polaribacter species were found in samples collected during the summer and fall. Of the clones in a library generated from one sample collected in spring of 1995, 50% were the same and were most closely affiliated (99% similarity) with Alteromonas macleodii, while 50% of the clones in another sample were most closely affiliated (90 to 96% similarity) with Oceanospirillum sp. The majority of the cloned sequences were most closely related to uncultured, environmental sequences. Prominent among these were members of the SAR11 group. Differences between mixed-layer and halocline samples were apparent in DGGE fingerprints and clone libraries. Sequences related to alpha-Proteobacteria (dominated by SAR11) were abundant (52%) in samples from the mixed layer, while sequences related to gamma-proteobacteria were more abundant (44%) in halocline samples. Two bands corresponding to sequences related to SAR307 (common in deep water) and the high-G+C gram-positive bacteria were characteristic of the halocline samples. C1 Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. 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Wang, Shuting Wang, Kaiceng Chang, Kevin Li, Lanjuan TI Airborne polystyrene microplastics and nanoplastics induce nasal and lung microbial dysbiosis in mice SO CHEMOSPHERE DT Article AB Microplastics (MP) and nanoplastics (NP) have been found in multiple environments and creatures. However, their effects on the airway microbiota still remain poorly understood. In this study, a series of bioinformatic and statistical analyses were carried out to explore the influence of airborne MP and NP on the nasal and lung microbiota in mice. Both MP and NP were capable of inducing nasal microbial dysbiosis, and MP had a stronger influence on the lung microbiota than NP. Multiple nasal and lung bacteria were associated with MP and NP groups, among which nasal Staphylococcus and lung Roseburia were most associated with MP group, while nasal Prevotella and lung unclassified_Muribaculaceae were most associated with NP group. The nasal Staphylococcus, lung Roseburia, lung Eggerthella and lung Corynebacterium were associated with both MP and NP groups, which were potential biomarkers of micro/nanoplastics-induced airway dysbiosis. SAR11_Clade_Ia and SAR11_Clade_II were associated with both nasal and lung microbiota in MP group, while no such bacterium was determined in NP group. 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Santos, Thays T. L. TI Spatial and temporal patterns in the Pelagibacteraceae across an estuarine gradient SO FEMS MICROBIOLOGY ECOLOGY DT Article AB Marine bacterial communities show strong spatial and seasonal patterns, often characterized by changes at high taxonomic levels. The Pelagibacteraceae are common members of bacterial communities, with well-documented biogeography at the subclade level. To identify patterns within the subclades, the abundance and diversity of Pelagibacteraceae were analyzed over a two-year period at four stations across an estuarine gradient. Pelagibacteraceae was the most abundant bacterial family, averaging 27% of the community, but varying from 1% to 57% in any one sample. Highest abundances were detected in autumn and winter. Pelagibacteraceae richness was lowest at the most inshore site, and highest in autumn and winter at all sites. Shannon diversity decreased in winter, when a few OTUs dominated the community. Dissolved oxygen, dissolved silicate and prokaryote abundance explained most of the variability in the Pelagibacteraceae communities, with salinity differentiating low salinity communities. The 10 most abundant OTUs included OTUs that varied across sites, with little seasonality as well as those with small site effects, but strong seasonal patterns indicating differences in the niches of individual OTUs. While salinity was important in structuring low salinity communities, higher salinity communities appear to be responding to additional environmental parameters including oxygen, nutrients and other organisms.Closely related Pelagibacteraceae OTUs vary spatially and temporally across an estuary due to differences in salinity as well as oxygen, nutrients and the abundance of other organisms.Closely related Pelagibacteraceae OTUs vary spatially and temporally across an estuary due to differences in salinity as well as oxygen, nutrients and the abundance of other organisms. C1 [Ortmann, Alice C.] Univ S Alabama, Dept Marine Sci, Mobile, AL 36688 USA. [Ortmann, Alice C.] Dauphin Isl Sea Lab, Dauphin Isl, AL 36528 USA. [Santos, Thays T. L.] Univ Fed Maranhao, Dept Oceanog & Limnol, BR-65080805 Sao Luis, MA, Brazil. [Santos, Thays T. L.] Univ Maine, Sch Marine Sci, Orono, ME 04469 USA. RP Ortmann, AC (corresponding author), Bedford Inst Oceanog, Ctr Offshore Oil Gas & Energy Res, Dept Fisheries & Ocean Canada, Dartmouth, NS B2Y 4A2, Canada. 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TI Variational inference for microbiome survey data with application to global ocean data SO ISME COMMUNICATIONS DT Article AB Linking sequence-derived microbial taxa abundances to host (patho-)physiology or habitat characteristics in a reproducible and interpretable manner has remained a formidable challenge for the analysis of microbiome survey data. Here, we introduce a flexible probabilistic modeling framework, VI-MIDAS (variational inference for microbiome survey data analysis), that enables joint estimation of context-dependent drivers and broad patterns of associations of microbial taxon abundances from microbiome survey data. VI-MIDAS comprises mechanisms for direct coupling of taxon abundances with covariates and taxa-specific latent coupling, which can incorporate spatio-temporal information and taxon-taxon interactions. We leverage mean-field variational inference for posterior VI-MIDAS model parameter estimation and illustrate model building and analysis using Tara Ocean Expedition survey data. Using VI-MIDAS' latent embedding model and tools from network analysis, we show that marine microbial communities can be broadly categorized into five modules, including SAR11-, nitrosopumilus-, and alteromondales-dominated communities, each associated with specific environmental and spatiotemporal signatures. VI-MIDAS also finds evidence for largely positive taxon-taxon associations in SAR11 or Rhodospirillales clades, and negative associations with Alteromonadales and Flavobacteriales classes. Our results indicate that VI-MIDAS provides a powerful integrative statistical analysis framework for discovering broad patterns of associations between microbial taxa and context-specific covariate data from microbiome survey data. C1 [Mishra, Aditya] Univ Georgia, Dept Stat, Athens, GA 30606 USA. 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Kirkpatrick, John B. Brazelton, William J. Murray, James W. Staley, James T. TI Metabolic strategies of free-living and aggregate-associated bacterial communities inferred from biologic and chemical profiles in the Black Sea suboxic zone SO FEMS MICROBIOLOGY ECOLOGY DT Article AB The Black Sea is a permanently anoxic basin with a well-defined redox gradient. We combine environmental 16S rRNA gene data from clone libraries, terminal restriction fragment length polymorphisms, and V6 hypervariable region pyrosequences to provide the most detailed bacterial survey to date. Furthermore, this data set is informed by comprehensive geochemical data; using this combination of information, we put forward testable hypotheses regarding possible metabolisms of uncultured bacteria from the Black Sea's suboxic zone (microaerophily, nitrate reduction, manganese cycling, and oxidation of methane, ammonium, and sulfide). Dominant bacteria in the upper suboxic zone included members of the SAR11, SAR324, and Microthrix groups and in the deep suboxic zone included members of BS-GSO-2, Marine Group A, and SUP05. A particulate fraction (30 mu m filter) was used to distinguish between free-living and aggregate-attached communities in the suboxic zone. The particulate fraction contained greater diversity of V6 tag sequences than the bulk water samples. Lentisphaera, Epsilonproteobacteria, WS3, Planctomycetes, and Deltaproteobacteria were enriched in the particulate fraction, whereas SAR11 relatives dominated the free-living fraction. On the basis of the bacterial assemblages and simple modeling, we find that in suboxic waters, the interior of sinking aggregates potentially support manganese reduction, sulfate reduction, and sulfur oxidation. C1 [Fuchsman, Clara A.; Kirkpatrick, John B.; Brazelton, William J.; Murray, James W.] Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. 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WOS:000297156300015 DA 2025-07-30 ER PT J AU Wei, B AF Wei, Bo TI Diversity and distribution of proteorhodopsin-containing microorganisms in marine environments SO FRONTIERS OF ENVIRONMENTAL SCIENCE & ENGINEERING DT Article AB Proteorhodopsin (PR) is a recently discovered protein involved in the utilization of light energy. Several studies have shown that PR-containing microorganisms are widespread and compose a large proportion of the biomass in marine ecosystems. A better understanding of the ecological role of PR will help clarify the effect of the global flow of energy and the carbon cycle on marine communities. In this study, a bioinformatical database of PR codon sequences, the Global Distribution Database of Proteorhodopsin (GDDP), as a tool for analyzing the diversity and distribution of PR-containing microorganisms in marine environments throughout the world was designed. The community structure of PR microorganisms were also compared using PCR assays and UniFrac analyses of 12 samples collected from three water layers (0, 75, and 200 m) at four representative sites in the Pacific, Atlantic, and Indian Oceans. The results indicate that PRcontaining microorganisms can be grouped into two distribution types: widespread and location-specific. 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Multiple studies have identified the significant influence of anoxic conditions (<= 1 mu M O-2) on marine prokaryotic communities and biogeochemical cycling of elements. However, little attention has been paid to the expanding low-oxygen zones where the oxygen level is still above the anoxic level. Here, we studied the abundance and taxonomic and functional profiles of prokaryotic communities in the Bay of Bengal (BoB), where the oxygen concentration is barely above suboxic level (5 mu M O-2). We found the sinking of Trichodesmium into deep water was far more efficient than that of Prochlorococcus, suggesting Trichodesmium blooms might be an essential carbon and nitrogen source for the maintenance of the BoB OMZ. In addition to the shift in the prokaryotic community composition, the abundance of some functional genes also changed with the change of oxygen concentration. Compared to oxic (>60 mu M O-2) Tara Ocean and high-hypoxic (>20 to <= 60 mu M O-2) BoB samples, we found more SAR11-nar sequences (responsible for reducing nitrate to nitrite) in low-hypoxic (>5 to <= 20 mu M O-2) BoB waters. This suggested SAR11-nar genes would be more widespread due to the expansion of OMZs. It seems that the nitrite-N was not further reduced to nitrogen through denitrification but likely oxidized to nitrate by Nitrospinae in the BoB OMZ and then accumulated in the form of nitrate-N. However, the lack of N-2 production in the BoB would change if the BoB OMZ became anoxic. Together, these results suggested that reduction of oxygen concentration and OMZ expansion may increase the use of nitrate by SAR11 and N-2 production in the BoB. IMPORTANCE Recognizing the prokaryotic community and its functions in hypoxic (>5 to <= 60 mu M O-2) environments before further expansion of OMZs is critical. We demonstrate the prokaryotic community and its potential functions in nitrogen metabolism in the Bay of Bengal (BoB), where oxygen concentration is barely above suboxic level. This study highlighted that Trichodesmium might be an essential carbon and nitrogen source in the maintenance of the BoB OMZ. Additionally, we suggest that the lack of N-2 production in the BoB would change if the BoB OMZ became anoxic, and the expansion of OMZs in the global ocean may potentially increase the use of nitrate by SAR11. Recognizing the prokaryotic community and its functions in hypoxic (>5 to <= 60 mu M O-2) environments before further expansion of OMZs is critical. We demonstrate the prokaryotic community and its potential functions in nitrogen metabolism in the Bay of Bengal (BoB), where oxygen concentration is barely above suboxic level. C1 [Gu, Bowei; Liu, Jiaxing; Tan, Yehui; Xia, Xiaomin] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou, Peoples R China. [Gu, Bowei; Liu, Jiaxing; Tan, Yehui; Xia, Xiaomin] Univ Chinese Acad Sci, Beijing, Peoples R China. [Gu, Bowei; Liu, Jiaxing; Tan, Yehui; Xia, Xiaomin] Southern Marine Sci & Engn Guangdong Lab Guangzh, Guangzhou, Peoples R China. [Cheung, Shunyan; Ho, Ngai Hei Ernest] Hong Kong Univ Sci & Technol, Dept Ocean Sci, Hong Kong, Peoples R China. [Cheung, Shunyan] Hong Kong Univ Sci & Technol, Southern Marine Sci & Engn Guangdong Lab, Hong Kong Branch, Hong Kong, Peoples R China. RP Xia, XM (corresponding author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou, Peoples R China.; Xia, XM (corresponding author), Univ Chinese Acad Sci, Beijing, Peoples R China.; Xia, XM (corresponding author), Southern Marine Sci & Engn Guangdong Lab Guangzh, Guangzhou, Peoples R China. 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In the future, the newly developed community analysis pipeline will allow for on-site insights into a microbial community within 35 h of sampling, which will permit more targeted sampling efforts and hypothesis-driven research. IMPORTANCE The South Pacific Gyre, due to its vast size and remoteness, is one of the least-studied oceanic regions on earth. However, both remote sensing and in situ measurements indicated that the activity of its microbial community contributes significantly to global biogeochemical cycles. Presented here is an unparalleled investigation of the microbial community of the SPG from 20- to 5,000-m depths covering a geographic distance of similar to 7,000 km. This insight was achieved through the development of a novel on-board analysis pipeline, which combines next-generation sequencing with fluorescence in situ hybridization and automated cell enumeration. 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A critical aspect of automated microbiome sampling is the preservation of nucleic acids over long-term autosampler deployments. Understanding the impact of preservation method on microbial metabarcoding is essential for implementing genomic observatories into existing infrastructure, and for establishing best practices for the regional and global synthesis of data. The present study evaluates the effect of two preservatives commonly used in autosampler deployments (mercuric chloride and formalin) and two extraction kits (PowerWater and NucleoSpin) on amplicon sequencing of 16S and 18S rRNA gene over 50 weeks of sample storage. Our results suggest the combination of mercuric chloride preservation and PowerWater extraction as most adequate for 16S and 18S rRNA gene amplicon-sequencing from the same seawater sample. This approach provides consistent information on species richness, diversity and community composition in comparison to control samples (nonfixed, filtered and frozen) when stored up to 50 weeks at in situ temperature. Preservation affects the recovery of certain taxa, with specific OTUs becoming overrepresented (SAR11 and diatoms) or underrepresented (Colwellia and pico-eukaryotes) after preservation. In case eukaryotic sequence information is the sole target, formalin preservation and NucleoSpin extraction performed best. Our study contributes to the design of long-term autonomous microbial observations in remote ocean areas, allowing cross-comparison of microbiome dynamics across sampling devices (e.g., water and particle samplers) and marine realms. C1 [Wietz, Matthias; Bienhold, Christina; Janssen, Felix; Salter, Ian; Boetius, Antje] Helmholtz Ctr Polar & Marine Res, Alfred Wegener Inst, Deep Sea Ecol & Technol, Bremerhaven, Germany. [Wietz, Matthias; Bienhold, Christina; Janssen, Felix; Boetius, Antje] Max Planck Inst Marine Microbiol, Bremen, Germany. [Metfies, Katja; Wolf, Christian] Helmholtz Ctr Polar & Marine Res, Polar Biol Oceanog, Alfred Wegener Inst, Bremerhaven, Germany. [Metfies, Katja] Carl von Ossietzky Univ Oldenburg, Helmholtz Inst Funct Marine Biodivers, Oldenburg, Germany. [Salter, Ian] Faroe Marine Res Inst, Torshavn, Faroe Islands. [Boetius, Antje] Univ Bremen, MARUM Ctr Marine Environm Sci, Bremen, Germany. RP Wietz, M (corresponding author), Helmholtz Ctr Polar & Marine Res, Alfred Wegener Inst, Deep Sea Ecol & Technol, Bremerhaven, Germany.; Wietz, M (corresponding author), Max Planck Inst Marine Microbiol, Bremen, Germany. 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We illustrate bacterial community shifts within the eddy centre, differentiating populations uplifted along isopycnals from those enriched or depleted at horizons of enhanced bacterial and primary productivity. Phylotypes belonging to the Roseobacter, OCS116 and marine Actinobacteria clades were enriched in the eddy core and were highly correlated with pigment-based indicators of diatom abundance, supporting developing hypotheses that members of these clades associate with phytoplankton blooms. Typical mesopelagic clades (SAR202, SAR324, SAR406 and SAR11 IIb) were uplifted within the eddy centre, increasing bacterial diversity in the lower euphotic zone. Typical surface oligotrophic clades (SAR116, OM75, Prochlorococcus and SAR11 Ia) were relatively depleted in the eddy centre. The biogeochemical context of a bloom-inducing eddy provides insight into the ecology of the diverse uncultured bacterioplankton dominating the oligotrophic oceans. 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Here, we report a first detailed bacterial inventory from vertical profiles of 60 sampling stations distributed along the salinity gradient of the Baltic Sea, one of world's largest brackish water environments, generated using 454 pyrosequencing of partial (400 bp) 16S rRNA genes. Within the salinity gradient, bacterial community composition altered at broad and finer-scale phylogenetic levels. Analogous to faunal communities within brackish conditions, we identified a bacterial brackish water community comprising a diverse combination of freshwater and marine groups, along with populations unique to this environment. As water residence times in the Baltic Sea exceed 3 years, the observed bacterial community cannot be the result of mixing of fresh water and saltwater, but our study represents the first detailed description of an autochthonous brackish microbiome. In contrast to the decline in the diversity of multicellular organisms, reduced bacterial diversity at brackish conditions could not be established. It is possible that the rapid adaptation rate of bacteria has enabled a variety of lineages to fill what for higher organisms remains a challenging and relatively unoccupied ecological niche. The ISME Journal (2011) 5, 1571-1579; doi: 10.1038/ismej.2011.41; published online 7 April 2011 C1 [Andersson, Anders F.] KTH Royal Inst Technol, Sci Life Lab, Sch Biotechnol, SE-10450 Stockholm, Sweden. [Herlemann, Daniel P. R.; Labrenz, Matthias; Juergens, Klaus; Waniek, Joanna J.] Leibniz Inst Baltic Sea Res, Dept Biol Oceanog, Rostock, Germany. [Bertilsson, Stefan] Uppsala Univ, Dept Ecol & Genet, Uppsala, Sweden. RP Andersson, AF (corresponding author), KTH Royal Inst Technol, Sci Life Lab, Sch Biotechnol, Box 24075, SE-10450 Stockholm, Sweden. 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Yu, Liying Heidelberg, John F. Kirchman, David L. TI Activity of abundant and rare bacteria in a coastal ocean SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB The surface layer of the oceans and other aquatic environments contains many bacteria that range in activity, from dormant cells to those with high rates of metabolism. However, little experimental evidence exists about the activity of specific bacterial taxa, especially rare ones. Here we explore the relationship between abundance and activity by documenting changes in abundance over time and by examining the ratio of 16S rRNA to rRNA genes (rDNA) of individual bacterial taxa. The V1-V2 region of 16S rRNA and rDNA was analyzed by tag pyrosequencing in a 3-y study of surface waters off the Delaware coast. Over half of the bacterial taxa actively cycled between abundant and rare, whereas about 12% always remained rare and potentially inactive. 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Chisholm, Sallie W. TI Metabolic evolution and the self-organization of ecosystems SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Metabolism mediates the flow of matter and energy through the biosphere. We examined how metabolic evolution shapes ecosystems by reconstructing it in the globally abundant oceanic phytoplankter Prochlorococcus. To understand what drove observed evolutionary patterns, we interpreted them in the context of its population dynamics, growth rate, and light adaptation, and the size and macromolecular and elemental composition of cells. This multilevel view suggests that, over the course of evolution, there was a steady increase in Prochlorococcus' metabolic rate and excretion of organic carbon. We derived a mathematical framework that suggests these adaptations lower the minimal subsistence nutrient concentration of cells, which results in a drawdown of nutrients in oceanic surface waters. This, in turn, increases total ecosystem biomass and promotes the coevolution of all cells in the ecosystem. Additional reconstructions suggest that Prochlorococcus and the dominant cooccurring heterotrophic bacterium SAR11 form a coevolved mutualism that maximizes their collective metabolic rate by recycling organic carbon through complementary excretion and uptake pathways. Moreover, the metabolic codependencies of Prochlorococcus and SAR11 are highly similar to those of chloroplasts and mitochondria within plant cells. These observations lead us to propose a general theory relating metabolic evolution to the self-amplification and self-organization of the biosphere. We discuss the implications of this framework for the evolution of Earth's biogeochemical cycles and the rise of atmospheric oxygen. C1 [Braakman, Rogier; Chisholm, Sallie W.] MIT, Dept Civil & Environm Engn, Cambridge, MA 02139 USA. [Braakman, Rogier; Follows, Michael J.] MIT, Dept Earth Atmospher & Planetary Sci, Cambridge, MA 02139 USA. [Chisholm, Sallie W.] MIT, Dept Biol, Cambridge, MA 02139 USA. RP Braakman, R; Chisholm, SW (corresponding author), MIT, Dept Civil & Environm Engn, Cambridge, MA 02139 USA.; Braakman, R (corresponding author), MIT, Dept Earth Atmospher & Planetary Sci, Cambridge, MA 02139 USA.; Chisholm, SW (corresponding author), MIT, Dept Biol, Cambridge, MA 02139 USA. 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Moreover, we studied the relationship between bacterial and small-sized eukaryotic community composition to identify potential biotic interactions among components of these two communities. Bacterial operational taxonomic unit (OTU) richness and diversity systematically peaked in autumn-winter, likely related to low resource availability during this period. BCC showed seasonal and vertical patterns, with Rhodobacteraceae and Flavobacteriaceae families dominating in surface waters, and SAR11 clade dominating at the base of the photic zone (30 m depth). BCC variability was significantly explained by environmental variables (e.g., temperature of water, solar radiation, or dissolved organic matter). Interestingly, a strong and significant correlation was found between BCC and small-sized eukaryotic community composition (ECC), which suggests that biotic interactions may play a major role as structuring factors of the microbial plankton in this productive area. 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The proportion of cells using starch (mean 56% 64%) or leucine (mean 47% 64%) was significantly higher than that using bicarbonate. On average, SAR11, Roseobacter spp. and Gammaproteobacteria showed a similarly high percentage of cells using leucine (47%-65% of hybridized cells) than using starch (51%-64% of hybridized cells), while Bacteroidetes and SAR86 cells preferentially used starch (53% of hybridized cells) over leucine (34%-40% of hybridized cells). We suggest that the great percentage of bacteria using starch is related to a high ambient availability of polymers associated to algal cell lysis, which, in turn, weakens the short-term coupling between phytoplankton release and bacterial production. C1 [Teira, E.; Hernando-Morales, V.] Univ Vigo, Dept Ecoloxia & Bioloxia Anim, Vigo 36310, Spain. [Teira, E.; Hernando-Morales, V.] Univ Vigo, Estn Ciencias Marinas Toralla ECIMAT, Vigo 36331, Spain. [Guerrero-Feijoo, E.; Varela, M. 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Ferrera, Isabel TI Effects of grazing, phosphorus and light on the growth rates of major bacterioplankton taxa in the coastal NW Mediterranean SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB Estimation of growth rates is crucial to understand the ecological role of prokaryotes and their contribution to marine biogeochemical cycling. However, there are only a few estimates for individual taxa. Two top-down (grazing) and bottom-up (phosphorus (P) availability) manipulation experiments were conducted under different light regimes in the NW Mediterranean Sea. Growth rate of different phylogenetic groups, including the Bacteroidetes, Rhodobacteraceae, SAR11, Gammaproteobacteria and its subgroups Alteromonadaceae and the NOR5/OM60 clade, were estimated from changes in cell numbers. Maximal growth rates were achieved in the P-amended treatments but when comparing values between treatments (response ratios), the response to predation removal was in general larger than to P-amendment. 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TI Distinct bacterial groups contribute to carbon cycling during a naturally iron fertilized phytoplankton bloom in the Southern Ocean SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB We investigated the contribution of distinct bacterial groups to bulk abundance and leucine incorporation during a spring phytoplankton bloom induced by natural iron fertilization in the Southern Ocean (Kerguelen Ocean and Plateau Compared Study, January-February 2005). Oligonucleotide probes were designed to target five operational taxonomic units (OTUs) at a narrow phylogenetic level (>= 99% identity of the 16S ribosomal ribonucleic acid [rRNA] gene). During the peak of the phytoplankton bloom, the Roseobacter groups NAC11-7 and RCA, the OTUs SAR92 belonging to Gammaproteobacteria, and the Bacteroidetes OTU Agg58 dominated bulk abundance and leucine incorporation. These four OTUs disappeared with the decline of the bloom, when the cosmopolitan groups SAR11 and SAR86 became dominant. 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Yoshizawa, Susumu Boeuf, Dominique Iwasaki, Wataru DeLong, Edward F. TI Proteorhodopsin variability and distribution in the North Pacific Subtropical Gyre SO ISME JOURNAL DT Article AB Proteorhodopsin is a light-activated retinal-containing proton pump found in many marine bacteria. These photoproteins are globally distributed in the ocean's photic zone and are capable of generating a proton motive force across the cell membrane. We investigated the phylogenetic diversity, distribution, and abundance of proteorhodopsin encoding genes in free-living bacterioplankton in the North Pacific Subtropical Gyre, leveraging a gene catalog derived from metagenomic samples from the ocean's surface to 1000 m depth. Proteorhodopsin genes were identified at all depths sampled, but were most abundant at depths shallower than 200 m. The majority of proteorhodopsin gene sequences (60.9%) belonged to members of the SAR11 lineage, with remaining sequences distributed among other diverse taxa. We observed variations in the conserved residues involved in ion pumping and spectral tuning, and biochemically confirmed four different proton pumping proteorhodopsin motifs, including one unique to deep-water SAR11. We also identified a new group of putative proteorhodopsins having unknown function. Our results reveal a broad organismal and unexpected depth distribution for different proteorhodopsin types, as well as substantial within-taxon variability. These data provide a framework for exploring the ecological relevance of proteorhodopsins and their spatiotemporal variation and function in heterotrophic bacteria in the open ocean. C1 [Olson, Daniel K.; Boeuf, Dominique; DeLong, Edward F.] Univ Hawaii, Dept Oceanog, Daniel K Inouye Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. [Yoshizawa, Susumu; Iwasaki, Wataru] Univ Tokyo, Atmosphere & Ocean Res Inst, Chiba 2778564, Japan. [Iwasaki, Wataru] Univ Tokyo, Grad Sch Sci, Dept Biol Sci, Tokyo 1130032, Japan. 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Heywood, Jane L. Ward, Peter Zubkov, Mikhail V. TI Bacterioplankton composition in the Scotia Sea, Antarctica, during the austral summer of 2003 SO AQUATIC MICROBIAL ECOLOGY DT Article AB Physical ocean processes (ice-melt, island run-off and upwelling of nutrients) were hypothesised to affect the bacterioplankton composition in the surface mixed layer of the Scotia Sea during the austral summer of 2003, and this was investigated using flow cytometry and catalysed reporter deposition fluorescence in situ hybridisation (CARD-FISH) techniques. The bacterioplankton was composed predominantly of Alphaproteobacteria (PB), comprising SAR11, Roseobacter spp. and SAR116 groups, followed by Sphingobacteria/Flavobacteria and Gammaproteobacteria, including SAR86. Two distinct bacterioplankton communities were identified, largely based on bacterioplankton abundance, which varied from 0.3 +/- 0.06 x 10(6) cells ml(-1) in the west to 0.8 +/- 0.3 x 10(6) cells ml(-1) in the east, and a corresponding difference in SAR11 percentages of 30 +/- 15% in the west compared to 5 +/- 5% in the east. The western community was present in waters that were largely in an over-wintered, pre-bloom condition. The eastern bacterioplankton community was associated with phytoplankton blooms developed within the eastern Scotia Sea nutrient upwelling zone, where the Antarctic Circumpolar Current (ACC) encounters the shallow bathymetry associated with the Scotia Arc, in combination with seasonal ice-melt and island effects that enabled surface water stratification. C1 Natl Oceanog Ctr, Southampton SO14 3ZH, Hants, England. British Antarctic Survey, NERC, Cambridge CB3 0ET, England. 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TI The minimal genome-a metabolic and environmental comparison SO BRIEFINGS IN FUNCTIONAL GENOMICS DT Article AB The field of Synthetic Biology seeks to apply engineering principles to biology in order to produce novel biological systems. One approach to accomplish this goal is the genome-driven cell engineering approach, which searches for functioning minimal genomes in naturally occurring microorganisms, which can then be used as a template for future systems. Currently a prototypical minimal genome has not been discovered. This review analyzes the organisms Mycoplasma pneumoniae, Pelagibacter ubique, Vesicomyosocius okutanii and Prochlorococcus marinus as models of heterotrophic symbiont, heterotrophic free-living, autotrophic symbiont and autotrophic free-living organisms respectively and compares them to the current minimal cell model in order to determine which most closely resembles a true minimal genome. M. pneumoniae possesses a genome of 816 394 base pairs (bp) with 688 open reading frames (ORF) and a severely limited metabolism. Pelagibacter ubique possesses a 1 308 000 bp genome with 1354 ORF and has a fully functional metabolism but requires a reduced form of sulphur. Vesicomyosocius okutanii possesses a 1 020 000 bp genome with 975 ORF and is deficient in the production of threonine, isoleucine and ubiquinone. Prochlorococcus marinus possesses a 1 751 080 bp genome with 1884 ORF and has a complete metabolism with no deficiencies. The current minimal cell model requires a genome to be of limited size, culturalble and having minimal media requirements as such it is the conclusion of this review that P. marinus best fits this model. Further, future research should concentrate on genome reduction experiments using P. marinus and the search for additional minimal genomes should concentrate on autotrophic free-living organisms. 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The microbial production increased 1.6-fold (thymidine uptake) and 2.1-fold (leucine uptake), while total cell numbers increased only slightly over the course of the experiment. 454 tag pyrosequencing of partial 16S rRNA genes and catalyzed reporter deposition fluorescence in situ hybridization (CARD FISH) showed that the composition and abundance of the bacterial and archaeal community in the iron-fertilized water body were remarkably constant without development of typical bloom-related succession patterns. Members of groups usually found in phytoplankton blooms, such as Roseobacter and Gammaproteobacteria, showed no response or only a minor response to the bloom. However, sequence numbers and total cell numbers of the SAR11 and SAR86 clades increased slightly but significantly toward the end of the experiment. 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Moran, Mary Ann TI Evolution of Divergent Life History Strategies in Marine Alphaproteobacteria SO MBIO DT Article AB Marine bacteria in the Roseobacter and SAR11 lineages successfully exploit the ocean habitat, together accounting for similar to 40% of bacteria in surface waters, yet have divergent life histories that exemplify patch-adapted versus free-living ecological roles. Here, we use a phylogenetic birth-and-death model to understand how genome content supporting different life history strategies evolved in these related alphaproteobacterial taxa, showing that the streamlined genomes of free-living SAR11 were gradually downsized from a common ancestral genome only slightly larger than the extant members (similar to 2,000 genes), while the larger and variably sized genomes of roseobacters evolved along dynamic pathways from a sizeable common ancestor (similar to 8,000 genes). Genome changes in the SAR11 lineage occurred gradually over similar to 800 million years, whereas Roseobacter genomes underwent more substantial modifications, including major periods of expansion, over similar to 260 million years. The timing of the first Roseobacter genome expansion was coincident with the predicted radiation of modern marine eukaryotic phytoplankton of sufficient size to create nutrient-enriched microzones and is consistent with present-day ecological associations between these microbial groups. We suggest that diversification of red-lineage phytoplankton is an important driver of divergent life history strategies among the heterotrophic bacterioplankton taxa that dominate the present-day ocean. IMPORTANCE One-half of global primary production occurs in the oceans, and more than half of this is processed by heterotrophic bacterioplankton through the marine microbial food web. The diversity of life history strategies that characterize different bacterioplankton taxa is an important subject, since the locations and mechanisms whereby bacteria interact with seawater organic matter has effects on microbial growth rates, metabolic pathways, and growth efficiencies, and these in turn affect rates of carbon mineralization to the atmosphere and sequestration into the deep sea. Understanding the evolutionary origins of the ecological strategies that underlie biochemical interactions of bacteria with the ocean system, and which scale up to affect globally important biogeochemical processes, will improve understanding of how microbial diversity is maintained and enable useful predictions about microbial response in the future ocean. C1 [Luo, Haiwei; Moran, Mary Ann] Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. [Cusros, Miklos] Univ Montreal, Dept Comp Sci & Operat Res, Montreal, PQ, Canada. 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Tripp, H. James Givan, Scott A. Smith, Daniel P. Giovannoni, Stephen J. TI Natural variation in SARII marine bacterioplankton genomes inferred from metagenomic data SO BIOLOGY DIRECT DT Article AB Background: One objective of metagenomics is to reconstruct information about specific uncultured organisms from fragmentary environmental DNA sequences. We used the genome of an isolate of the marine alphaproteobacterium SARII ('Candidatus Pelagibacter ubique'; strain HTCC1062), obtained from the cold, productive Oregon coast, as a query sequence to study variation in SARII metagenome sequence data from the Sargasso Sea, a warm, oligotrophic ocean gyre. Results: The average amino acid identity of SARII genes encoded by the metagenomic data to the query genome was only 71%, indicating significant evolutionary divergence between the coastal isolates and Sargasso Sea populations. However, an analysis of gene neighbors indicated that SARII genes in the Sargasso Sea metagenomic data match the gene order of the HTCC1062 genome in 96% of cases (> 85,000 observations), and that rearrangements are most frequent at predicted operon boundaries. There were no conserved examples of genes with known functions being found in the coastal isolates, but not the Sargasso Sea metagenomic data, or vice versa, suggesting that core regions of these diverse SARII genomes are relatively conserved in gene content. However, four hypervariable regions were observed, which may encode properties associated with variation in SARII ecotypes. The largest of these, HVR2, is a 48 kb region flanked by the sole 5S and 23S genes in the HTCC1062 genome, and mainly encodes genes that determine cell surface properties. A comparison of two closely related 'Candidatus Pelagibacter' genomes (HTCC1062 and HTCC1002) revealed a number of "gene indels" in core regions. 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Choi, Chang Jae Worden, Alexandra Z. Baetge, Nicholas Carlson, Craig A. Giovannoni, Stephen TI Seasonality of the Microbial Community Composition in the North Atlantic SO FRONTIERS IN MARINE SCIENCE DT Article AB Planktonic communities constitute the basis of life in marine environments and have profound impacts in geochemical cycles. In the North Atlantic, seasonality drives annual transitions in the ecology of the water column. Phytoplankton bloom annually in spring as a result of these transitions, creating one of the major biological pulses in productivity on earth. The timing and geographical distribution of the spring bloom as well as the resulting biomass accumulation have largely been studied using the global capacity of satellite imaging. However, fine-scale variability in the taxonomic composition, spatial distribution, seasonal shifts, and ecological interactions with heterotrophic bacterioplankton has remained largely uncharacterized. 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Sjostedt, Johanna Casini, Michele Andersson, Agneta Legrand, Catherine Pinhassi, Jarone TI Local Environmental Conditions Shape Generalist But Not Specialist Components of Microbial Metacommunities in the Baltic Sea SO FRONTIERS IN MICROBIOLOGY DT Article AB Marine microbes exhibit biogeographical patterns linked with fluxes of matter and energy. Yet, knowledge of the mechanisms shaping bacterioplankton community assembly across temporal scales remains poor. We examined bacterioplankton 16S rRNA gene fragments obtained from Baltic Sea transects to determine phylogenetic relatedness and assembly processes coupled with niche breadth. Communities were phylogenetically more related over time than expected by chance, albeit with considerable temporal variation. Hence, habitat filtering, i.e., local environmental conditions, rather than competition structured bacterioplankton communities in summer but not in spring or autumn. Species sorting (SS) was the dominant assembly process, but temporal and taxonomical variation in mechanisms was observed. For May communities, Cyanobacteria, Actinobacteria, Alpha- and Betaproteobacteria exhibited SS while Bacteroidetes and Verrucomicrobia were assembled by SS and mass effect. Concomitantly, Gammaproteobacteria were assembled by the neutral model and patch dynamics. Temporal variation in habitat filtering and dispersal highlights the impact of seasonally driven reorganization of microbial communities. Typically abundant Baltic Sea populations such as the NS3a marine group (Bacteroidetes) and the SAR86 and SAR11 clade had the highest niche breadth. The verrucomicrobial Spartobacteria population also exhibited high niche breadth. Surprisingly, variation in bacterioplankton community composition was regulated by environmental factors for generalist taxa but not specialists. 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The microbial communities in the environment are immensely diverse and critical mediators of various ecosystem processes. But it is not clearly known to what extent xenobiotics effect the composition and functionality of environmental microorganisms, across biomes in different geographic locations, with varying contaminating sources, categories of chemical pollutants, and their application types. Here, we investigated the impact of xenobiotics by meta-genomic analysis of 16S rRNA from seawater, soil, sludge and sediment, for deciphering microbial diversity, community structure, functional enrichment and metabolic potentiality in relation to the application of pesticide (metaldehyde), herbicide (2,4-dichlorophenoxyacetic acid; DCPAA), insecticide (chlordecone) and flame retar-dant additives (organophosphate triesters; OPT and tris 2 chloroethyl phosphate; TCEP). Firmicutes was the most abundant phylum consisting of Clostridia, in association with DCPAA (Lutispora sp.), metaldehyde and TCEP treatments. The next two abundant phyla were Proteobacteria [class Deltaproteobacteria, Alphaproteobacteria (Pelagibacter ubique with OPT treatment) and Gammaproteobacteria], and Euryarchaeota consisting mostly of Methanobacteria with chlordecone and DCPAA. Both Desulfomicrobium and Sedimentibacter were predominant in the subtropical moist forest sediment, while Synechococcus and Pelagibacter dominated the temperate Mediterranean Sea. Ruminococcus and Lutispora were more abundant in rain forest compared to waste water solid, while Anaerovorax predominated subtropical moist forest. Sequences involved in house-keeping genes comprised the most abundant hits. The genes involved in environmental information processing such as ABC transporters catalyzing carbohydrate uptake, membrane transport and quorum sensing were over-represented with TCEP, DCPAA, OPT and metaldehyde. Exposure to different classes of xenobiotics revealed major differ-ences in microbial structure and function abundances, mainly driven by environmental sources and biomes. Preponderance of core metabolic functional genes across all the metagenomes analysed is indicative of microbial community responding and adjusting itself to deal with these treatments and maintain its balance. C1 [Mandal, Manisha] MGM Med Coll, Dept Physiol, Kishanganj 855107, India. [Mandal, Shyamapada] Univ Gour Banga, Dept Zool, Malda 732103, India. RP Mandal, S (corresponding author), Univ Gour Banga, Dept Zool, Malda 732103, India. 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Author summary Marine bacteria utilize carbon as a building block and an energy source and thus exert an important control on the amount of carbon that is sequestered in the ocean versus respired into the atmosphere. They use a spectrum of strategies to consume carbon: while copiotrophic bacteria dominate in nutrient-rich environments, oligotrophic bacteria dominate in nutrient-poor environments and are typically smaller, nonmotile, and slower growing. Yet the paragon oligotroph SAR11 is the planet's most abundant organism. Despite this, most of our understanding of bacteria derives from research on copiotrophs. Here we use molecular-level models to understand how an oligotroph's physiology enables it to outperform copiotrophs in nutrient-poor but not in nutrient-rich environments. We contrast copiotrophs' prevalent method of sugar transport with oligotrophs' reliance on binding proteins, which trap nutrients in the periplasm. Binding proteins allow cells to attain affinities that are much higher than the transport proteins' intrinsic affinities. However, our model predicts that attaining such high affinities requires large periplasms with high abundances of the slowly diffusing binding proteins, which precludes high growth rates. By quantifying the benefits and costs of binding proteins, we provide a mechanistic explanation for the divergent evolution of oligotrophs and copiotrophs. C1 [Norris, Noele] MIT, Dept Elect Engn & Comp Sci, Cambridge, MA 02139 USA. [Norris, Noele; Levine, Naomi M.] Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90007 USA. [Norris, Noele; Fernandez, Vicente I.; Stocker, Roman] Swiss Fed Inst Technol, Dept Civil Environm & Geomat Engn, Inst Environm Engn, Zurich, Switzerland. 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Viruses that infect the globally abundant SAR11 bacteria (pelagiphages) were reported to be an important component of the marine viral communities. Our current knowledge of pelagiphages is based on a few studies and therefore is limited. In this study, 10 new pelagiphages were isolated and genomically characterized. These pelagiphages represent the first cultivated representatives of four viral lineages only found in metagenomic sequencing datasets previously. Many abundant environmental viral sequences, i.e., single-virus vSAG 37-F6 and several Global Ocean Viromes (GOV) viral populations, are now further confirmed with these pelagiphages. Viromic read mapping reveals that these new pelagiphages are globally distributed in the ocean and can be detected throughout the water column. Remarkably, isolation of these pelagiphages contributed up to 12% of all viromic reads annotated in the analysed viromes. 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To date the origin of such recalcitrant DOC (RDOC) is unclear. A recently proposed conceptual framework, the microbial carbon pump (MCP), emphasizes the microbial transformation of organic carbon from labile to recalcitrant states. The MCP is concerned with both microbial uptakes and outputs of DOC compounds, covering a wide range from gene to ecosystem levels. In this minireview, the ATP binding cassette (ABC) transporter is used as an example for the microbial processing of DOC at the genetic level. The compositions of the ABC transporter genes of the two major marine bacterial clades Roseobacter and SAR11 demonstrate that they have distinct patterns in DOC utilization: Roseobacter strains have the advantage of taking up carbohydrate DOC, while SAR11 bacteria prefer nitrogen-containing DOC. At the ecosystem level, bacterially derived RDOC based on D-amino acid biomarkers is reported to be responsible for about a quarter of the total marine RDOC pool. 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Previous studies have demonstrated that these riboswitches have the potential for digital modulation of gene expression at both the transcriptional and translational levels. In this study, we investigate the conformational changes in the tandem SAM-II/SAM-V riboswitch binding to S-adenosylmethionine (SAM) using selective 2 '-hydroxyl acylation analyzed by the primer extension (SHAPE) assay, small-angle X-ray scattering (SAXS), and oligos depressing probing. Our findings reveal that the linker between SAM-II/SAM-V aptamers blocks the SAM response of the SAM-II domain. This result proposes a new mechanism for gene expression regulation, where the ligand-binding functions of tandem riboswitches can be selectively masked or released through a linker. C1 [Feng, Shanshan; Xiao, Wenwen; Yu, Yingying; Zhang, Yunlong; Chen, Ting; Lu, Changrui] Donghua Univ, Coll Biol Sci & Med Engn, Shanghai 201620, Peoples R China. 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Crombie, Andrew Scrivens, James H. Murrell, J. Colin TI Bacterial flavin-containing monooxygenase is trimethylamine monooxygenase SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Flavin-containing monooxygenases (FMOs) are one of the most important monooxygenase systems in Eukaryotes and have many important physiological functions. FMOs have also been found in bacteria; however, their physiological function is not known. Here, we report the identification and characterization of trimethylamine (TMA) monooxygenase, termed Tmm, from Methylocella silvestris, using a combination of proteomic, biochemical, and genetic approaches. This bacterial FMO contains the FMO sequence motif (FXGXXXHXXXF/Y) and typical flavin adenine dinucleotide and nicotinamide adenine dinucleotide phosphate-binding domains. The enzyme was highly expressed in TMA-grown M. silvestris and absent during growth on methanol. The gene, tmm, was expressed in Escherichia coli, and the purified recombinant protein had high Tmm activity. Mutagenesis of this gene abolished the ability of M. silvestris to grow on TMA as a sole carbon and energy source. Close homologs of tmm occur in many Alphaproteobacteria, in particular Rhodobacteraceae (marine Roseobacter clade, MRC) and the marine SAR11 clade (Pelagi-bacter ubique). We show that the ability of MRC to use TMA as a sole carbon and/or nitrogen source is directly linked to the presence of tmm in the genomes, and purified Tmm of MRC and SAR11 from recombinant E. coli showed Tmm activities. The tmm gene is highly abundant in the metagenomes of the Global Ocean Sampling expedition, and we estimate that 20% of the bacteria in the surface ocean contain tmm. Taken together, our results suggest that Tmm, a bacterial FMO, plays an important yet overlooked role in the global carbon and nitrogen cycles. 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Najdek, Mirjana TI Temporal variation in the prokaryotic community of a nearshore marine environment SO SCIENTIFIC REPORTS DT Article AB Prokaryotic communities inhabiting surface waters of temperate areas exhibit patterns of seasonal succession. Generally, studies describing these temporal changes are not performed in the proximity to the coast. In the present study, temporal variation of these communities was determined in surface waters at two stations located in the close proximity to the eastern shore of the northern Adriatic Sea. Sequencing of the V4 region of the 16S rRNA gene identified the highest community richness in December with distinct shifts in community structure between periods from April to May, June to October, and November to March. Temperature was shown to be the main environmental force explaining community temporal variation. The NS5 marine group, uncultured Cryomorphaceae, SAR86 clade, and Synechococcus were present throughout the year. Members without know relatives within Rhodobacteraceae and the NS4 marine group were more pronounced in the period from April to May, the AEGEAN-169 marine group, SAR11 subclade III, and HIMB11 in the period from June to October, and SAR11 subclade Ia and Archaea in the period from November to March. Litoricola and OM60 (NOR5) clade were characteristic for both the community sampled from April to May and November to March. Taken together, prokaryotic communities inhabiting nearshore surface waters exhibit a general pattern in community structure similar to other surface associated assemblages of temperate areas. However, the identified specific community composition and temporal patterns differ from other coastal areas. C1 [Korlevic, Marino; Markovski, Marsej; Najdek, Mirjana] Rudjer Boskovic Inst, Ctr Marine Res, Rovinj, Croatia. [Herndl, Gerhard J.] Univ Vienna, Dept Funct & Evolutionary Ecol, Vienna, Austria. 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Colin Chen, Yin TI Trimethylamine N-oxide metabolism by abundant marine heterotrophic bacteria SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Trimethylamine N-oxide (TMAO) is a common osmolyte found in a variety of marine biota and has been detected at nanomolar concentrations in oceanic surface waters. TMAO can serve as an important nutrient for ecologically important marine heterotrophic bacteria, particularly the SAR11 clade and marine Roseobacter clade (MRC). However, the enzymes responsible for TMAO catabolism and the membrane transporter required for TMAO uptake into microbial cells have yet to be identified. We show here that the enzyme TMAO demethylase (Tdm) catalyzes the first step in TMAO degradation. This enzyme represents a large group of proteins with an uncharacterized domain (DUF1989). The function of TMAO demethylase in a representative from the SAR11 clade (strain HIMB59) and in a representative of the MRC (Ruegeria pomeroyi DSS-3) was confirmed by heterologous expression of tdm (the gene encoding Tdm) in Escherichia coli. In R. pomeroyi, mutagenesis experiments confirmed that tdm is essential for growth on TMAO. We also identified a unique ATP-binding cassette transporter (TmoXWV) found in a variety of marine bacteria and experimentally confirmed its specificity for TMAO through marker exchange mutagenesis and lacZ reporter assays of the promoter for genes encoding this transporter. Both Tdm and TmoXWV are particularly abundant in natural seawater assemblages and actively expressed, as indicated by a number of recent metatranscriptomic and metaproteomic studies. These data suggest that TMAO represents a significant, yet overlooked, nutrient for marine bacteria. C1 [Lidbury, Ian; Chen, Yin] Univ Warwick, Sch Life Sci, Coventry CV4 7AL, W Midlands, England. 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To do this, we analyzed population fluctuations and growth responses of major phylogenetic groups of free-living marine bacteria. We used bromodeoxyuridine immunocytochemistry-fluorescence in situ hybridization methodology to examine marine bacterial community development during and after a diatom bloom in a mesocosm. We revealed that the Roseobacter/Rhodobacter, SAR11, Alteromonas, and Bacteroidetes groups were clearly major phylotypes responsible for most free-living bacterial biomass and production throughout the experiment. The clearest bacterial response was a proliferation of the Alteromonas group (cells with large volumes) during development of the bloom (up to 30 % of actively growing cells). Populations of these bacteria declined sharply thereafter, likely due to grazing. Alteromonas group responses suggest that these bacteria strongly influenced the flux of organic matter at an early bloom stage. The growth potential of Bacteroidetes was relatively large as the bloom peaked; this early development probably contributed to the initial stage of bloom decomposition. In contrast, the contribution of Roseobacter/Rhodobacter to total bacterial production increased at a late stage of decomposing of the bloom. The contributions of Betaproteobacteria, SAR11, and SAR86 groups to total bacterial abundance and production were relatively minor throughout the experiment. These results imply that the ability to utilize organic matter derived from diatoms varies among bacterial phylotypes, and, frequently, less abundant but ecological specialist taxa such as Alteromonas may play major roles in the flux of organic matter during diatom blooms. C1 [Tada, Yuya; Hamasaki, Koji] Univ Tokyo, Atmosphere & Ocean Res Inst, Kashiwa, Chiba 2778564, Japan. [Taniguchi, Akito] Kinki Univ, Grad Sch Agr, Naka 6318505, Japan. 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Because of a low pKa value of the retinal Schiff base (RSB), TAT rhodopsin exhibits both a visible light -absorbing state with the protonated RSB and a UV -absorbing state with the deprotonated RSB at a neutral pH. The UV -absorbing state, in contrast to the visible light -absorbing one, converts to a long-lived photointermediate upon light absorption, implying that TAT rhodopsin functions as a pH -dependent light sensor. Despite detailed biophysical characterization and mechanistic studies on the TAT rhodopsin, it has been unknown whether other proteins with similarly unusual features exist. Here, we identified several new rhodopsin genes homologous to the TAT rhodopsin of HIMB114 (TATHIMB) from metagenomic data. Based on the absorption spectra of expressed proteins from these genes with visible and UV peaks similar to that of TATHIMB, they were classified as Twin -peaked Rhodopsin (TwR) family. 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TI Comparison between MICRO-CARD-FISH and 16S rRNA gene clone libraries to assess the active versus total bacterial community in the coastal Arctic SO ENVIRONMENTAL MICROBIOLOGY REPORTS DT Article AB We collected surface- and deep-water samples (maximum depth 300m) during the springsummer transition in the coastal Arctic along a transect in the Kongsfjorden (Ny-angstrom lesund, Spitsbergen, Norway) to determine the structure of the active versus total marine bacterioplankton community using different approaches. Catalysed reporter depositionfluorescence in situ hybridization combined with microautoradiography (MICROCARDFISH) was used to determine the abundance and activity of different bacterial groups. The bacterial communities were dominated by members of Alphaproteobacteria followed by Bacteroidetes, whereas Gammaproteobacteria were present at low abundance but exhibited a high percentage of active cells taking up leucine. The clone libraries of 16S rRNA genes (16S rDNA) and 16S rRNA from two different depths were used to decipher the bacterial community structure. Independently of the type of clone libraries analysed (16S rDNA- or 16S rRNA-based), four major and four minor taxonomic groups were detected. The bacterioplankton community was mainly dominated at both the DNA and the RNA levels by Alphaproteobacteria followed by Gammaproteobacteria. The Rhodobacteriaceae were the most abundant members of the Alphaproteobacteria in both DNA and RNA clone libraries, followed by the SAR11 clade, which was only detectable at the 16S rDNA level. Moreover, there was a general agreement between the results obtained with both techniques, although some specific phylogenetic groups, such as SAR11 and Roseobacter, deviated substantially from this relation. These discrepancies are most likely linked to different physiological states among members of the bacterioplankton community. 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Article AB Fjords are sensitive areas affected by climate change and can act as a natural laboratory to study microbial ecological processes. The Chilean Patagonian fjords (41-56 degrees S), belonging to the Subantarctic ecosystem (46-60 degrees S), make up one of the world's largest fjord systems. In this region, Estuarine Water (EW) strongly influences oceanographic conditions, generating sharp gradients of oxygen, salinity and nutrients, the effects of which on the microbial community structure are poorly understood. During the spring of 2017 we studied the ecological patterns (dispersal and oceanographic factors) underlying the microbial community distribution in a linear span of 450 km along the estuarine-influenced Chilean Patagonian fjords. Our results show that widespread microbial dispersion existed along the fjords where bacterioplankton exhibited dependence on the eukaryotic phytoplankton community composition. This dependence was particularly observed under the low chlorophyll-a conditions of the Baker Channel area, in which a significant relationship was revealed between SAR11 Clade III and the eukaryotic families Pyrenomonadaceae (Cryptophyte) and Coccomyxaceae (Chlorophyta). Furthermore, dissolved oxygen and salinity were revealed as the main drivers influencing the surface marine microbial communities in these fjords. A strong salinity gradient resulted in the segregation of the Baker Channel prokaryotic communities from the rest of the Patagonian fjords. Likewise, Microbacteriaceae, Burkholderiaceae and SAR11 Clade III, commonly found in freshwater, were strongly associated with EW conditions in these fjords. The direct effect of EW on the microbial community structure and diversity of the fjords exemplifies the significance that climate change and, in particular, deglaciation have on this marine region and its productivity. C1 [Tamayo-Leiva, Javier; Cifuentes-Anticevic, Jeronimo; Arroyo, Jose Ignacio; Diez, Beatriz] Pontificia Univ Catolica Chile, Fac Biol Sci, Dept Mol Genet & Microbiol, Santiago, Chile. [Tamayo-Leiva, Javier; Aparicio-Rizzo, Pilar; Masotti, Italo; Diez, Beatriz] Univ Chile, Ctr Climate & Resilience Res CR2, Santiago, Chile. [Aparicio-Rizzo, Pilar; Masotti, Italo] Univ Valparaiso, Fac Ciencias Mar & Recursos Nat, Vina Del Mar, Chile. [Arroyo, Jose Ignacio] Santa Fe Inst, Santa Fe, NM 87501 USA. [Arroyo, Jose Ignacio] Pontificia Univ Catolica Chile, Fac Biol Sci, Dept Ecol, Santiago, Chile. [Arroyo, Jose Ignacio] Univ Chile, Ctr Math Modeling, Santiago, Chile. [Masotti, Italo] Univ Valparaiso, Ctr Observ Marino Estudios Riesgos Ambiente Coste, Vina Del Mar, Chile. [Diez, Beatriz] Ctr Genome Regulat CRG, Santiago, Chile. 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Based on terminal restriction fragment length polymorphism analysis of small-subunit rRNA genes, similarities in bacterial communities associated with 170-h-old planulae were observed regardless of treatment, suggesting that bacteria may have been externally associated from the outset of the experiment. Microscopic examination of P. meandrina planulae by fluorescence in situ hybridization with bacterial and Roseobacter clade-specific oligonucleotide probes revealed differences in the densities and locations of planulae-associated cells. Planulae exposed to either raw seawater or strains of Pseudoalteromonas and Roseobacter harbored the highest densities of internally associated cells, of which 20 to 100% belonged to the Roseobacter clade. Planulae exposed to sterile seawater or strains of the SAR11 clade and Synechococcus did not show evidence of prominent bacterial associations. 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Eiler, Alexander Herfort, Lydie Needoba, Joseph A. Peterson, Tawnya D. Crump, Byron C. TI Determining indicator taxa across spatial and seasonal gradients in the Columbia River coastal margin SO ISME JOURNAL DT Article AB Bacterioplankton communities are deeply diverse and highly variable across space and time, but several recent studies demonstrate repeatable and predictable patterns in this diversity. We expanded on previous studies by determining patterns of variability in both individual taxa and bacterial communities across coastal environmental gradients. We surveyed bacterioplankton diversity across the Columbia River coastal margin, USA, using amplicon pyrosequencing of 16S rRNA genes from 596 water samples collected from 2007 to 2010. Our results showed seasonal shifts and annual reassembly of bacterioplankton communities in the freshwater-influenced Columbia River, estuary, and plume, and identified indicator taxa, including species from freshwater SAR11, Oceanospirillales, and Flavobacteria groups, that characterize the changing seasonal conditions in these environments. In the river and estuary, Actinobacteria and Betaproteobacteria indicator taxa correlated strongly with seasonal fluctuations in particulate organic carbon (rho = -0.664) and residence time (rho=0.512), respectively. In contrast, seasonal change in communities was not detected in the coastal ocean and varied more with the spatial variability of environmental factors including temperature and dissolved oxygen. Indicator taxa of coastal ocean environments included SAR406 and SUP05 taxa from the deep ocean, and Prochlorococcus and SAR11 taxa from the upper water column. We found that in the Columbia River coastal margin, freshwater-influenced environments were consistent and predictable, whereas coastal ocean community variability was difficult to interpret due to complex physical conditions. This study moves beyond beta-diversity patterns to focus on the occurrence of specific taxa and lends insight into the potential ecological roles these taxa have in coastal ocean environments. C1 [Fortunato, Caroline S.; Crump, Byron C.] Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD USA. [Eiler, Alexander] Uppsala Univ, Dept Ecol & Genet, Uppsala, Sweden. [Herfort, Lydie; Needoba, Joseph A.; Peterson, Tawnya D.] Oregon Hlth & Sci Univ, Ctr Coastal Margin Observat & Predict, Beaverton, OR USA. RP Fortunato, CS (corresponding author), Josephine Bay Paul Ctr, Marine Biol Lab, Woods Hole, MA 02543 USA. 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TI Contrasting diversity patterns of prokaryotes and protists over time and depth at the San-Pedro Ocean Time series SO ISME COMMUNICATIONS DT Article AB Community dynamics are central in microbial ecology, yet we lack studies comparing diversity patterns among marine protists and prokaryotes over depth and multiple years. Here, we characterized microbes at the San-Pedro Ocean Time series (2005-2018), using SSU rRNA gene sequencing from two size fractions (0.2-1 and 1-80 mu m), with a universal primer set that amplifies from both prokaryotes and eukaryotes, allowing direct comparisons of diversity patterns in a single set of analyses. The 16S + 18S rRNA gene composition in the small size fraction was mostly prokaryotic (>92%) as expected, but the large size fraction unexpectedly contained 46-93% prokaryotic 16S rRNA genes. Prokaryotes and protists showed opposite vertical diversity patterns; prokaryotic diversity peaked at mid-depth, protistan diversity at the surface. 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The first enzyme of this pathway, glyoxalase I (GlxI), uses methylglyoxal as a substrate and requires either Ni(II)/Co(II) or Zn(II) for activity. Here we have investigated the origin of different metal ion specificities of GlxI and subsequent pattern of inheritance during evolution. Our results suggest a primitive origin of single-domain Ni dependent GlxI [Ni-GlxI]. This subsequently evolved into Zn activated GlxI [Zn-GlxI] in deltaproteobacteria. However, origin of eukaryotic Zn-GlxI is different and can be traced to GlxI from Candidatus pelagibacter and Sphingomonas. In eukaryotes GlxI has evolved as two-domain protein but the corresponding Zn form is lost in plants/higher eukaryotes. In plants gene expansion has given rise to multiple two-domain Ni-GlxI which are differentially regulated under abiotic stress conditions. Our results suggest that different forms of GlxI have evolved to help plants adapt to stress. 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TI Adaptive genetic traits in pelagic freshwater microbes SO ENVIRONMENTAL MICROBIOLOGY DT Review AB Pelagic microbes have adopted distinct strategies to inhabit the pelagial of lakes and oceans and can be broadly categorized in two groups: free-living, specialized oligotrophs and patch-associated generalists or copiotrophs. In this review, we aim to identify genomic traits that enable pelagic freshwater microbes to thrive in their habitat. To do so, we discuss the main genetic differences of pelagic marine and freshwater microbes that are both dominated by specialized oligotrophs and the difference to freshwater sediment microbes, where copiotrophs are more prevalent. We phylogenomically analysed a collection of >7700 metagenome-assembled genomes, classified habitat preferences on different taxonomic levels, and compared the metabolic traits of pelagic freshwater, marine, and freshwater sediment microbes. 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Sebastian, Marta Hojerova, Eva Koblizek, Michal TI Comparison of Growth Rates of Aerobic Anoxygenic Phototrophic Bacteria and Other Bacterioplankton Groups in Coastal Mediterranean Waters SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Growth is one of the basic attributes of any living organism. Surprisingly, the growth rates of marine bacterioplankton are only poorly known. Current data suggest that marine bacteria grow relatively slowly, having generation times of several days. However, some bacterial groups, such as the aerobic anoxygenic phototrophic (AAP) bacteria, have been shown to grow much faster. Two manipulation experiments, in which grazing, viruses, and resource competition were reduced, were conducted in the coastal Mediterranean Sea (Blanes Bay Microbial Observatory). The growth rates of AAP bacteria and of several important phylogenetic groups (the Bacteroidetes, the alphaproteobacterial groups Roseobacter and SAR11, and the Gammaproteobacteria group and its subgroups the Alteromonadaceae and the NOR5/OM60 clade) were calculated from changes in cell numbers in the manipulation treatments. In addition, we examined the role that top-down (mortality due to grazers and viruses) and bottom-up (resource availability) factors play in determining the growth rates of these groups. Manipulations resulted in an increase of the growth rates of all groups studied, but its extent differed largely among the individual treatments and among the different groups. Interestingly, higher growth rates were found for the AAP bacteria (up to 3.71 day(-1)) and for the Alteromonadaceae (up to 5.44 day(-1)), in spite of the fact that these bacterial groups represented only a very low percentage of the total prokaryotic community. 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Coe, Allison Arellano, Aldo A. Dooley, Keven Silvestri, Samantha M. Gong, Jacqueline S. Yeager, Emily A. Becker, Jamie W. Chisholm, Sallie W. TI Environmental and Taxonomic Drivers of Bacterial Extracellular Vesicle Production in Marine Ecosystems SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Bacteria release extracellular vesicles that contain a wide variety of cellular compounds, including lipids, proteins, nucleic acids, and small molecules, into their surrounding environment. These structures are found in diverse microbial habitats, including the oceans, where their distributions vary throughout the water column and likely affect their functional impacts within microbial ecosystems. Extracellular vesicles are small (approximately 50 to 250 nm in diameter), membrane-bound structures that are released by cells into their surrounding environment. Heterogeneous populations of vesicles are abundant in the global oceans, and they likely play a number of ecological roles in these microbially dominated ecosystems. Here, we examine how vesicle production and size vary among different strains of cultivated marine microbes as well as explore the degree to which this is influenced by key environmental variables. We show that both vesicle production rates and vesicle sizes significantly differ among cultures of marine Proteobacteria, Cyanobacteria, and Bacteroidetes. Further, these properties vary within individual strains as a function of differences in environmental conditions, such as nutrients, temperature, and light irradiance. Thus, both community composition and the local abiotic environment are expected to modulate the production and standing stock of vesicles in the oceans. Examining samples from the oligotrophic North Pacific Gyre, we show depth-dependent changes in the abundance of vesicle-like particles in the upper water column in a manner that is broadly consistent with culture observations: the highest vesicle abundances are found near the surface, where the light irradiances and the temperatures are the greatest, and they then decrease with depth. This work represents the beginnings of a quantitative framework for describing extracellular vesicle dynamics in the oceans, which is essential as we begin to incorporate vesicles into our ecological and biogeochemical understanding of marine ecosystems.IMPORTANCE Bacteria release extracellular vesicles that contain a wide variety of cellular compounds, including lipids, proteins, nucleic acids, and small molecules, into their surrounding environment. These structures are found in diverse microbial habitats, including the oceans, where their distributions vary throughout the water column and likely affect their functional impacts within microbial ecosystems. Using a quantitative analysis of marine microbial cultures, we show that bacterial vesicle production in the oceans is shaped by a combination of biotic and abiotic factors. Different marine taxa release vesicles at rates that vary across an order of magnitude, and vesicle production changes dynamically as a function of environmental conditions. These findings represent a step forward in our understanding of bacterial extracellular vesicle production dynamics and provide a basis for the quantitative exploration of the factors that shape vesicle dynamics in natural ecosystems. C1 [Biller, Steven J.; Coe, Allison; Arellano, Aldo A.; Dooley, Keven; Becker, Jamie W.; Chisholm, Sallie W.] MIT, Dept Civil & Environm Engn, Cambridge, MA 02139 USA. 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Potvin, Marianne Casamayor, Emilio O. Lovejoy, Connie TI Hydrography shapes bacterial biogeography of the deep Arctic Ocean SO ISME JOURNAL DT Article AB It has been long debated as to whether marine microorganisms have a ubiquitous distribution or patterns of biogeography, but recently a consensus for the existence of microbial biogeography is emerging. However, the factors controlling the distribution of marine bacteria remain poorly understood. In this study, we combine pyrosequencing and traditional Sanger sequencing of the 16S rRNA gene to describe in detail bacterial communities from the deep Arctic Ocean. We targeted three separate water masses, from three oceanic basins and show that bacteria in the Arctic Ocean have a biogeography. The biogeographical distribution of bacteria was explained by the hydrography of the Arctic Ocean and subsequent circulation of its water masses. 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However, its impact on heterotrophic prokaryotic production (HPP) remains unclear due to the complex interplay of multiple indirect factors governing this process. Here, we conducted a comprehensive investigation of the distribution of picophytoplankton and heterotrophic prokaryotes, prokaryotic community structure, and HPP within a cyclonic eddy in the western North Pacific subtropical gyre. The results indicated that despite the higher abundance of picophytoplankton accompanied by nutrient upwelling at the eddy core compared to the edge, higher levels of HPP were observed at the eddy edge between 100 and 200 m, consistent with the distribution of the low nucleic acid content (LNA) prokaryotes. The significant positive correlation between HPP and the proportion of LNA group in total heterotrophic prokaryotes suggested a primary contribution from the LNA group over the high nucleic acid content (HNA) group. SAR11, a typical member of the LNA group, may primarily contribute to the elevated HPP observed at the eddy edge. The changes in temperature, nutrients, and light intensity induced by the cyclonic eddy may significantly influence the distribution and activity of HNA and LNA groups, potentially exerting a greater impact on HPP compared to phytoplankton-related factors. These findings contribute to understanding the underlying mechanisms of HPP responses to cyclonic eddies in the oligotrophic open ocean. C1 [Ni, Junyi; Chen, Mingming; Shen, Jiaming; Liu, Li; Zhou, Kuanbo; Li, Xiaolin; Dai, Minhan; Zhang, Yao] Xiamen Univ, Coll Ocean & Earth Sci, State Key Lab Marine Environm Sci, Xiamen, Peoples R China. [Jiang, Zong-Pei] Zhejiang Univ, Ocean Coll, Zhoushan, Peoples R China. RP Zhang, Y (corresponding author), Xiamen Univ, Coll Ocean & Earth Sci, State Key Lab Marine Environm Sci, Xiamen, Peoples R China. 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Pellizari, Vivian H. Enrich-Prast, Alex Sievert, Stefan M. TI Spatiotemporal dynamics of marine bacterial and archaeal communities in surface waters off the northern Antarctic Peninsula SO DEEP-SEA RESEARCH PART II-TOPICAL STUDIES IN OCEANOGRAPHY DT Article AB Seasonal changes in taxonomic and functional diversity of microbial communities in polar regions are commonly observed, requiring strategies of microbes to adapt to the corresponding changes in environmental conditions. These natural fluctuations form the backdrop for changes induced by anthropogenic impacts. The main goal of this study was to assess the seasonal and temporal changes in bacterial and archaeal diversity and community structure off the northern Antarctic Peninsula over several seasons (spring, summer, autumn) from 2013 to 2015. Ten monitoring stations were selected across the Gerlache and Bransfield Straits and nearby Elephant Island, and archaeal and bacterial communities examined by amplicon sequencing of 16S rRNA genes. Alpha-diversity indices were higher in spring and correlated significantly with temperature. Spring was characterized by the presence of SAR11, and microbial communities remaining from winter, including representatives of Thaumarchaeota (Nimosopurnilus), Euryarchaeota, members of Oceanospirillales, SAR324. Summer and autumn were characterized by a high prevalence of Flavobacteria (NS5 marine group and Polaribacter), Alphaproizobacteria (Rhodobacterales and SAR11 Glade) and Gammaproteobacteria (Oceanospirillales/Balneatrix and Celivibrionales), generally known to be associated with organic matter degradation. Relatively higher abundance of phytoplankton groups occurred in spring, mainly characterized by the presence of the haptophyte Phaeocystis and the diatom Corethron, influencing the succession of heterotrophic bacterial communities. Microbial diversity and community structure varied significantly over time, but not over space, i.e., were similar between monitoring stations for the same time. In addition, the observed interannual variability in microbial community structure might be related to an increase in sea surface temperature. Environmental conditions related to seasonal variation, including temperature and most likely phytoplankton derived organic matter, appear to have triggered the observed shifts in microbial communities in the waters off the northern Antarctic Peninsula. C1 [Signori, Camila N.; Pellizari, Vivian H.] Univ Sao Paulo, Inst Oceanog, Dept Oceanog Biol, Praca Oceanog 191, BR-05508900 Sao Paulo, SP, Brazil. 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In order to investigate the significance of distinct phylogenetic bacterial groups it is not only important to assess their quantitative abundance but also their growth dynamics in relation to the entire bacterioplankton. Therefore bacterial abundance, biomass production and the composition of the entire and cell-proliferating bacterioplankton community were assessed in North Sea surface waters between the German Bight and 58 degrees N in early summer by applying catalyzed reporter deposition (CARD-FISH) and bromodeoxyuridine fluorescence in situ hybridization (BrdU-FISH). Bacteroidetes and the Roseobacter group dominated the cell-proliferating fraction with 10-55 and 8-31% of total BrdU-positive cells, respectively. While Bacteroidetes also showed high abundances in the total bacterial fraction, roseobacters constituted only 1-9% of all cells. Despite abundances of up to 55% of total bacterial cells, the SAR11 clade constituted <6% of BrdU-positive cells. Gammaproteobacteria accounted for 2-16% of the total and 2-13% of the cell-proliferating cells. Within the two most active groups, BrdU-positive cells made up 28% of Bacteroidetes as an overall mean and 36% of roseobacters. Estimated mean growth rates of Bacteroidetes and the Roseobacter group were 1.2 and 1.5 day(-1), respectively, and much higher than bulk growth rates of the bacterioplankton whereas those of the SAR11 clade and Gammaproteobacteria were 0.04 and 0.21 day(-1), respectively, and much lower than bulk growth rates. Only numbers of total and cell-proliferating roseobacters but not those of Bacteroidetes and the other groups were significantly correlated to chlorophyll fluorescence and bacterioplankton biomass production. The Roseobacter group, besides Bacteroidetes, appeared to be a major player in processing phytoplankton derived organic matter despite its low partitioning in the total bacterioplankton community. 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The community composition of the sediment did not change over seasons, dominated by Syntrophus species. The insights gained from this study may contribute to understanding how dissolved oxygen, H2S concentrations and salinity drive bacterial community structure in euxinic ecosystems especially the dominance of anoxic bacteria. C1 [Ozbayram, E. Gozde; Koker, Latife; Oguz, Ayca; Zengin, Zuhal; Akcaalan, Reyhan; Albay, Meric] Istanbul Univ, Fac Aquat Sci, Dept Marine & Freshwater Resources Management, TR-34134 Istanbul, Turkiye. RP Ozbayram, EG (corresponding author), Istanbul Univ, Fac Aquat Sci, Dept Marine & Freshwater Resources Management, TR-34134 Istanbul, Turkiye. 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Here, we have performed a cross-platform study of the two most common HTS methodologies, i.e., 454-pyrosequencing and Illumina tags to evaluate their usefulness in biodiversity monitoring and assessment of environmental status. Picoplankton biodiversity has been compared through both methodologies by sequencing the 16 and 18S rRNA genes of a set of samples collected in the coast of Barcelona (NW Mediterranean). The results show that, despite differences observed in the rare OTUs retrieved, both platforms provide a comparable view of the marine picoplankton communities. On a taxonomic level, there was an accurate overlap in the detected phyla between the two methods and the overall estimates of alpha- and betadiversity were comparable. In addition, we explored the concept of "indicator species" and found that certain taxa (i.e., members of the Gammaproteobacteria among others) as well as the ratio between some phylogenetic groups (i.e., the ratio of Alphaproteobacteria/Gammaproteobacteria, Alteromonas/SAR11, and Alteromonas + Oceanospirillales/SAR11) have potential for being useful indicators of environmental status. The data show that implementing new protocols and identifying indicators of environmental status based on rRNA amplicon sequencing is feasible, and that is worth exploring whether the identified indices are universally applicable. C1 [Ferrera, Isabel; Giner, Caterina R.; Rene, Albert; Camp, Jordi; Massana, Ramon; Gasol, Josep M.; Garces, Esther] CSIC, Biol Marina & Oceanog, Inst Ciencies Mar, Barcelona, Spain. RP Ferrera, I (corresponding author), CSIC, Biol Marina & Oceanog, Inst Ciencies Mar, Barcelona, Spain. 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TI Impact of instrument and experiment parameters on reproducibility of ultrahigh resolution ESI FT-ICR mass spectra of natural organic matter SO ORGANIC GEOCHEMISTRY DT Article AB Natural dissolved organic matter (OM) is a complex heterogeneous mixture of compounds that have defied traditional characterization using standard analytical methods. Electrospray ionization mass spectrometry, particularly in ultrahigh resolution mode, provides a new platform for compositional assessment of this important pool of the Earth's reduced carbon. Here, we propose a framework for optimization of instrument and experiment parameters for high quality data acquisition using Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). We report the impact of these parameters on reproducibility of peak detection and repeatability of peak height in replicate injections of Suwannee River fulvic acid (FA), a common terrestrial OM standard. 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Sogin, Mitchell L. Herndl, Gerhard J. TI Water mass-specificity of bacterial communities in the North Atlantic revealed by massively parallel sequencing SO MOLECULAR ECOLOGY DT Article AB Bacterial assemblages from subsurface (100 m depth), meso- (200-1000 m depth) and bathy-pelagic (below 1000 m depth) zones at 10 stations along a North Atlantic Ocean transect from 60 degrees N to 5 degrees S were characterized using massively parallel pyrotag sequencing of the V6 region of the 16S rRNA gene (V6 pyrotags). In a dataset of more than 830 000 pyrotags, we identified 10 780 OTUs of which 52% were singletons. The singletons accounted for less than 2% of the OTU abundance, whereas the 100 and 1000 most abundant OTUs represented 80% and 96% respectively of all recovered OTUs. Non-metric Multi-Dimensional Scaling and Canonical Correspondence Analysis of all the OTUs excluding the singletons revealed a clear clustering of the bacterial communities according to the water masses. 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Bertolino, Paola Vershinin, Konstantin E. Lopez-Garcia, Purificacion TI Marine signature taxa and core microbial community stability along latitudinal and vertical gradients in sediments of the deepest freshwater lake SO ISME JOURNAL DT Article; Early Access AB Lake Baikal is the deepest (similar to 1.6 km) and most voluminous freshwater reservoir on Earth. Compared to plankton, its benthos remains poorly explored. Here, we ask whether latitude and/or depth determine benthic microbial community structure and how Baikal communities compare to those of other freshwater, brackish and marine sediments. To answer, we collected sediment upper layers (0-1 cm) across a similar to 600 km North-South transect covering the three basins of the lake and from littoral to bathybenthic depths (0.5-1450 m). Analysis of 16S and 18S rRNA gene amplicon sequences revealed communities with high richness and evenness where rare operational taxonomic units (OTUs) collectively dominated. Archaea represented up to 25% or prokaryotic sequences. Baikal sediments harbored typically marine eukaryotic and prokaryotic OTUs recently identified in some lakes (diplonemids, Bolidophyceae, Mamiellales, SAR202, marine-like Synechococcus, Pelagibacterales) but also SAR324, Syndiniales and Radiolaria. We hypothesize that, beyond the salinity barrier, adaptation to oligotrophy explains the presence of these otherwise typically marine lineages. Baikal core benthic communities were relatively stable across sites and seemed not determined by depth or latitude. Comparative analyses with other freshwater, brackish and marine prokaryotic sediment communities confirmed the distinctness of Baikal benthos, which include elements of similarity to marine and hydrothermally influenced systems. C1 [Reboul, Guillaume; Moreira, David; Bertolino, Paola; Lopez-Garcia, Purificacion] Univ Paris Saclay, AgroParisTech, Ecol Systemat Evolut, Ctr Natl Rech Sci CNRS, Orsay, France. 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We found selective use of substrates by these groups, although the abundances of Ant4D3 (Antarctic Gammaproteobacteria), Polaribacter (Bacteroidetes), and SAR11 (Alphaproteobacteria) were not different between summer and winter in the Beaufort and Chukchi Seas. The number of cells taking up glucose within all three bacterial groups decreased significantly from summer to winter, while the percentage of cells using leucine did not show a clear pattern between seasons. The uptake of the amino acid mix increased substantially from summer to winter by the Ant4D3 group, although such a large increase in uptake was not seen for the other two groups. Use of glucose by bacteria, but not use of leucine or the amino acid mix, related strongly to inorganic nutrients, chlorophyll a, and other environmental factors. 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It was hypothesised that the composition of the heterotrophic picoplankton would be different in these contrasting waters. To reduce the total diversity, cells were sorted by flow cytometry according to their scatter and DNA content before PCR amplification. The 16S rRNA clone libraries resulting from flow-sorted populations were different and often dominated by a small number of clades. Libraries from the Prochlorococcus-dominated southerly waters were dominated by sequences related to uncultured clusters of SAR11, SAR86 and Actinobacteria (HGC 1). From surface waters of the Synechococcus-dominated northern part of the Arabian Sea, mostly sequences related to the uncultured gammaproteobacterial group 'Svalbard' and HGC I were retrieved. The clone libraries from the OMZ were also dominated by sequences falling in the clades SARI 1 and SAR406, but included sequences related to those of sulfate-reducing (Desulfosarcina, Desulfofrigus) and sulfide-oxidising bacteria (endosymbionts of Riftia and Calyptogena). With a recently developed more sensitive FISH protocol approximately 60% of all DAPI stained cells could be identified by general probes as Bacteria, Cren- or Euryarchaeota in both provinces of the Arabian Sea; 40% remained undetected. On this level and on that of the major phylogenetic groups like Alpha- and Gammaproteobacteria only minor differences were detected by FISH. However, the composition of heterotrophic picoplankton clearly differed for the proteobacterial subgroups SAR86, SAR11 and SAR116. These were more abundant in the oligotrophic waters throughout the water column than in the mesotrophic surface waters and the OMZ. 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Particle-attached bacteria dominate bacterial activity in the estuary and are an important part of the estuarine food web. PCR-amplified 16S rRNA genes from particle-attached and free-living bacteria in the Columbia River, its estuary, and the adjacent coastal ocean were cloned, and 239 partial sequences were determined. A wide diversity was observed at the species level within at least six different bacterial phyla, including most subphyla of the class Proteobacteria. In the estuary, most particle-attached bacterial clones (75%) were related to members of the genus Cytophaga or of the alpha, gamma, or beta subclass of the class Proteobacteria. These same clones, however, were rare in or absent from either the particle-attached or the free-living bacterial communities of the river and the coastal ocean. In contrast, about half (48%) of the free-living estuarine bacterial clones were similar to clones from the river or the coastal ocean. These free-living bacteria were related to groups of cosmopolitan freshwater bacteria (beta-proteobacteria, gram-positive bacteria, and Verrucomicrobium spp,) and groups of marine organisms (gram-positive bacteria and alpha-proteobacteria [SAR11 and Rhodobacter spp.]). These results suggest that rapidly growing particle-attached bacteria develop into a uniquely adapted estuarine community and that free-living estuarine bacteria are similar to members of the river and the coastal ocean microbial communities. The high degree of diversity in the estuary is the result of the mixing of bacterial communities from the river, estuary, and coastal ocean. C1 Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. RP Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. 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Martiny, Adam C. TI The Ocean as a Global Reservoir of Antibiotic Resistance Genes SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Recent studies of natural environments have revealed vast genetic reservoirs of antibiotic resistance (AR) genes. Soil bacteria and human pathogens share AR genes, and AR genes have been discovered in a variety of habitats. However, there is little knowledge about the presence and diversity of AR genes in marine environments and which organisms host AR genes. To address this, we identified the diversity of genes conferring resistance to ampicillin, tetracycline, nitrofurantoin, and sulfadimethoxine in diverse marine environments using functional metagenomics (the cloning and screening of random DNA fragments). Marine environments were host to a diversity of AR-conferring genes. Antibiotic-resistant clones were found at all sites, with 28% of the genes identified as known AR genes (encoding beta-lactamases, bicyclomycin resistance pumps, etc.). However, the majority of AR genes were not previously classified as such but had products similar to proteins such as transport pumps, oxidoreductases, and hydrolases. Furthermore, 44% of the genes conferring antibiotic resistance were found in abundant marine taxa (e.g., Pelagibacter, Prochlorococcus, and Vibrio). Therefore, we uncovered a previously unknown diversity of genes that conferred an AR phenotype among marine environments, which makes the ocean a global reservoir of both clinically relevant and potentially novel AR genes. C1 [Hatosy, Stephen M.; Martiny, Adam C.] Univ Calif Irvine, Dept Ecol & Evolutionary Biol, Irvine, CA 92717 USA. [Martiny, Adam C.] Univ Calif Irvine, Dept Earth Syst Sci, Irvine, CA USA. RP Martiny, AC (corresponding author), Univ Calif Irvine, Dept Ecol & Evolutionary Biol, Irvine, CA 92717 USA. 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Divergent heterotrophic strategies for Alphaproteobacteria and Flavobacteria were indicated by the metaproteome data, with Alphaproteobacteria capturing (by high-affinity transport) and processing labile solutes, and Flavobacteria expressing outer membrane receptors for particle adhesion to facilitate the exploitation of non-labile substrates. TonB-dependent receptors from Gammaproteobacteria and Flavobacteria (particularly in summer) were abundant, indicating that scavenging of substrates was likely an important strategy for these clades of Southern Ocean bacteria. This study provides the first insight into differences in functional processes occurring between summer and winter microbial communities in coastal Antarctic waters, and particularly highlights the important role that 'dark' carbon fixation has in winter. 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When the DOM was more degraded, 16S rRNA gene amplicon data revealed a less diverse bacterioplankton community with a significant contribution from members of the Flavobacteriaceae family. Over the course of 7-10 d, as the DOM quality improved (became less degraded) and bacterioplankton productivity increased, the responding bacterioplankton community became more diverse, with increased relative contributions from members of the SAR86, SAR11 and AEGEAN-169 clades. The cruise mean for mixed layer, depth-integrated bacterioplankton carbon demand (gross bacterioplankton production) was 5.2 mmol C m-2 d-1, representing 60% of net primary production, where the difference between net primary production and bacterioplankton carbon demand was less than sinking flux at 50 m. The concentrations of dissolved organic carbon (cruise average of 58.5 mM C) did not exhibit a systematic change over the cruise period. 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The isolation of phages that infect SAR11 and SAR116 have demonstrated the dominance of these phages in the marine virosphere. However, no phages have been isolated from bacteria in the Roseobacter RCA lineage, another abundant group of marine bacteria. In this study, seven RCA phages that infect three different RCA strains were isolated and characterized. All seven RCA phages belong to the Podoviridae family and have genome sizes ranging from 39.6 to 58.1 kb. Interestingly, three RCA phages (CRP-1, CRP-2, and CRP-3) show similar genomic content and architecture as SAR116 phage HMO-2011, which represents one of the most abundant known viral groups in the ocean. The high degree of homology among CRP-1, CRP-2, CRP-3, and HMO-2011 resulted in the contribution of RCA phages to the dominance of the HMO-2011-type group. CRP-4 and CRP-5 are similar to the Cobavirus group roseophages in terms of gene content and organization. The remaining two RCA phages, CRP-6 and CRP-7, show limited genomic similarity with known phages and represent two new phage groups. Metagenomic fragment recruitment analyses reveal that these RCA phage groups are much more abundant in the ocean than most existing marine roseophage groups. The characterization of these RCA phages has greatly expanded our understanding of the genomic diversity and evolution of marine roseophages and suggests the critical need for isolating phages from the abundant but "unculturable" bacteria. IMPORTANCE The RCA lineage of the marine Roseobacter group represents one of the slow-growing but dominant components of marine microbial communities. Although dozens of roseophages have been characterized, no phages infecting RCA strains have been reported. In this study, we reported on the first RCA phage genomes and investigated their distribution pattern and relative abundance in comparison with other important marine phage groups. Two of the four RCA phage groups were found closely related to previously reported SAR116 phage HMO-2011 and Cobavirus group roseophages, respectively. The remaining two groups are novel in the genome contents. Our study also revealed that RCA phages are widely distributed and exhibit high abundance in marine viromic data sets. Altogether, our findings have greatly broadened our understanding of RCA phages and emphasize the ecological and evolutionary importance of RCA phages in the marine virosphere. C1 [Zhang, Zefeng; Qin, Fang; Zhai, Zhiqiang; Yang, Mingyu; Zhao, Yanlin] Fujian Agr & Forestry Univ, Coll Life Sci, Fujian Prov Key Lab Agroecol Proc & Safety Monito, Fuzhou, Fujian, Peoples R China. [Chen, Feng] Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA. [Chu, Xiao; Zhang, Hao; Luo, Haiwei] Chinese Univ Hong Kong, Sch Life Sci, Simon FS Li Marine Sci Lab, Shatin, Hong Kong, Peoples R China. [Chu, Xiao; Zhang, Hao; Luo, Haiwei] Chinese Univ Hong Kong, State Key Lab Agrobiotechnol, Shatin, Hong Kong, Peoples R China. [Sun, Jing] Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao, Shandong, Peoples R China. RP Zhao, YL (corresponding author), Fujian Agr & Forestry Univ, Coll Life Sci, Fujian Prov Key Lab Agroecol Proc & Safety Monito, Fuzhou, Fujian, Peoples R China. 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Vibrionaceae, Alteromonadaceae, and Flavobacteriaceae dominated in reef-associated sediments, whereas Rickettsiaceae and Synechoccaceae were more highly represented in the water column. Bacterial communities from sediments were more diverse than from the water column. This study reveals cryptic bacterial diversity among microenvironmental components of marine microbial reef communities subject to differential influence of anthropogenic stressors. Such investigations are critical for constructing scenarios of environmentally induced shifts in bacterial biodiversity and species composition. C1 [Rodriguez-Gomez, Citlali; Oliart-Ros, Rosa Maria] Tecnol Nacl Mexico, Inst Tecnol Veracruz, Unidad Invest & Desarrollo Alimentos, Veracruz 91897, Veracruz, Mexico. [Maria Duran-Riveroll, Lorena] CONACYT, Dept Biotecnol Marina, Ctr Invest Cient & Educ Super Ensenada, Carretera Tijuana Ensenada 3918, Ensenada 22860, Baja California, Mexico. 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The open sea was significantly enriched with Prochlorococcus and Candidatus pelagibacter. Meanwhile, coastal biosystems were dominated by Marinobacter and Alcanivorax. Halophilic archaea Haloarcula and Haloquandratum, predominant in the coastal biosystem, were significantly (p < 0.05) enriched in coastal biosystems compared to the open sea. Analysis of whole genomes (n = 23,540), retrieved from EzBioCloud, detected crtI in 64.66% of genomes, while cruF was observed in 1.69% Bacteria and 40.75% Archaea. We further confirmed the viability and carotenoid pigment production by pure culture isolation (n = 1351) of extreme halophiles from sediments (n = 410 x 3) sampling at the Arabian coastline of India. All red-pigmented isolates were represented exclusively by Haloferax, resistant to saturated NaCl (6 M), and had >60% G + C content. Multidrug resistance to tetracycline, gentamicin, ampicillin, and chloramphenicol were also observed. 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From all molecular tools available today to resolve the dynamics of microbial communities, metaproteomics stands out, allowing the establishment of phenotype-genotype linkages. Despite its rapid development, this technology has faced many technical challenges that still hamper its potential power. How to maximize the number of protein identification, improve quality of protein annotation, and provide reliable ecological interpretation are questions of immediate urgency. In our study, we used a robust metaproteomic workflow combining two protein fractionation approaches (gel-based versus gel-free) and four protein search databases derived from the same metagenome to analyze the same seawater sample. The resulting eight metaproteomes provided different outcomes in terms of (i) total protein numbers, (ii) taxonomic structures, and (iii) protein functions. 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Here, we analyzed the core genomes of 16 phylogenetically diverse and ecologically relevant bacterioplankton lineages, each consisting of up to five genomes varying at the strain level. Statistical approaches identified from each lineage up to similar to 50 loci showing anomalously high divergence at synonymous sites, which is best explained by recombination with distantly related organisms. The enriched gene categories in these outlier loci match well with the characteristics previously identified as the key phenotypes of these lineages. Examples are antibiotic synthesis and detoxification in Phaeobacter inhibens, exopolysaccharide production in Alteromonas macleodii, hydrocarbon degradation in Marinobacter hydrocarbonoclasticus, and cold adaptation in Pseudo-alteromonas haloplanktis. Intriguingly, the outlier loci feature polysaccharide catabolism in Cellulophaga baltica but not in Cellulophaga lytica, consistent with their primary habitat preferences in macroalgae and beach sands, respectively. Likewise, analysis of Prochlorococcus showed that photosynthesis-related genes listed in the outlier loci are found only in the high-light-adapted ecotype and not in the low-light adapted ecotype. These observations strongly suggest that recombination with distant relatives is a key mechanism driving the ecological diversification among marine bacterial lineages. IMPORTANCE Acquisition of new metabolic genes has been known as an important mechanism driving bacterial evolution and adaptation in the ocean, but acquisition of novel alleles of existing genes and its potential ecological role have not been examined. 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Brewster, Jodi L. Hall, Kelsi R. Evans, Gary B. Patrick, Wayne M. TI Primordial-like enzymes from bacteria with reduced genomes SO MOLECULAR MICROBIOLOGY DT Article AB The first cells probably possessed rudimentary metabolic networks, built using a handful of multifunctional enzymes. The promiscuous activities of modern enzymes are often assumed to be relics of this primordial era; however, by definition these activities are no longer physiological. There are many fewer examples of enzymes using a single active site to catalyze multiple physiologically-relevant reactions. Previously, we characterized the promiscuous alanine racemase (ALR) activity of Escherichia coli cystathionine -lyase (CBL). Now we have discovered that several bacteria with reduced genomes lack alr, but contain metC (encoding CBL). We characterized the CBL enzymes from three of these: Pelagibacter ubique, the Wolbachia endosymbiont of Drosophila melanogaster (wMel) and Thermotoga maritima. Each is a multifunctional CBL/ALR. However, we also show that CBL activity is no longer required in these bacteria. Instead, the wMel and T. maritima enzymes are physiologically bi-functional alanine/glutamate racemases. They are not highly active, but they are clearly sufficient. Given the abundance of the microorganisms using them, we suggest that much of the planet's biochemistry is carried out by enzymes that are quite different from the highly-active exemplars usually found in textbooks. Instead, primordial-like enzymes may be an essential part of the adaptive strategy associated with streamlining. C1 [Ferla, Matteo P.; Brewster, Jodi L.; Hall, Kelsi R.; Patrick, Wayne M.] Univ Otago, Dept Biochem, Dunedin, New Zealand. [Evans, Gary B.] Victoria Univ, Ferrier Res Inst, Lower Hutt, New Zealand. [Ferla, Matteo P.] Univ Oxford, Dept Biochem, Oxford, England. RP Patrick, WM (corresponding author), Univ Otago, Dept Biochem, Dunedin, New Zealand. 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Carrion, Ornella Johnston, Andrew W. B. Murrell, J. Colin Liu, Ji Zheng, Yanfen Zhang, Xiao-Hua Todd, Jonathan D. TI Oceanospirillales containing the DMSP lyase DddD are key utilisers of carbon from DMSP in coastal seawater SO MICROBIOME DT Article AB Background: Ubiquitous and diverse marine microorganisms utilise the abundant organosulfur molecule dimethylsulfoniopropionate (DMSP), the main precursor of the climate-active gas dimethylsulfide (DMS), as a source of carbon, sulfur and/or signalling molecules. However, it is currently difficult to discern which microbes actively catabolise DMSP in the environment, why they do so and the pathways used. Results: Here, a novel DNA-stable isotope probing (SIP) approach, where only the propionate and not the DMS moiety of DMSP was C-13-labelled, was strategically applied to identify key microorganisms actively using DMSP and also likely DMS as a carbon source, and their catabolic enzymes, in North Sea water. Metagenomic analysis of natural seawater suggested that Rhodobacterales (Roseobacter group) and SAR11 bacteria were the major microorganisms degrading DMSP via demethylation and, to a lesser extent, DddP-driven DMSP lysis pathways. However, neither Rhodobacterales and SAR11 bacteria nor their DMSP catabolic genes were prominently labelled in DNA-SIP experiments, suggesting they use DMSP as a sulfur source and/or in signalling pathways, and not primarily for carbon requirements. Instead, DNA-SIP identified gammaproteobacterial Oceanospirillales, e.g. Amphritea, and their DMSP lyase DddD as the dominant microorganisms/enzymes using DMSP as a carbon source. Supporting this, most gammaproteobacterial (with DddD) but few alphaproteobacterial seawater isolates grew on DMSP as sole carbon source and produced DMS. Furthermore, our DNA-SIP strategy also identified Methylophaga and other Piscirickettsiaceae as key bacteria likely using the DMS, generated from DMSP lysis, as a carbon source. 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Peng, Vivian Tyler, Charlotte Wagh, Protima TI Marine bacterioplankton biomass, activity and community structure in the vicinity of Antarctic icebergs SO DEEP-SEA RESEARCH PART II-TOPICAL STUDIES IN OCEANOGRAPHY DT Article AB We studied marine bacterioplankton in the Scotia Sea in June 2008 and in the northwest Weddell Sea in March to mid April 2009 in waters proximal to three free-drifting icebergs (SS-1, A-43k, and C-18a), in a region with a high density of smaller icebergs (iceberg alley), and at stations that were upstream of the iceberg trajectories designated as far-field reference sites that were between 16-75 km away. Hydrographic parameters were used to define water masses in which comparisons between bacterioplankton-associated characteristics (abundance, leucine incorporation into protein, aminopeptidase activities and community structure) within and between water masses could be made. Early winter Scotia Sea bacterioplankton had low levels of cells and low heterotrophic production rates in the upper 50 m. Influences of the icebergs on bacterioplankton at this time of year were minimal, if not deleterious, as we found lower levels of heterotrophic production near A-43k in comparison to stations > 16 km away. Additionally, the results point to small but significant differences in cell abundance, heterotrophic production, and community structure between the two icebergs studied. These icebergs differed greatly in size and the findings suggest that the larger iceberg had a greater effect. In the NW Weddell Sea in March-mid April bacterioplankton were twice as abundant and had heterotrophic productions rates that were 8-fold higher than what we determined in the Scotia Sea, though levels were still quite low, which is typical for autumn. We did not detect direct iceberg-related influences on the bacterioplankton characteristics studied here. Clues to understanding bacterioplankton responses may lie in the details of community structure, as there were some significant differences in community structure in the winter water and underlying upper circumpolar deep-water masses between stations occupied close to C-18a and at stations 18 km away (i.e. Polaribacter and Pelagibacter-related 16S rRNA gene fragments were at low levels at the 18 km stations), though higher resolution, high throughput profiling tools will be needed to pinpoint specific organisms and ecological types. Likewise, a better understanding of local to regional scale structure of bacterioplankton communities is necessary. The relationship between bacterioplankton abundance and heterotrophic production suggested bottom up processes were controlling bacterioplankton during March - mid-April in the NW Weddell Sea, and that variation in temperature may play a role in substrate utilization. Trophic linkages seen between phytoplankton and zooplankton were mirrored by tight coupling between primary production and heterotrophic production in the waters immediately surrounding C-18a and in iceberg alley (but not at stations 18 and 75 km away), suggesting that indirect effects on bacterioplankton may also be important. Overall, the results show that bacterioplankton, dominated by Rhodobacteracae Pelagibacter, and uncultivated Gammaproteobacteria groups were minimally influenced by icebergs in the regions and seasons studied here - at least directly - though further work addressing different scales, sizes of icebergs, and seasons is needed to better understand bacterioplankton-associated ecological processes and carbon cycling in regions of high iceberg production. (C) 2010 Elsevier Ltd. All rights reserved. C1 [Murray, Alison E.; Peng, Vivian; Tyler, Charlotte; Wagh, Protima] Desert Res Inst, Reno, NV 89512 USA. 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Miller, Samuel E. Kroll, Kourtney L. Willis, Amy D. Rappe, Michael S. Pan, Tao Eren, A. Murat TI Structure-informed microbial population genetics elucidate selective pressures that shape protein evolution SO SCIENCE ADVANCES DT Article AB Comprehensive sampling of natural genetic diversity with metagenomics enables highly resolved insights into the interplay between ecology and evolution. However, resolving adaptive, neutral, or purifying processes of evolution from intrapopulation genomic variation remains a challenge, partly due to the sole reliance on gene sequences to interpret variants. Here, we describe an approach to analyze genetic variation in the context of predicted protein structures and apply it to a marine microbial population within the SAR11 subclade 1a.3.V, which dominates low-latitude surface oceans. Our analyses reveal a tight association between genetic variation and protein structure. 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Murat] Carl von Ossietzky Univ Oldenburg, Inst Chem & Biol Marine Environm, Oldenburg, Germany. [Eren, A. Murat] Alfred Wegener Inst Polar & Marine Res, Bremerhaven, Germany. [Eren, A. Murat] Helmholtz Inst Funct Marine Biodivers, Oldenburg, Germany. RP Kiefl, E; Eren, AM (corresponding author), Univ Chicago, Dept Med, Chicago, IL 60637 USA.; Kiefl, E (corresponding author), Univ Chicago, Grad Program Biophys Sci, Chicago, IL 60637 USA.; Eren, AM (corresponding author), Marine Biol Lab, Josephine Bay Paul Ctr Comparat Mol Biol & Evolut, Woods Hole, MA 02543 USA.; Eren, AM (corresponding author), Carl von Ossietzky Univ Oldenburg, Inst Chem & Biol Marine Environm, Oldenburg, Germany.; Eren, AM (corresponding author), Alfred Wegener Inst Polar & Marine Res, Bremerhaven, Germany.; Eren, AM (corresponding author), Helmholtz Inst Funct Marine Biodivers, Oldenburg, Germany. 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Fusion proteins from marine alpha-proteobacterium Candidatus Pelagibacter ubique, actinobacterium Nocardia farcinica, chlorobi species Chloroherpeton thalassium, and beta-proteobacterium D. acidovorans each produce a different profile of non-native polyamines including sym-norspermidine when expressed in Escherichia coli. The different aminopropyltransferase activities together with phylogenetic analysis confirm independent evolutionary origins for some fusions. Comparative genomic analysis strongly indicates that gene fusions arose by merger of adjacent open reading frames. Independent fusion events, and horizontal and vertical gene transfer contributed to the scattered phyletic distribution of the gene fusions. Surprisingly, expression of fusion genes in E. coli and S. Typhimurium revealed novel latent spermidine catabolic activity producing non-native 1,3-diaminopropane in these species. We have also identified fusions of polyamine biosynthetic enzymes agmatine deiminase and N-carbamoylputrescine amidohydrolase in archaea, and of S-adenosylmethionine decarboxylase and ornithine decarboxylase in the single-celled green alga Micromonas. C1 [Michael, Anthony J.] Univ Texas SW Med Ctr Dallas, Dept Pharmacol, Dallas, TX 75390 USA. [Green, Robert; Hanfrey, Colin C.; Elliott, Katherine A.] Inst Food Res, Norwich NR4 7UA, Norfolk, England. [McCloskey, Diane E.; Wang, Xiaojing; Kanugula, Sreenivas; Pegg, Anthony E.] Penn State Univ, Coll Med, Dept Cellular & Mol Physiol, Milton S Hershey Med Ctr, Hershey, PA 17033 USA. RP Michael, AJ (corresponding author), Univ Texas SW Med Ctr Dallas, Dept Pharmacol, Dallas, TX 75390 USA. 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Bertolino, Paola Vershinin, Konstantin E. Lopez-Garcia, Purificacion TI Marine signature taxa and core microbial community stability along latitudinal and vertical gradients in sediments of the deepest freshwater lake SO ISME JOURNAL DT Article AB Lake Baikal is the deepest (~1.6 km) and most voluminous freshwater reservoir on Earth. Compared to plankton, its benthos remains poorly explored. Here, we ask whether latitude and/or depth determine benthic microbial community structure and how Baikal communities compare to those of other freshwater, brackish and marine sediments. To answer, we collected sediment upper layers (0-1 cm) across a ~600 km North-South transect covering the three basins of the lake and from littoral to bathybenthic depths (0.5-1450 m). Analysis of 16S and 18S rRNA gene amplicon sequences revealed communities with high richness and evenness where rare operational taxonomic units (OTUs) collectively dominated. Archaea represented up to 25% or prokaryotic sequences. Baikal sediments harbored typically marine eukaryotic and prokaryotic OTUs recently identified in some lakes (diplonemids, Bolidophyceae, Mamiellales, SAR202, marine-like Synechococcus, Pelagibacterales) but also SAR324, Syndiniales and Radiolaria. We hypothesize that, beyond the salinity barrier, adaptation to oligotrophy explains the presence of these otherwise typically marine lineages. Baikal core benthic communities were relatively stable across sites and seemed not determined by depth or latitude. Comparative analyses with other freshwater, brackish and marine prokaryotic sediment communities confirmed the distinctness of Baikal benthos, which include elements of similarity to marine and hydrothermally influenced systems. C1 [Reboul, Guillaume; Moreira, David; Bertolino, Paola; Lopez-Garcia, Purificacion] Univ Paris Saclay, Ctr Natl Rech Sci CNRS, Ecol Systemat Evolut, AgroParisTech, Orsay, France. 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Westrich, Jason R. Xian, Peng Holmes, Christopher D. Landing, William M. Lipp, Erin K. Wetz, Michael S. Ottesen, Elizabeth A. TI Saharan dust deposition initiates successional patterns among marine microbes in the Western Atlantic SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Deposition of aerosolized desert dust can affect marine microbial community structure and function through pulsed addition of limiting micro- and macronutrients. However, few studies have captured responses to dust deposition in situ following trans-oceanic transport. We conducted a 26-d time series evaluating biogeochemical and microbial community response to Saharan dust deposition in surface waters in the subtropical western Atlantic (Florida Keys National Marine Sanctuary, U.S.A.). Following periods of elevated atmospheric dust concentrations, particulate and dissolved iron concentrations increased in surface waters. 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Overall, proteomic SIP facilitated quantitative comparisons of DFAA assimilation by specific taxa, both between sympatric populations and between protein functional groups within discrete populations, allowing an unprecedented examination of population level metabolic responses to resource acquisition in complex microbial communities. IMPORTANCE An estimated 50 gigatons of carbon is annually fixed within marine systems, of which heterotrophic microbial populations process nearly half. These communities vary in composition and activity across spatial and temporal scales, so understanding how these changes affect global processes requires the delineation of functional roles for individual members. 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In the present study, bacterial biogeography in the Kongsfjorden seawaters showed distinct habitat patterns according to water mass classification and habitat transition between Atlantic and fjord surface waters. Moreover, we estimated phylogenetic diversity of bacterial communities using the net relatedness, nearest taxon, and beta nearest taxon indices. We found the influence of freshwater input from glacier melting in shaping bacterial assemblage composition through the stochastic model. We further evaluated bacterial contributions to phytoplankton-derived dimethylsulfoniopropionate (DMSP) using a quantitative PCR (qPCR) measurement with demethylation (dmdA) and cleavage (dddP) genes of two fundamentally different processes. Our qPCR results imply that bacterial DMSP degradation follows the Atlantic inflow during summer in Kongsfjorden. These findings suggest that the Atlantic inflow and glacial melting influence bacterial community composition and assembly processes and thus affect the degradation of phytoplankton-derived organic matter in an Arctic fjord. C1 [Han, Dukki; Unno, Tatsuya] Jeju Natl Univ, Jeju 63243, Jeju Special Se, South Korea. [Han, Dukki; Richter-Heitmann, Tim] Univ Bremen, Fac Biol Chem, Microbial Ecophysiol Grp, Bremen, Germany. [Kim, Il-Nam] Incheon Natl Univ, Dept Marine Sci, Incheon 22012, South Korea. [Choy, Eunjung; Park, Ki-Tae; Nam, Seung-Il] Korea Polar Res Inst, Incheon 21990, South Korea. [Kim, Jungman] Jeju Natl Univ, Res Inst Basic Sci, Jeju 63243, Jeju Special Se, South Korea. RP Han, D (corresponding author), Jeju Natl Univ, Jeju 63243, Jeju Special Se, South Korea.; Han, D (corresponding author), Univ Bremen, Fac Biol Chem, Microbial Ecophysiol Grp, Bremen, Germany.; Nam, SI (corresponding author), Korea Polar Res Inst, Incheon 21990, South Korea. 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Functional redundancy and metabolic tuning are two mechanisms that have been proposed to shape microbial response to environmental forcing. However, little is known about their roles in the oligotrophic surface ocean due to less integrative characterization of community taxonomy and function. Here, we applied an integrated meta-omics-based approach, from genes to proteins, to investigate the microbial community of the oligotrophic northern Indian Ocean. Insignificant spatial variabilities of both genomic and proteomic compositions indicated a stable microbial community that was dominated by Prochlorococcus, Synechococcus, and SAR11. However, fine tuning of some metabolic functions that are mainly driven by salinity and temperature was observed. Intriguingly, a tuning divergence occurred between metabolic potential and activity in response to different environmental perturbations. 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TI Temporal and spatial dynamics of Bacteria, Archaea and protists in equatorial coastal waters SO SCIENTIFIC REPORTS DT Article AB Singapore, an equatorial island in South East Asia, is influenced by a bi-annual reversal of wind directions which defines two monsoon seasons. We characterized the dynamics of the microbial communities of Singapore coastal waters by collecting monthly samples between February 2017 and July 2018 at four sites located across two straits with different trophic status, and sequencing the V6-V8 region of the small sub-unit ribosomal RNA gene (rRNA gene) of Bacteria, Archaea, and Eukaryota. Johor Strait, which is subjected to wider environmental fluctuations from anthropogenic activities, presented a higher abundance of copiotrophic microbes, including Cellvibrionales and Rhodobacterales. 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Chisholm, Sallie W. TI Ecosystem-specific selection pressures revealed through comparative population genomics SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Bacterial populations harbor vast genetic diversity that is continually shaped by abiotic and biotic selective pressures, as well as by neutral processes. Individuals coexisting in the same geographically defined population often have significantly different gene content, but whether this variation is largely adaptive or neutral remains poorly understood. Here we quantify heterogeneity in gene content for two model marine microbes, Prochlorococcus and Pelagibacter, within and between populations in the Atlantic and Pacific Oceans, to begin to understand the selective pressures that are shaping these "population genomes." We discovered a large fraction of genes that are rare in each population, reflecting continual gene transfer and loss. Despite this high variation within each population, only a few genes significantly differ in abundance between the two biogeochemically distinct environments; nearly all of these are related to phosphorus acquisition and are enriched in the Atlantic relative to the Pacific. Moreover, P-related genes from the two sites form phylogenetically distinct clusters, whereas housekeeping genes do not, consistent with a recent spread of adaptive P-related genes in the Atlantic populations. These findings implicate phosphorus availability as the dominant selective force driving divergence between these populations, and demonstrate the promise of this approach for revealing selective agents in more complex microbial systems. C1 [Coleman, Maureen L.; Chisholm, Sallie W.] MIT, Dept Civil & Environm Engn, Cambridge, MA 02139 USA. [Chisholm, Sallie W.] MIT, Dept Biol, Cambridge, MA 02139 USA. 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In this study, we employed bar-coded 454 pyrosequencing approach to investigate the bacterial community variation during a Microcystis water bloom in Guanting Reservoir, Beijing. More than 140,000 sequences were generated and assigned to 7133 operational taxonomic units (OTUs) which belong to 18 phyla through Ribosomal Database Pyrosequencing (RDP) database. Microcystis was detected at low abundance in July then broke out and became the exclusive dominant genus in September. Meanwhile, bacteria composition changed dramatically after the Microcystis water bloom reflected by the decrease of bacteria abundance, diversity and evenness indices. The classify results indicated that dominant species in July were Pelagibacter, Haliscomenobacter, Rhodobacter and Fluviicola. In September, Methylotenera, Flavobacterium and Methylophilus were the dominant genera. It is worth noticing that all the dominant genera in July was more or less related to nitrogen and phosphorus cycling while in September, dominant genera Flavobacterium was reported as Microcystis lysing bacterium and may be a symptom of the coming postbloom phase of Microcystis water bloom in Guanting Reservoir. (C) 2010 Published by Elsevier Ltd. Selection and/or peer-review under responsibility of Society for Resources, Environment and Engineering C1 [Chen Cheng; Ding Aizhong; Sun Yujiao] Beijing Normal Univ, Coll Water Sci, 19 Xinjiekouwai St, Beijing 100875, Peoples R China. [Zhang Zaichao; Wu Jiayan; Xiao Jingfa] Chinese Acad Sci, Beijing Inst Genom, Key Lab Genome Sci & Informat, CAS, Beijing 100029, Peoples R China. RP Chen, C (corresponding author), Beijing Normal Univ, Coll Water Sci, 19 Xinjiekouwai St, Beijing 100875, Peoples R China. 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In general, these microbiome disturbances were not clearly linked to specific environmental factors and responsive taxa largely differed, aside from SAR11, which generally declined. However, several disturbance metagenomes identified increased phage-associated genes, suggesting that unexplained community shifts might be caused by increased mortality. Furthermore, a category 1 hurricane, the only event that would likely be classified a priori as an environmental disturbance, was not an outlier in microbiome composition, but did enhance a bloom in seasonally abundant phytoplankton. Thus, as extreme environmental changes intensify, assumptions of what constitutes a disturbance should be re-examined in the context of ecological history and microbiome responses. C1 [Gronniger, Jessica L.; Wang, Zhao; Ward, Christopher S.; Mu, Han; Gu, Junyao; Johnson, Zackary I.; Hunt, Dana E.] Duke Univ, Marine Lab, Beaufort, NC 28516 USA. 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TI Seasonal patterns in the sunlight sensitivity of bacterioplankton from Mediterranean surface coastal waters SO FEMS MICROBIOLOGY ECOLOGY DT Article AB The sensitivity of coastal marine bacterioplankton to natural photosynthetically active radiation (PAR, 400700 nm) and ultraviolet radiation (UVR, 280400 nm) was evaluated in five experiments over a seasonal cycle in the Blanes Bay, NW Mediterranean Sea. Exposure to natural solar radiation generally inhibited bulk bacterial activities or damaged membrane integrity when irradiances were high (i.e. spring and summer experiments) and, in general, UVB (280320 nm) accounted for most of the inhibition. When assessing activity (3H-leucine uptake) at the single-cell level by microautoradiography and rRNA gene probing, seasonally varying responses and sensitivities were found among bacterial groups. While autumn and winter irradiances seemed too low to cause changes in activity, variable effects were found in spring and summer. 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TI Rhythmicity of coastal marine picoeukaryotes, bacteria and archaea despite irregular environmental perturbations SO ISME JOURNAL DT Article AB Seasonality in marine microorganisms has been classically observed in phytoplankton blooms, and more recently studied at the community level in prokaryotes, but rarely investigated at the scale of individual microbial taxa. Here we test if specific marine eukaryotic phytoplankton, bacterial and archaeal taxa display yearly rhythms at a coastal site impacted by irregular environmental perturbations. Our seven-year study in the Bay of Banyuls (North Western Mediterranean Sea) shows that despite some fluctuating environmental conditions, many microbial taxa displayed significant yearly rhythms. The robust rhythmicity was found in both autotrophs (picoeukaryotes and cyanobacteria) and heterotrophic prokaryotes. Sporadic meteorological events and irregular nutrient supplies did, however, trigger the appearance of less common non-rhythmic taxa. 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Parajka, Juraj Reischer, Georg H. Stadler, Philipp Blaschke, Alfred P. Bloeschl, Guenter Mach, Robert L. Kirschner, Alexander K. T. Farnleitner, Andreas H. Eiler, Alexander TI Bacterial diversity along a 2600 km river continuum SO ENVIRONMENTAL MICROBIOLOGY DT Article AB The bacterioplankton diversity in large rivers has thus far been under-sampled despite the importance of streams and rivers as components of continental landscapes. Here, we present a comprehensive dataset detailing the bacterioplankton diversity along the midstream of the Danube River and its tributaries. Using 16S rRNA-gene amplicon sequencing, our analysis revealed that bacterial richness and evenness gradually declined downriver in both the free-living and particle-associated bacterial communities. These shifts were also supported by beta diversity analysis, where the effects of tributaries were negligible in regards to the overall variation. 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Here, we applied CARD-FISH to quantify bacterial community structure from the subtropics to Antarctica between 10 degrees W and 10 degrees E, covering four biogeographic provinces with distinct environmental properties. In addition, incorporation of radiolabeled glucose, amino acids and leucine via MAR-FISH served to quantify the contribution to substrate turnover by selected bacterial groups. SAR11, Bacteroidetes, Gammaproteobacteria and the Roseobacter group accounted for the majority of the bacterial community (52%-88% of DAPI-stained cells) but showed little distributional variation between provinces. In contrast, taxonomic subclades Polaribacter, NS5, NS2b (Bacteroidetes) as well as RCA (Roseobacter group) featured marked geographic variation, illustrated by NMDS and coefficients of variation. Roseobacter (specifically RCA) and Gammaproteobacteria constituted considerable fractions of cells incorporating glucose and amino acids respectively. 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Zhao, Yanlin Chen, Yin TI Lipidomic Analysis of Roseobacters of the Pelagic RCA Cluster and Their Response to Phosphorus Limitation SO FRONTIERS IN MICROBIOLOGY DT Article AB The marine roseobacter-clade affiliated cluster (RCA) represents one of the most abundant groups of bacterioplankton in the global oceans, particularly in temperate and sub-polar regions. They play a key role in the biogeochemical cycling of various elements and are important players in oceanic climate-active trace gas metabolism. In contrast to copiotrophic roseobacter counterparts such as Ruegeria pomeroyi DSS-3 and Phaeobacter sp. MED193, RCA bacteria are truly pelagic and have smaller genomes. We have previously shown that RCA bacteria do not appear to encode the PlcP-mediated lipid remodeling pathway, whereby marine heterotrophic bacteria remodel their membrane lipid composition in response to phosphorus (P) stress by substituting membrane glycerophospholipids with alternative glycolipids or betaine lipids. 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Sintes, Eva TI Microbes mediating the sulfur cycle in the Atlantic Ocean and their link to chemolithoautotrophy SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Only about 10%-30% of the organic matter produced in the epipelagic layers reaches the dark ocean. Under these limiting conditions, reduced inorganic substrates might be used as an energy source to fuel prokaryotic chemoautotrophic and/or mixotrophic activity. The aprA gene encodes the alpha subunit of the adenosine-5 '-phosphosulfate (APS) reductase, present in sulfate-reducing (SRP) and sulfur-oxidizing prokaryotes (SOP). The sulfur-oxidizing pathway can be coupled to inorganic carbon fixation via the Calvin-Benson-Bassham cycle. The abundances of aprA and cbbM, encoding RuBisCO form II (the key CO2 fixing enzyme), were determined over the entire water column along a latitudinal transect in the Atlantic from 64 degrees N to 50 degrees S covering six oceanic provinces. 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DeBruyn, Jennifer M. Maas, Elizabeth W. Boyd, Philip W. Wilhelm, Steven W. TI Temporal changes in particle-associated microbial communities after interception by nonlethal sediment traps SO FEMS MICROBIOLOGY ECOLOGY DT Article AB Using marine sediment traps (named RESPIRE for REspiration of Sinking Particles In the subsuRface ocEan) designed to collect sinking particles and associated microbial communities in situ, we collected and incubated marine aggregates/particles in the southern Pacific Ocean from separate phytoplankton bloom events in situ. We determined the phylogenetic affiliation for the microorganisms growing on aggregates by pyrosequencing partial 16S rRNA gene amplicons. Water column samples were also collected and sequenced for comparison between sinking-particle-associated and planktonic bacterial communities. Statistically significant differences were found between the water column and sediment trap bacteria. 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Groundwater samples contained high concentrations of acetate, oxalate, nitrate, and sulfate, as well as radionuclides. High-throughput sequencing and analysis of the clone libraries revealed lower microbial diversity in the most strongly contaminated groundwater and a predominance of bacteria of the genera Polynucleobacter, Pusillimonas, Candidatus Pelagibacter, and of the candidate phylum Parcubacteria; these groups include species with an ultra small cell size. Archaeal sequences in the libraries belonged to ammonium oxidizers of the phylum Thaumarchaeota and methanogens of the phylum Euryarchaeota. Pure cultures of obligate and facultative ultramicrobacteria belonging to the genera Chryseobacterium, Microbacterium, Salinibacterium, Pusillimonas, Roseomonas, and Janibacter were isolated from water samples. In genomes of Pusillimonas and Roseomonas strains the genes associated with nitrate reduction, resistance to heavy metals and metalloids were revealed. 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Ravasi, Timothy Hentschel, Ute TI Revealing microbial functional activities in the Red Sea sponge Stylissa carteri by metatranscriptomics SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Sponges are important components of marine benthic environments and are associated with microbial symbionts that carry out ecologically relevant functions. Stylissa carteri is an abundant, low-microbial abundance species in the Red Sea. We aimed to achieve the functional and taxonomic characterization of the most actively expressed prokaryotic genes in S.carteri. Prokaryotic mRNA was enriched from sponge total RNA, sequenced using Illumina HiSeq technology and annotated using the metagenomics Rapid Annotation using Subsystem Technology (MG-RAST) pipeline. We detected high expression of archaeal ammonia oxidation and photosynthetic carbon fixation by members of the genus Synechococcus. Functions related to stress response and membrane transporters were among the most highly expressed by S.carteri symbionts. 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In this study, the structures and functional potentials of microbial communities associated with particles or free-living in water samples from the Pearl River Estuary were investigated using 16S rRNA gene sequencing and GeoChip 5.0 analysis. Significant differences in the community structure and genetic functional potentials between particle-associated bacteria and free-living bacteria were observed across all eight sampling sites. In particle-associated bacteria communities, Rhodobacteraceae and Flavobacteriaceae were more abundant, while SAR11 clade and SAR86 clade were the most abundant in free-living bacteria communities. The richness and abundance of functional genes involved in nutrient cycling and stress response, including carbon degradation, nitrogen fixation, DMSP degradation, and polyphosphate degradation, were much higher in particle-associated bacteria compared with free-living bacteria. Thus, the particle-associated bacteria seem to play a much more important role in the biogeochemical cycles than free-living bacteria. In conclusion, the results from this study highlight the central role played by particle-associated bacteria in structuring microbial assemblages, and their importance for mediating biogeochemical cycling in the estuarine ecosystem. C1 [Liu, Yanyang; Lin, Qun; Feng, Jiarong; Yang, Fumin; Du, Hong; Hu, Zhong; Wang, Hui] Shantou Univ, Coll Sci, Biol Dept, Shantou 515063, Peoples R China. [Liu, Yanyang; Lin, Qun; Feng, Jiarong; Yang, Fumin; Du, Hong; Hu, Zhong; Wang, Hui] Shantou Univ, Inst Marine Sci, Shantou 515063, Peoples R China. [Liu, Yanyang; Lin, Qun; Feng, Jiarong; Yang, Fumin; Du, Hong; Hu, Zhong; Wang, Hui] Shantou Univ, Guangdong Prov Key Lab Marine Biotechnol, Shantou 515063, Peoples R China. RP Wang, H (corresponding author), Shantou Univ, Coll Sci, Biol Dept, Shantou 515063, Peoples R China. 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Tully, Benjamin J. Sylvan, Jason B. Cram, Jacob A. TI Quantitative microbial taxonomy across particle size, depth, and oxygen concentration SO FRONTIERS IN MICROBIOLOGY DT Article AB Introduction Marine particles form in the ocean surface sink through the water column into the deep ocean, sequestering carbon. Microorganisms inhabit and consume carbon in these particles. The East Pacific Rise (EPR) harbors both an Oxygen Deficient Zone (ODZ) and a non-buoyant plume region formed from hydrothermal vents located on the ocean floor, allowing us to explore relationships between microbial community and particle size between a range of environments. Methods In this study, we quantified microbial diversity using a fractionation method that separated particles into seven fine scale fractions (0.2-1.2, 1.2-5, 5-20, 20-53, 53-180,180-500, >500 mu m), and included a spike-in standard for sequencing the 16S rRNA gene. Size fractionated organic carbon into the same fractions enabled the calculation of bacterial 16S rRNA copies per mu g C and per liter. Results There was a large increase in the bacterial 16S rRNA copies/ug C and copies/L on particles >180 mu m between the upper water column and the deep water column. Though the total concentration of organic C in particles decreased in the deep water column, the density of bacteria on large particles increased at depth. The microbial community varied statistically significantly as a function of particle size and depth. Quantitative abundance estimates found that ostensibly obligate free-living microbes, such as SAR11 and Thaumarcheota, were more abundant in the free-living fraction but also common and abundant in the particulate size fractions. Conversely, ostensibly obligate particle attached bacteria such as members of Bacteroidetes and Planctomycetes, while most abundant on particles, were also present in the free living fraction. Total bacterial abundance, and the abundance of many taxonomic groups, increased in the ODZ region, particularly in the free-living fraction. Contrastingly, in the non-buoyant plume, there were highly abundant bacteria in the 5-20 and 20-53 mu m fractions but reduced bacteria present in the 53-180 and 180-500 mu m fractions. Conclusion Quantitative examination of microbial communities highlights the distribution of microbial taxa unburdened by compositional effects. These data are congruent with existing models which suggest high levels of exchange between particle-attached and free-living communities. C1 [Huanca-Valenzuela, Paulina; Fuchsman, Clara A.; Cram, Jacob A.] Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD 21613 USA. [Tully, Benjamin J.] Univ Southern Calif, Ctr Dark Energy Biosphere Invest, Los Angeles, CA USA. [Tully, Benjamin J.] Branchpoint Sci, Los Angeles, CA USA. [Sylvan, Jason B.] Texas A&M Univ, Dept Oceanog, College Stn, TX USA. 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The uncultured Pelagibacter virus vSAG 37-F6 discovered via single-virus genomics is one of the most abundant and cosmopolitan marine viruses; however, little is understood about its temporal variation. Here, we estimated the absolute number of infecting 37-F6 viruses in coastal bacterioplankton from the Mediterranean Sea by using a novel, feasible SYBR Green I chip-based digital PCR (SYBR dPCR) technique, not implemented before for enumerating (uncultured) microbes. Quantitative SYBR dPCR estimated 450-3480 genome copies of virus 37-F6 in cells/mL (i.e. infecting viruses) and a total of approximate to 10-400 putative infected cells/mL with a potential C release of 0.12-4.9 pg/ml in the analysed samples. Considering that virus 37-F6 is ubiquitous and abundant in all Tara samples, an enormous amount of C could be transformed by this virus through the 'viral shunt'. 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In this work, we report the structure for the apoenzyme DmdA from Pelagibacter ubique (2.1 angstrom), as well as for DmdA co-crystals soaked with substrate DMSP (1.6 angstrom) or the cofactor tetrahydrofolate (THF) (1.6 angstrom). Surprisingly, the overall fold of the DmdA is not similar to other enzymes that typically utilize the reduced form of THF and in fact is a triple domain structure similar to what has been observed for the glycine cleavage T protein or sarcosine oxidase. Specifically, while the THF binding fold appears conserved, previous biochemical studies have shown that all enzymes with a similar fold produce 5,10-methylene-THF, while DmdA catalyzes a redox-neutral methyl transfer reaction to produce 5-methyl-THF. On the basis of the findings presented herein and the available biochemical data, we outline a mechanism for a redox-neutral methyl transfer reaction that is novel to this conserved THF binding domain. 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Because arsenate and phosphate are indiscriminately incorporated by microbes, their ambient concentration ratios can be an important factor controlling microbial growth and metabolism. This study investigated the spatial distributions of arsenate and phosphate and the associated biogeochemical dynamics in the subtropical North and South Pacific Ocean. Vertical arsenate and phosphate profiles (<= 150 m) in most of the study areas showed a nutrient-type distribution where the concentrations increased below the euphotic zone. The arsenate and phosphate concentrations in the surface waters ranged from the detection limits (5 nM and 4 nM, respectively) to approximately 40 nM and 400 nM, respectively. The surface arsenate:phosphate ratios were typically lower than 1, but those in the western subtropical North Pacific (WSNP) were frequently higher than 1 due to phosphate depletion. In the WSNP surface waters, Prochlorococcus and Pelagibacter arsenic detoxification and phosphorus acquisition genes were abundant. Results of the onboard bioassays involving the addition of arsenate or phosphate to the surface water indicated that microbes throughout the study areas possessed arsenate resistance and those in the WSNP during summer were under serious phosphate limitation. Although phosphate limitation likely accelerates the relative cellular accumulation of toxic arsenate, the lowest particulate As:P ratios were observed in the summer WSNP, concurrent with the lowest dissolved organic P (DOP) concentrations and the highest alkaline phosphatase activities. These results imply that active As excretion and/or DOP utilization could alleviate As accumulation while maintaining the cellular P quota. C1 [Hashihama, Fuminori; Suwa, Shuhei; Kanda, Jota; Ehama, Makoto; Sakuraba, Ryousuke; Kinouchi, Shinko] Tokyo Univ Marine Sci & Technol, Dept Ocean Sci, Tokyo 1088477, Japan. 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Conversely, OTUs of the Gammaproteobacterial Sulfur Oxidizer-EOSA-1 (GSO-EOSA-1) complex, the phyla Bacteroidetes and Verrucomicrobia and order Rhodobacterales were characteristic of waters south of the PF. Functions enriched south of the PF included a range of transporters, sulfur reduction and histidine degradation to glutamate, while branched-chain amino acid transport, nucleic acid biosynthesis and methionine salvage were overrepresented north of the PF. The taxonomic and functional characteristics suggested a shift of primary production from cyanobacteria in the north to eukaryotic phytoplankton in the south, and reflected the different trophic statuses of the two regions. The study provides a new level of understanding about SO microbial communities, describing the contrasting taxonomic and functional characteristics of microbial assemblages either side of the PF. C1 [Wilkins, David; Lauro, Federico M.; Williams, Timothy J.; Demaere, Matthew Z.; Brown, Mark V.; Cavicchioli, Ricardo] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia. [Brown, Mark V.] Univ New S Wales, Evolut & Ecol Res Ctr, Sydney, NSW 2052, Australia. [Hoffman, Jeffrey M.; Andrews-Pfannkoch, Cynthia; Mcquaid, Jeffrey B.] J Craig Venter Inst, Rockville, MD 20850 USA. [Riddle, Martin J.] Australian Antarctic Div, Kingston, Tas 7050, Australia. [Rintoul, Stephen R.] CSIRO Marine & Atmospher Res, Hobart, Tas 7001, Australia. [Rintoul, Stephen R.] Ctr Australian Weather & Climate Res, Partnership Bur Meteorol, Hobart, Tas 7001, Australia. [Rintoul, Stephen R.] CSIRO, Hobart, Tas 7001, Australia. [Rintoul, Stephen R.] CSIRO Wealth Oceans Natl Res Flagship, Hobart, Tas 7001, Australia. [Rintoul, Stephen R.] Antarctic Climate & Ecosyst Cooperat Res Ctr, Hobart, Tas 7001, Australia. RP Cavicchioli, R (corresponding author), Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia. 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Bacterial groups related to heterotrophy, such as Oceanospirillales (Gammapoteobacteria), SAR11 (Alphapoteobacteria), Flavobacteriales (Bacteroida), and Rhodobacterales (Alphapoteobacteria), were the main representatives of the pelagic microbial community. Additionally, we detected abundant assemblages belonging to acetate-oxidizing manganese reducers, i.e., Alteromonas. No differences were observed in microbial community diversity among pelagic zones and water masses. These results provide the first insights into the picoplankton abundance, taxonomy and diversity, and ecological processes in the Rio Grande Rise of the Atlantic Ocean. This may also support draft regulations for deep-sea mining in the region. C1 [Neiva Ferreira, Juliana Correa; Bergo, Natascha M.; Tura, Pedro M.; Chuqui, Mateus Gustavo; Brandini, Frederico P.; Jovane, Luigi; Pellizari, Vivian H.] Univ Sao Paulo, Inst Oceanog, Sao Paulo, Brazil. 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Toyama, Danyelle Rinke, Raquel Souza de Oliveira, Tereza Cristina Garcia, Jose Wagner de Miranda, Fernando Pellon Henrique-Silva, Flavio TI Metagenomics of the Water Column in the Pristine Upper Course of the Amazon River SO PLOS ONE DT Article AB River water is a small percentage of the total freshwater on Earth but represents an essential resource for mankind. Microbes in rivers perform essential ecosystem roles including the mineralization of significant quantities of organic matter originating from terrestrial habitats. The Amazon river in particular is famous for its size and importance in the mobilization of both water and carbon out of its enormous basin. Here we present the first metagenomic study on the microbiota of this river. It presents many features in common with the other freshwater metagenome available (Lake Gatun in Panama) and much less similarity with marine samples. Among the microbial taxa found, the cosmopolitan freshwater acI lineage of the actinobacteria was clearly dominant. Group I Crenarchaea and the freshwater sister group of the marine SAR11 clade, LD12, were found alongside more exclusive and well known freshwater taxa such as Polynucleobacter. A metabolism-centric analysis revealed a disproportionate representation of pathways involved in heterotrophic carbon processing, as compared to those found in marine samples. In particular, these river microbes appear to be specialized in taking up and mineralizing allochthonous carbon derived from plant material. C1 [Ghai, Rohit; Rodriguez-Valera, Francisco; McMahon, Katherine D.] Univ Miguel Hernandez, Evolutionary Genom Grp, Dept Prod Vegetal & Microbiol, Alicante, Spain. [Toyama, Danyelle; Rinke, Raquel; Henrique-Silva, Flavio] Univ Fed Sao Carlos, Mol Biol Lab, Dept Genet & Evolucao, BR-13560 Sao Carlos, SP, Brazil. [Souza de Oliveira, Tereza Cristina] Univ Fed Amazonas, Manaus, AM, Brazil. 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Communities of Bacteria and Archaea in the ice, as in under-ice seawater, were dominated by SAR11 clade Alphaproteobacteria and Marine Group I Crenarchaeota, neither of which is known from later season sea ice. The bacterial ice library contained clones of Gammaproteobacteria from oligotrophic seawater clades (e.g. OM60, OM182) but no clones from gammaproteobacterial genera commonly detected in later season sea ice by similar methods (e.g. Colwellia, Psychrobacter). The only common sea ice bacterial genus detected in winter ice was Polaribacter. Overall, selection during ice formation and mortality during winter appear to play minor roles in the process of microbial succession that leads to distinctive spring and summer sea ice communities. C1 [Collins, R. Eric; Rocap, Gabrielle; Deming, Jody W.] Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. RP Collins, RE (corresponding author), Univ Washington, Sch Oceanog, Box 357940,1503 NE Boat St, Seattle, WA 98195 USA. 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A. Cottrell, Matthew T. Ducklow, Hugh W. Kirchman, David L. TI Geographic and Phylogenetic Variation in Bacterial Biovolume as Revealed by Protein and Nucleic Acid Staining SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Biovolume is an important characteristic of cells that shapes the contribution of microbes to total biomass and biogeochemical cycling. Most studies of bacterial cell volumes use DAPI (4',6'-diamidino-2-phenylindole), which stains nucleic acids and therefore only a portion of the cell. We used SYPRO Ruby protein stain combined with fluorescence in situ hybridization to examine biovolumes of bacteria in the total community, as well in phylogenetic subgroups. Protein-based volumes varied more and were consistently larger than DNA-based volumes by 3.3-fold on average. Bacterial cells were ca. 30% larger in the Arctic Ocean and Antarctic coastal waters than in temperate regimes. We hypothesized that geographic differences in the abundance of specific bacterial groups drove the observed patterns in biovolume. In support of this hypothesis, we found that Gammaproteobacteria and members of the Sphingobacteria-Flavobacteria group were larger in higher-latitude waters and that the mean volumes of both groups were larger than the mean bacterial volume in all environments tested. The mean cell size of SAR11 bacteria was larger than the mean cell size of the total bacterial community on average, although this varied. Protein staining increases the accuracy of biovolume measurements and gives insights into how the biomass of marine microbial communities varies over time and space. C1 [Straza, Tiffany R. A.; Cottrell, Matthew T.; Kirchman, David L.] Univ Delaware, Coll Marine & Earth Studies, Lewes, DE 19958 USA. [Ducklow, Hugh W.] Marine Biol Lab, Ctr Ecosyst, Woods Hole, MA 02543 USA. 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We previously demonstrated that triple antibiotic combination therapy induces remission in patients with active UC in randomized controlled trials (RCTs). Now, we investigate changes in the gut microbiota of patients who responded to the antibiotic combination therapy. Methods: Thirty-one patients with UC given ATM/AFM (amoxicillin, metronidazole, and tetracycline or fosfomycin) therapy for two weeks were enrolled in this study. The clinical conditions of these UC patients were evaluated by the partial Mayo score. The gut microbiota was compared via the metagenomic shot gun analysis of fecal samples. Results: Of the 31 patients, 16 and 8 experienced complete and partial remission, respectively, over three months in response to ATM/AFM therapy, whereas ATM/AFM showed no efficacy in 7 patients. The dysbiosis before treatment in the active stage could be associated with increased populations of Bacteroides, Parabacteroides, Rickenella, Clostridium, Flavonifractor, Pelagibacter, Bordetella, Massilia, and Piscrickettsia species. Metagenomic analysis revealed dramatic changes in the gut microbiota at an early stage, that is, just two weeks after starting ATM/AFM therapy. After treatment in the responder group, the populations of bifidobacterium and lactobacilli species were significantly increased, while the population of bacteroides decreased. Conclusions: These results suggest that metagenomic analysis demonstrated a marked change in the gut microbiota after antibiotic combination treatment. In the triple antibiotic combination therapy, remission was associated with an increase in bifidobacterium and lactobacilli species. C1 [Ohkusa, Toshifumi; Sato, Nobuhiro] Juntendo Univ, Dept Microbiota Res, Grad Sch Med, Tokyo 1130033, Japan. 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Haro-Moreno, Jose M. Rodriguez-Valera, Francisco Almagro-Moreno, Salvador Lopez-Perez, Mario TI Single-amplified genomes reveal most streamlined free-living marine bacteria SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Evolutionary adaptations of prokaryotes to the environment sometimes result in genome reduction. Our knowledge of this phenomenon among free-living bacteria remains scarce. We address the dynamics and limits of genome reduction by examining one of the most abundant bacteria in the ocean, the SAR86 clade. Despite its abundance, comparative genomics has been limited by the absence of pure cultures and the poor representation in metagenome-assembled genomes. We co-assembled multiple previously available single-amplified genomes to obtain the first complete genomes from members of the four families. All families showed a convergent evolutionary trajectory with characteristic features of streamlined genomes, most pronounced in the TMED112 family. This family has a genome size of ca. 1 Mb and only 1 bp as median intergenic distance, exceeding values found in other abundant microbes such as SAR11, OM43 and Prochlorococcus. This genomic simplification led to a reduction in the biosynthesis of essential molecules, DNA repair-related genes, and the ability to sense and respond to environmental factors, which could suggest an evolutionary dependence on other co-occurring microbes for survival (Black Queen hypothesis). Therefore, these reconstructed genomes within the SAR86 clade provide new insights into the limits of genome reduction in free-living marine bacteria. C1 [Roda-Garcia, Juan J.; Haro-Moreno, Jose M.; Rodriguez-Valera, Francisco; Lopez-Perez, Mario] Univ Miguel Hernandez, Dept Prod Vegetal & Microbiol, Evolutionary Genom Grp, Alicante, Spain. [Almagro-Moreno, Salvador] Univ Cent Florida, Burnett Sch Biomed Sci, Orlando, FL USA. 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Gene transcript data were consistent with SAR11 using GBT as a source of methyl groups to fuel the methionine cycle. Where nitrate concentrations were higher, more GBT was partitioned for lipid biosynthesis by both bacteria and eukaryotic phytoplankton. Our data highlight unexpected metabolic pathways and potential routes of microbial metabolite exchange. C1 [Boysen, Angela K.; Kumler, William; Heal, Katherine R.; Carlson, Laura T.; Groussman, Ryan D.; Armbrust, E. Virginia; Ingalls, Anitra E.] Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. [Durham, Bryndan P.; Key, Rebecca S.] Univ Florida, Genet Inst, Dept Biol, Gainesville, FL 32610 USA. [Boysen, Angela K.] Univ Chicago, Dept Geophys Sci, Chicago, IL 60637 USA. [Heal, Katherine R.] Integral Consulting, Seattle, WA 98104 USA. RP Ingalls, AE (corresponding author), Univ Washington, Sch Oceanog, Seattle, WA 98195 USA. 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Sun Kun TI Phylogenetic diversity of planktonic bacteria in the Chukchi Borderland region in summer SO ACTA OCEANOLOGICA SINICA DT Article AB Planktonic bacteria are abundant in the Chukchi Borderland region. However, little is known about their diversity and the roles of various bacteria in the ocean. Seawater samples were collected from two stations K2S and K4S where sea ice was melting obviously. The analysis of water samples with fluorescence in situ hybridization (FISH) showed that DMSP-degrading bacteria accounted for 13% of the total bacteria at the station K2S. No aerobic anoxygenic phototrophic (AAP) bacteria were detected in both samples. The bacterial communities were characterized by two 16S rRNA gene clone libraries. Sequences fell into four major lineages of the domain Bacteria, including Proteobacteria (Alpha, Beta and Gamma subclasses), Bacteroidetes, Actinobacteria and Firmicutes. No significant difference was found between the two clone libraries. SAR11 and Rhodobacteraceae clades of Alphaproteobacteria and Pseudoalteromonas of Gammaproteobacteria constituted three dominant fractions in the clone libraries. A total of 191 heterotrophic bacterial strains were isolated and 76% showed extracellular proteolytic activity. Phylogenetic analysis reveals that the isolates fell into Gammaproteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most common genus in both the bacterial isolates and protease-producing bacteria was Pseudoalteromonas. UniFrac data showed suggestive differences in bacterial communities between the Chukchi Borderland and the northern Bering Sea. C1 [Zeng Yinxin; Yu Yong; Li Huirong; He Jianfeng; Sun Kun] Polar Res Inst China, State Ocean Adm, Key Lab Polar Sci, Shanghai 200136, Peoples R China. [Zeng Yinxin] Jimei Univ, Coll Biol Engn, Xiamen 361021, Peoples R China. [Lee, Sang H.] Pusan Natl Univ, Dept Oceanog, Pusan 609735, South Korea. 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Although phytoplankton are known to shape the composition and function of associated bacterioplankton communities, the potential bacterial responses to such a decrease of diatoms are unknown. Here we explored the changes in bacterial communities and heterotrophic production during the spring bloom in four consecutive spring blooms across several sub-basins of the Baltic Sea and related them to changes in environmental variables and in phytoplankton community structure. The taxonomic structure of bacterioplankton assemblages was partially explained by salinity and temperature but also linked to the phytoplankton community. Higher carbon biomass of the diatomsAchnanthes taeniata,Skeletonema marinoi,Thalassiosira levanderi, andChaetocerosspp. was associated with more diverse bacterial communities dominated by copiotrophic bacteria (Flavobacteriia, Gammaproteobacteria, and Betaproteobacteria) and higher bacterial production. During dinoflagellate dominance, bacterial production was low and bacterial communities were dominated by Alphaproteobacteria, mainly SAR11. Our results suggest that increases in dinoflagellate abundance during the spring bloom will largely affect the structuring and functioning of the associated bacterial communities. This could decrease pelagic remineralization of organic matter and possibly affect the bacterial grazers communities. C1 [Camarena-Gomez, Maria Teresa; Piiparinen, Jonna; Lipsewers, Tobias; Spilling, Kristian] Finnish Environm Inst, Marine Res Ctr, Helsinki, Finland. [Camarena-Gomez, Maria Teresa] Univ Helsinki, Tvarminne Zool Stn, Hango, Finland. [Ruiz-Gonzalez, Clara; Logares, Ramiro] Inst Ciencies Mar ICM CSIC, Barcelona, Spain. [Sobrino, Cristina] Univ Vigo, Fac Sci, Vigo, Spain. [Spilling, Kristian] Univ Agder, Dept Nat Sci, Kristiansand, Norway. RP Camarena-Gómez, MT (corresponding author), Finnish Environm Inst, Marine Res Ctr, Helsinki, Finland.; Camarena-Gómez, MT (corresponding author), Univ Helsinki, Tvarminne Zool Stn, Hango, Finland. 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Balague, Vanessa Pedros-Alio, Carlos TI Winter-to-summer changes in the composition and single-cell activity of near-surface Arctic prokaryotes SO ENVIRONMENTAL MICROBIOLOGY DT Article AB We collected surface samples in Franklin Bay (Western Arctic) from ice-covered to ice-free conditions, to determine seasonal changes in the identity and in situ activity of the prokaryotic assemblages. Catalysed reported fluorescence in situ hybridization was used to quantify the abundance of different groups, and combined with microautoradiography to determine the fraction of active cells taking up three substrates: glucose, amino acids and ATP. In surface waters, Archaea accounted for 16% of the total cell count in winter, but decreased to almost undetectable levels in summer, when Bacteria made up 97% of the total cell count. Alphaproteobacteria were the most abundant group followed by Bacteroidetes (average of 34% and 14% of total cell counts respectively). 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Santha, Kinga Apprill, Amy TI Coral Reef Water Microbial Communities of Jardines de la Reina, Cuba SO MICROORGANISMS DT Article AB Globally, coral reef ecosystems are undergoing significant change related to climate change and anthropogenic activities. Yet, the Cuban archipelago of Jardines de la Reina (JR) has experienced fewer stressors due to its geographical remoteness and high level of conservation. This study examines the surface and benthic reef water microbial communities associated with 32 reef sites along the JR archipelago and explores the relationship between the community composition of reef microorganisms examined using bacterial and archaeal small subunit ribosomal RNA gene (16S rRNA gene) sequencing compared to geographic, conservation/protection level, environmental, physicochemical, and reef benthic and pelagic community features. 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The predominant source of oceanic DMS is the cleavage of dimethylsulfoniopropionate (DMSP), which can be produced by marine bacteria and phytoplankton. Polar oceans, which represent about one fifth of Earth's surface, contribute significantly to the global oceanic DMS sea-air flux. However, a global overview of DMS and DMSP cycling in polar oceans is still lacking and the key genes and the microbial assemblages involved in DMSP/DMS transformation remain to be fully unveiled. Results: Here, we systematically investigated the biogeographic traits of 16 key microbial enzymes involved in DMS/DMSP cycling in 60 metagenomic samples from polar waters, together with 174 metagenome and 151 metatranscriptomes from non-polar Tara Ocean dataset. Our analyses suggest that intense DMS/DMSP cycling occurs in the polar oceans. DMSP demethylase (DmdA), DMSP lyases (DddD, DddP, and DddK), and trimethylamine monooxygenase (Tmm, which oxidizes DMS to dimethylsulfoxide) were the most prevalent bacterial genes involved in global DMS/DMSP cycling. Alphaproteobacteria (Pelagibacterales) and Gammaproteobacteria appear to play prominent roles in DMS/DMSP cycling in polar oceans. The phenomenon that multiple DMS/DMSP cycling genes co-occurred in the same bacterial genome was also observed in metagenome assembled genomes (MAGs) from polar oceans. The microbial assemblages from the polar oceans were significantly correlated with water depth rather than geographic distance, suggesting the differences of habitats between surface and deep waters rather than dispersal limitation are the key factors shaping microbial assemblages involved in DMS/DMSP cycling in polar oceans. Conclusions: Overall, this study provides a global overview of the biogeographic traits of known bacterial genes involved in DMS/DMSP cycling from the Arctic and Antarctic oceans, laying a solid foundation for further studies of DMS/DMSP cycling in polar ocean microbiome at the enzymatic, metabolic, and processual levels. C1 [Teng, Zhao-Jie; Qin, Qi-Long; Li, Jian; Zhang, Yu-Zhong] Shandong Univ, State Key Lab Microbial Technol, Marine Biotechnol Res Ctr, Qingdao 266237, Peoples R China. [Zhang, Weipeng; Fu, Hui-Hui; Wang, Peng; Wang, Min; Chen, Xiu-Lan; Zhang, Yu-Zhong; Chen, Yin; Li, Chun-Yang] Ocean Univ China, Coll Marine Life Sci, Inst Adv Ocean Study, Qingdao 266003, Peoples R China. [Fu, Hui-Hui; Wang, Peng; Chen, Xiu-Lan; Zhang, Yu-Zhong; Li, Chun-Yang] Pilot Natl Lab Marine Sci & Technol Qingdao, Lab Marine Biol & Biotechnol, Qingdao 266373, Peoples R China. 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Here, we identify potential key producers and consumers of B1 and gain new insights into the dynamics of B1 cycling through measurements of B1 and vitamers (HMP: 4-amino-5-hydroxymethyl-2-methylpyrimidine, HET: 4-methyl-5-thiazoleethanol, FAMP: N-formyl-4-amino- 5-aminomethyl-2-methylpyrimidine) in the particulate and dissolved pool in a temperate coastal system. Dissolved B1 was not the primary limiting nutrient for bacterial production and was relatively stable across seasons with concentrations ranging from 74-117 pM, indicating a balance of supply and demand. However, vitamer concentration changed markedly with season as did transcripts related to vitamer salvage and transport suggesting use of vitamers by certain bacterioplankton, e.g. Pelagibacterales. Genomic and transcriptomic analyses showed that up to 78% of the bacterioplankton taxa were B1 auxotrophs. 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Nevertheless, virus-mediated taxon-specific cell lysis is few studied to date. In the present study, we investigated the taxon-specific cell lysis and estimated its contribution to the variations of community composition in rare and abundant microbial taxa. The dominant taxa Prochlorococcus, Synechococcus, SAR11, and Rhodobacteraceae displayed lower cell lysis index (CLI, the rate of extracellular to intracellular rRNA) in surface seawater. Meanwhile, Alteromonas, Pseudomonas, and Halomonas had high CLI values in the bottom seawater. Cell lysis contributed a larger percentage of variation in rare taxa (5.0%-9.4%) than in abundant taxa (0.1%-1.7%). Furthermore, linear regression analysis indicated that rare taxa were more likely to experience higher viral lysis pressure relative to abundant taxa. Our findings provide insight into the impact of virus-mediated cell lysis on prokaryotic community structure and diversity and further improve our understanding of the various abiotic and biotic factors contributing to community assembly in the ocean.IMPORTANCEVirus-induced host lysis contributes up to 40% of total prokaryotic mortality and plays crucial roles in shaping microbial composition and diversity in the ocean. Nonetheless, what taxon-specific cell lysis is caused by viruses remains to be studied. The present study, therefore, examined the taxon-specific cell lysis and estimated its contribution to the variations in the rare and abundant microbial taxa. The results demonstrate that taxon-specific mortality differed in surface and bottom of the coastal environment. In addition, active rare taxa are more susceptible to heightened lytic pressure and suggested the importance of viral lysis in regulating the microbial community composition. These results improve our understanding of bottom-up (abiotic environmental variables) and top-down (viral lysis) controls contributing to microbial community assembly in the ocean. Virus-induced host lysis contributes up to 40% of total prokaryotic mortality and plays crucial roles in shaping microbial composition and diversity in the ocean. Nonetheless, what taxon-specific cell lysis is caused by viruses remains to be studied. The present study, therefore, examined the taxon-specific cell lysis and estimated its contribution to the variations in the rare and abundant microbial taxa. The results demonstrate that taxon-specific mortality differed in surface and bottom of the coastal environment. In addition, active rare taxa are more susceptible to heightened lytic pressure and suggested the importance of viral lysis in regulating the microbial community composition. These results improve our understanding of bottom-up (abiotic environmental variables) and top-down (viral lysis) controls contributing to microbial community assembly in the ocean. C1 [Liu, Lu; Chen, Qi; Wang, Yu; Zhang, Ting; Jiao, Nianzhi; Zheng, Qiang] Xiamen Univ, Inst Marine Microbes & Ecospheres, Coll Ocean & Earth Sci, State Key Lab Marine Environm Sci, Xiamen, Peoples R China. [Liu, Lu; Chen, Qi; Wang, Yu; Zhang, Ting; Jiao, Nianzhi; Zheng, Qiang] Xiamen Univ, Fujian Key Lab Marine Carbon Sequestrat, Xiamen, Peoples R China. [Zhong, Kevin Xu] Univ British Columbia, Dept Earth Ocean & Atmospher Sci, Vancouver, BC, Canada. RP Zheng, Q (corresponding author), Xiamen Univ, Inst Marine Microbes & Ecospheres, Coll Ocean & Earth Sci, State Key Lab Marine Environm Sci, Xiamen, Peoples R China.; Zheng, Q (corresponding author), Xiamen Univ, Fujian Key Lab Marine Carbon Sequestrat, Xiamen, Peoples R China. 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Spectra attributed to the marine Roseobacter clade (MRC), the oligotrophic marine Gammaproteobacteria (OMG) group and Flavobacteria dominated in the microbial community, accounting for 21.0%, 23.2% and 12.7% of all of the detected spectra, respectively, whereas the SAR 92 clade accounted for 50% of the OMG group. The abundance of TonB-dependent receptors (TBDRs) was detected and the majority of TBDRs were attributed to the OMG, whereas a large number of ABC transporters matched to the MRC, which suggests niche separation in the microbial community. Expression of proteorhodopsin and RagB/SusD from Flavobacteria facilitates their attachment and growth on algal-derived organic matter. Taurine and glycine betaine appear to be an important source of carbon and nitrogen for the Rhodobacteraceae and SAR11 cluster. The detection of carbon monoxide dehydrogenase, formate dehydrogenase, O-acetylhomoserine sulfhydrylase and sulfur oxidation protein from the MRC demonstrated that members of the MRC play important roles in coastal ocean biogeochemical cycles. This study provides the first insight into functional processes occurring in microbial communities in coastal waters in the South China Sea. C1 [Dong, Hong-Po; Lu, Songhui] Jinan Univ, Res Ctr Harmful Algae & Marine Biol, Key Lab Eutrophicat & Red Tide Prevent, Guangdong Higher Educ Inst, Guangzhou 510632, Guangdong, Peoples R China. [Hong, Yi-Guo] Chinese Acad Sci, South China Sea Inst Oceanog, State Key Lab Trop Oceanog, Guangzhou 510632, Guangdong, Peoples R China. [Xie, Lu-Yuan] Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China. RP Dong, HP (corresponding author), Jinan Univ, Res Ctr Harmful Algae & Marine Biol, Key Lab Eutrophicat & Red Tide Prevent, Guangdong Higher Educ Inst, Guangzhou 510632, Guangdong, Peoples R China. 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Marin, Roman, III Jensen, Scott D. Feldman, Jason Birch, James M. Massion, Eugene I. DeLong, Edward F. Suzuki, Marcelino Wheeler, Kevin Scholin, Christopher A. TI Near real-time, autonomous detection of marine bacterioplankton on a coastal mooring in Monterey Bay, California, using rRNA-targeted DNA probes SO ENVIRONMENTAL MICROBIOLOGY DT Article AB A sandwich hybridization assay (SHA) was developed to detect 16S rRNAs indicative of phylogenetically distinct groups of marine bacterioplankton in a 96-well plate format as well as low-density arrays printed on a membrane support. The arrays were used in a field-deployable instrument, the Environmental Sample Processor (ESP). The SHA employs a chaotropic buffer for both cell homogenization and hybridization, thus target sequences are captured directly from crude homogenates. 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Fodelianakis, Stilianos Pitta, Paraskevi TI Responses of Free-Living Planktonic Bacterial Communities to Experimental Acidification and Warming SO MICROORGANISMS DT Article AB Climate change driven by human activities encompasses the increase in atmospheric CO2 concentration and sea-surface temperature. Little is known regarding the synergistic effects of these phenomena on bacterial communities in oligotrophic marine ecosystems that are expected to be particularly vulnerable. Here, we studied bacterial community composition changes based on 16S rRNA sequencing at two fractions (0.1-0.2 and >0.2 mu m) during a 10- day fully factorial mesocosm experiment in the eastern Mediterranean where the pH decreased by similar to 0.3 units and temperature increased by similar to 3 degrees C to project possible future changes in surface waters. 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Song, Bongkeun Anderson, Iris C. Reece, Kimberly S. TI Blooms of the harmful algae Margalefidinium polykrikoides and Alexandrium monilatum alter the York River Estuary microbiome SO HARMFUL ALGAE DT Article AB Harmful algal blooms (HABs) cause damage to fisheries, aquaculture, and human health around the globe. However, the impact of HABs on water column microbiomes and biogeochemistry is poorly understood. This study examined the impacts of consecutive blooms of the ichthyotoxic dinoflagellates Margalefidinium polykrikoides and Alexandrium monilatum on the water microbiome in the York River Estuary, Chesapeake Bay, USA. The samples dominated by single dinoflagellate species and by a mix of the two dinoflagellates had different microbiome compositions than the ones with low levels of both species. The M. polykrikoides bloom was co dominated by Winogradskyella and had increased concentrations of dissolved organic carbon. The A. monilatum bloom had little impact on the prokaryotic portion of the whole community but was associated with a specific group of prokaryotes in the particle-attached (>3 mu m) fraction including Candidatus Nitrosopumilus, Candidatus Actinomarina, SAR11 Clade Ia, Candidatus Bealeia, and Rhodobacteraceae HIMB11. Thus, blooms of these two algal species impacted the estuarine microbiome in different ways, likely leading to shifts in estuarine carbon and nutrient cycling, with M. polykrikoides potentially having a greater impact on carbon cycling in the estuarine ecosystem than A. monilatum. C1 [Fortin, Samantha G.; Song, Bongkeun; Anderson, Iris C.; Reece, Kimberly S.] Virginia Inst Marine Sci, Gloucester Point, VA 23062 USA. RP Fortin, SG; Song, B (corresponding author), Virginia Inst Marine Sci, Gloucester Point, VA 23062 USA. 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The best examples are the cyanobacterial genus Prochlorococcus, the alphaproteobacterial clade SAR11 and the gammaproteobacterial clade SAR86, which together represent over 50% of the cells in surface oceans. Several studies have identified signatures of selection on these lineages in today's ocean and have postulated selection as the primary force throughout their evolutionary history. However, massive loss of genomic DNA in these lineages often occurred in the distant past, and the selective pressures underlying these ancient events have not been assessed. Here, we probe ancient selective pressures by computing % GC-corrected rates of conservative and radical nonsynonymous nucleotide substitutions. Surprisingly, we found an excess of radical changes in several of these lineages in comparison to their relatives with larger genomes. 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The discovery of a major role of bacteria in the ocean's carbon cycle owes greatly to the development of culture-independent assemblage-level approaches; however, this should not detract from the recognition of model isolates as representing the environmental microbiome. A long-established tool for culturing bacteria, in medicine and general microbiology, has been agar plates. In addition, a great variety of liquid substrates including seawater have been used to successfully identify and cultivate important bacteria such as Pelagibacter ubique. Yet, the discrepancy between microscopic counts and plate counts, the great plate count anomaly, has led to a biased perception of the limited relevance of isolated bacteria. Linking isolates to whole-genome sequencing, phylogenetic analysis and computational modeling will result in culturable model bacteria from different habitats. 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The effect of the latter on natural bacterial assemblages was determined using ammonium amended experiments at Puyuhuapi fjord in Chilean Patagonia. Mucus was added to seawater coming from 2 and 100 m depth and ammonium, nitrite and nitrate, dissolved organic carbon (DOC), picoplankton abundance, and active composition (i-tag 16S rRNA) were followed for 24 h. The results showed a significant response from the microbial community but only at surface depth after 2 and 6 h of incubation. A reduction of DOC and ammonium concentration and accumulation of nitrite and nitrate over time was observed, mainly at 100 m. Changes in the composition of active bacteria between treatments were observed at different taxonomic levels, associated with Alphaproteobacteria (Clade SAR11), Bacteroidetes (Polaribacter) and Gammaproteobacteria (Colwellia, Oceaniserpentilla) and other bacteria such as Nitrospina sp, a nitrite-oxidizing bacteria at some hours during the incubation. 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Koplovitz, Gil Cariou, Thierry Henry, Nicolas de Vargas, Colomban Jeanthon, Christian TI Exploring the phycosphere of Emiliania huxleyi: From bloom dynamics to microbiome assembly experiments SO MOLECULAR ECOLOGY DT Article AB Coccolithophores have global ecological and biogeochemical significance as the most important calcifying marine phytoplankton group. The structure and selection of prokaryotic communities associated with the most abundant coccolithophore and bloom-forming species, Emiliania huxleyi, are still poorly known. In this study, we assessed the diversity of bacterial communities associated with an E. huxleyi bloom in the Celtic Sea (Eastern North Atlantic), exposed axenic E. huxleyi cultures to prokaryotic communities derived from bloom and non-bloom conditions, and followed the dynamics of their microbiome composition over one year. Bloom-associated prokaryotic communities were dominated by SAR11, Marine group II Euryarchaeota and Rhodobacterales and contained substantial proportions of known indicators of phytoplankton bloom demises such as Flavobacteriaceae and Pseudoalteromonadaceae. The taxonomic richness of bacteria derived from natural communities associated with axenic E. huxleyi rapidly shifted and then stabilized over time. The succession of microorganisms recruited from the environment was consistently dependent on the composition of the initial bacterioplankton community. Phycosphere-associated communities derived from the E. huxleyi bloom were highly similar to one another, suggesting deterministic processes, whereas cultures from non-bloom conditions show an effect of stochasticity. Overall, this work sheds new light on the importance of the initial inoculum composition in microbiome recruitment and elucidates the temporal dynamics of its composition and long-term stability. C1 [dos Reis, Mariana Camara; Romac, Sarah; Le Gall, Florence; Marie, Dominique; Henry, Nicolas; de Vargas, Colomban; Jeanthon, Christian] Sorbonne Univ, Ctr Natl Rech Sci, Stn Biol Roscoff, UMR7144,Adaptat & Diversit Milieu Marin, Roscoff, France. [dos Reis, Mariana Camara; Henry, Nicolas; de Vargas, Colomban; Jeanthon, Christian] Res Federat study Global Ocean Syst Ecol & Evolut, Tara GOSEE, FR2022, Paris, France. [Frada, Miguel J.; Koplovitz, Gil] Interuniv Inst Marine Sci Eilat, Elat, Israel. [Frada, Miguel J.] Hebrew Univ Jerusalem, Silberman Inst Life Sci, Dept Ecol Evolut & Behav, Jerusalem, Israel. [Cariou, Thierry] Sorbonne Univ, Ctr Natl Rech Sci, Stn Biol Roscoff, FR2424, Roscoff, France. [dos Reis, Mariana Camara] BIOCITECH, Biomillenia Design Pharmaceut, Romainville, France. [Cariou, Thierry] IRD, US191, Instrumentat Moyens Analyt Observ Geophys & Ocean, Plouzane, France. 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Vaque, Dolors Agusti, Susana Gasol, Josep M. TI Bacterial assemblage structure and carbon metabolism along a productivity gradient in the NE Atlantic Ocean SO AQUATIC MICROBIAL ECOLOGY DT Article AB Bacterioplankton have the potential to significantly affect the cycling of organic matter in the ocean; however, little is known about the linkage between bacterial assemblage structure and carbon metabolism. In this study, we investigated whether changes in the phylogenetic composition of bacterioplankton were associated with changes in bacterial carbon processing (bacterial production, respiration and biomass) in the subtropical NE Atlantic Ocean. We found consistent differences in the composition of the bacterial assemblage, as revealed by denaturing gradient gel electrophoresis (DGGE) and catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH), along a gradient from the NW African upwelling to the oligotrophic North Atlantic Subtropical Gyre. The percent contribution of Bacteroidetes, Roseobacter and Gammaproteobacteria significantly increased towards more productive waters, whereas the SAR11 clade of the Alphaproteobacteria remained relatively constant (average 28% of DAPI-stained cells) throughout the area. Changes in the composition of the bacterial assemblage detected by DGGE were weakly but significantly correlated with changes in carbon processing variables. The abundances of Roseobacter and Gammaproteobacteria were highly correlated with the concentration of particulate organic carbon and chlorophyll a, reflecting the affinity of these groups to nutrient-enriched conditions. The abundance of Roseobacter was also positively correlated with heterotrophic bacterial production, suggesting their active participation in carbon processing. C1 CSIC, Inst Ciencies Mar, Dept Biol Marina & Oceanog, Barcelona 08003, Catalunya, Spain. Univ Palmas Gran Canaria, Dept Biol, Las Palmas Gran Canaria 35017, Spain. 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William Lee, Kenneth Whyte, Lyle G. Greer, Charles W. TI Microbial community characterization of the Gully: a marine protected area SO CANADIAN JOURNAL OF MICROBIOLOGY DT Article AB The Gully is the first Fisheries and Oceans Canada marine protected area off the eastern coast of Canada. To ensure success of conservation efforts in this area, it is essential to develop a better understanding of microbial community composition from the euphotic zone to the deep sea in this previously unsurveyed environment. Denaturing gradient eel electrophoresis (DGGE) and nucleotide sequencing were used to characterize microbial community structure. DGGE results showed a clear difference in the microbial community structure between the euphotic zone and the deep sea water. Cluster analysis showed high similarity (>85%) for all the samples taken from below 500 m, but lower similarity (49%-72%) when comparing samples from above and below 500 m. 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Analysis of virus families indicated that the Myoviridae was the most common during all four seasons, and viruses in the Polyomaviridae were only present during March. Viruses in the Iridoviridae were only present during three seasons. Additionally, viruses in the Iridoviridae, Herpesviridae, and Poxviridae, which may affect fish and marine animals, appeared during different seasons. These results suggest that seasonal changes in temperature contribute to the dynamic structure of the viral community in the study area. The information presented here will be useful for comparative analyses with other marine viral communities. C1 [Hwang, Jinik; Park, So Yun; Park, Mirye; Lee, Taek-Kyun] Korea Inst Ocean Sci & Technol, South Sea Environm Res Dept, Geoje, South Korea. [Hwang, Jinik; Park, Mirye; Lee, Taek-Kyun] Korea Univ Sci & Technol, Marine Environm Sci, Daejeon, South Korea. [Lee, Sukchan] Sungkyunkwan Univ, Dept Genet Engn, Suwon, South Korea. 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Notably, the occupancy-frequency distributions were significantly bimodal with a satellite mode of rare endemic populations and a core mode of abundant cosmopolitan populations (e.g. Synechococcus, SAR11 and SAR86 clade members). Temporal changes in population distributions supported several theoretical frameworks. Still, bimodality was found among bacterioplankton communities across the entire Baltic Sea, and was also frequent in globally distributed datasets. Datasets spanning waters with widely different physicochemical characteristics or environmental gradients typically lacked significant bimodal patterns. When such datasets were divided into subsets with coherent environmental conditions, bimodal patterns emerged, highlighting the importance of positive feedbacks between local abundance and occupancy within specific biomes. 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Statistical analyses (analysis of similarities, Bray-Curtis similarity) confirmed that the microbial community structure followed closely the geochemical zonation. Yet, our characterization of the genetic potential relevant to carbon, nitrogen and sulphur cycling of each zone revealed unexpected features, including potential for facultative anaerobic methylotrophy, nitrogen fixation despite high ammonium concentrations and potential micro-aerobic nitrifiers within the chemocline. At the oxic-suboxic interface, facultative anaerobic potential was found in the widespread freshwater lineage acI (Actinobacteria), suggesting intriguing ecophysiological similarities to the marine SAR11. Evolutionary divergent lineages among diverse phyla were identified in the ancient seawater zone and may indicate novel adaptations to this unusual environment. C1 [Haas, Sebastian; Wallace, Douglas W. R.] Dalhousie Univ, Dept Oceanog, 1355 Oxford St, Halifax, NS, Canada. 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Daniel, Rolf Brinkhoff, Thorsten Simon, Meinhard TI Bacterial community dynamics during polysaccharide degradation at contrasting sites in the Southern and Atlantic Oceans SO ENVIRONMENTAL MICROBIOLOGY DT Article AB The bacterial degradation of polysaccharides is central to marine carbon cycling, but little is known about the bacterial taxa that degrade specific marine polysaccharides. Here, bacterial growth and community dynamics were studied during the degradation of the polysaccharides chitin, alginate and agarose in microcosm experiments at four contrasting locations in the Southern and Atlantic Oceans. At the Southern polar front, chitin-supplemented microcosms were characterized by higher fractions of actively growing cells and a community shift from Alphaproteobacteria to Gammaproteobacteria and Bacteroidetes. At the Antarctic ice shelf, chitin degradation was associated with growth of Bacteroidetes, with 24% higher cell numbers compared with the control. 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The autonomous sampling instrument consisted of the Environmental Sample Processor (ESP) coupled to an autonomous underwater vehicle (AUV). We tested equivalency between the ESP and traditional eDNA sampling techniques by comparing the quantification of eDNA across a broad range of taxa, from microbes (SAR11), phytoplankton (Pseudo-nitzschia spp.), and invertebrates (krill: Euphausia pacifica) to vertebrates (anchovy: Engraulis mordax). No significant differences in eDNA densities were observed between the two sample collection and filtration methods. eDNA filters collected by the ESP were preserved and stable for 21 days, the typical deployment length of the instrumentation. Finally, we demonstrated the unique capabilities of an autonomous, mobile ESP during a deployment near Monterey Bay, CA, by remotely and repeatedly sampling a water mass over 12 h. 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Chen, Lin-Xing Mori, Jiro F. Nelson, Tara E. Colenbrander Slater, Gregory F. Lindsay, Matthew B. J. Banfield, Jillian F. Warren, Lesley A. TI Alum Addition Triggers Hypoxia in an Engineered Pit Lake SO MICROORGANISMS DT Article AB Here, we examine the geobiological response to a whole-lake alum (aluminum sulfate) treatment (2016) of Base Mine Lake (BML), the first pilot-scale pit lake established in the Alberta oil sands region. The rationale for trialing this management amendment was based on its successful use to reduce internal phosphorus loading to eutrophying lakes. Modest increases in water cap epilimnetic oxygen concentrations, associated with increased Secchi depths and chlorophyll-a concentrations, were co-incident with anoxic waters immediately above the fluid fine tailings (FFT) layer post alum. Decreased water cap nitrate and detectable sulfide concentrations, as well as increased hypolimnetic phospholipid fatty acid abundances, signaled greater anaerobic heterotrophic activity. Shifts in microbial community to groups associated with greater organic carbon degradation (i.e., SAR11-LD12 subclade) and the SRB group Desulfuromonodales emerged post alum and the loss of specialist groups associated with carbon-limited, ammonia-rich restricted niches (i.e., MBAE14) also occurred. Alum treatment resulted in additional oxygen consumption associated with increased autochthonous carbon production, watercap anoxia and sulfide generation, which further exacerbate oxygen consumption associated with on-going FFT mobilized reductants. The results illustrate the importance of understanding the broader biogeochemical implications of adaptive management interventions to avoid unanticipated outcomes that pose greater risks and improve tailings reclamation for oil sands operations and, more broadly, the global mining sector. C1 [Jessen, Gerdhard L.] Univ Austral Chile, Inst Ciencias Marinas & Limnol, Valdivia 5090000, Chile. [Jessen, Gerdhard L.; Mori, Jiro F.; Nelson, Tara E. Colenbrander; Warren, Lesley A.] Univ Toronto, Dept Civil & Mineral Engn, Toronto, ON M5S 1A4, Canada. [Chen, Lin-Xing; Banfield, Jillian F.] Univ Calif Berkeley, Dept Earth & Planetary Sci, Berkeley, CA 94706 USA. [Mori, Jiro F.] Yokohama City Univ, Grad Sch Nanobiosci, Yokohama, Kanagawa 2360027, Japan. [Nelson, Tara E. Colenbrander; Slater, Gregory F.; Warren, Lesley A.] McMaster Univ, Sch Earth Environm & Soc, Hamilton, ON L8S 4L8, Canada. [Lindsay, Matthew B. J.] Univ Saskatchewan, Dept Geol Sci, Saskatoon, SK S7N 5E2, Canada. RP Jessen, GL (corresponding author), Univ Austral Chile, Inst Ciencias Marinas & Limnol, Valdivia 5090000, Chile.; Jessen, GL; Warren, LA (corresponding author), Univ Toronto, Dept Civil & Mineral Engn, Toronto, ON M5S 1A4, Canada.; Warren, LA (corresponding author), McMaster Univ, Sch Earth Environm & Soc, Hamilton, ON L8S 4L8, Canada. 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Preston, CM Chavez, FP DeLong, EF AF Suzuki, MT Preston, CM Chavez, FP DeLong, EF TI Quantitative mapping of bacterioplankton populations in seawater: field tests across an upwelling plume in Monterey Bay SO AQUATIC MICROBIAL ECOLOGY DT Article AB Few methods are available for quantifying specific prokaryotic taxa in marine plankton samples. In this study, we report a novel sampling and analysis strategy that circumvents some of the difficulties associated with current methods. This new approach allows for increased spatial, temporal and phylogenetic resolution over what has been achievable in routine bacterioplankton surveys. Picoplankton from small volume samples (30 mi) were collected on polysulfone filters and DNA was extracted with a commercially available DNA purification kit. The contribution of different bacterioplankton members at the group and subgroup levels was quantified by 5' nuclease assays. Percentages of small subunit (SSU) rDNAs from SAR11, SAR86, Roseobacter, Cytophaga and Synechococcus clades in DNA extracted from small samples were compared with SSU rDNA in DNA samples extracted from 6 to 91 seawater. Only small differences were observed between the methods. The approach was also tested by estimating gene copy numbers in a seawater sample spiked with varying numbers of cells from a cultivated marine Roseobacter strain. Finally we measured SSU rDNAs from the same groups of marine bacterioplankton in samples from a rapid survey of an upwelling plume in Monterey Bay, California, USA. A strong negative correlation between the percentage of Cytophagales and recently upwelled water, and an overlap between higher SAR86 percentages and a chlorophyll a concentration peak was found. The results confirm that rapid mapping of specific bacterioplankton groups is achievable using small samples and 5' nuclease assays. C1 Monterey Bay Aquarium Res Inst, Moss Landing, CA 95039 USA. RP DeLong, EF (corresponding author), Monterey Bay Aquarium Res Inst, 7700 Sandholdt Rd,POB 628, Moss Landing, CA 95039 USA. 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Subsequent RNA sequencing analyses yielded genome-wide transcriptome profiles of eukaryotic (Ostreococcus) and bacterial (Synechococcus) photosynthetic picoplankton as well as proteorhodopsin-containing heterotrophs, including Pelagibacter, SAR86-cluster Gammaproteobacteria, and marine Euryarchaea. The photosynthetic picoplankton exhibited strong diel rhythms over thousands of gene transcripts that were remarkably consistent with diel cycling observed in laboratory pure cultures. In contrast, the heterotrophs did not cycle diurnally. Instead, heterotrophic picoplankton populations exhibited cross-species synchronous, tightly regulated, temporally variable patterns of gene expression for many genes, particularly those genes associated with growth and nutrient acquisition. This multitaxon, population-wide gene regulation seemed to reflect sporadic, short-term, reversible responses to high-frequency environmental variability. 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Carlson, Craig A. TI Tracking differential incorporation of dissolved organic carbon types among diverse lineages of Sargasso Sea bacterioplankton SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Bacterioplankton are the primary trophic conduit for dissolved organic carbon (DOC) and linking community structure with DOC utilization is central to understanding global carbon cycling. We coupled stable isotope probing (SIP) with 16S rRNA pyrosequencing in dark seawater culture experiments on euphotic and mesopelagic communities from the Sargasso Sea. Parallel cultures were amended with equimolar quantities of four DO13C substrates to simultaneously evaluate community utilization and population-specific incorporation. 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Liu, Zhanfei Erdner, Deana L. TI Natural Sunlight Shapes Crude Oil-Degrading Bacterial Communities in Northern Gulf of Mexico Surface Waters SO FRONTIERS IN MICROBIOLOGY DT Article AB Following the Deepwater Horizon (DWH) spill in 2010, an enormous amount of oil was observed in the deep and surface waters of the northern Gulf of Mexico. Surface waters are characterized by intense sunlight and high temperature during summer. While the oil-degrading bacterial communities in the deep-sea plume have been widely investigated, the effect of natural sunlight on those in oil polluted surface waters remains unexplored to date. In this study, we incubated surface water from the DWH site with amendments of crude oil, Corexit dispersant, or both for 36 days under natural sunlight in the northern Gulf of Mexico. The bacterial community was analyzed over time for total abundance, density of alkane and polycyclic aromatic hydrocarbon degraders, and community composition via pyrosequencing. Our results showed that, for treatments with oil and/or Corexit, sunlight significantly reduced bacterial diversity and evenness and was a key driver of shifts in bacterial community structure. In samples containing oil or dispersant, sunlight greatly reduced abundance of the Cyanobacterium Synechococcus but increased the relative abundances of Alteromonas, Marinobacter, Labrenzia, Sandarakinotalea, Bartonella, and Halomonas. Dark samples with oil were represented by members of Thalassobius, Winogradskyella, Alcanivorax, Formosa, Pseudomonas, Eubacterium, Erythrobacter, Natronocella, and Coxiella. Both oil and Corexit inhibited the Candidatus Pelagibacter with or without sunlight exposure. For the first time, we demonstrated the effects of light in structuring microbial communities in water with oil and/or Corexit. Overall, our findings improve understanding of oil pollution in surface water, and provide unequivocal evidence that sunlight is a key factor in determining bacterial community composition and dynamics in oil polluted marine waters. C1 [Bacosa, Hernando P.; Liu, Zhanfei; Erdner, Deana L.] Univ Texas Austin, Inst Marine Sci, Port Aransas, TX 78373 USA. RP Erdner, DL (corresponding author), Univ Texas Austin, Inst Marine Sci, Port Aransas, TX 78373 USA. EM derdner@utexas.edu NR 0 TC 73 Z9 81 PD DEC 1 PY 2015 VL 6 AR 1326 DI 10.3389/fmicb.2015.01325 UT WOS:000366553800001 DA 2025-07-30 ER PT J AU Thiele, S Richter, M Balestra, C Glöckner, FO Casotti, R AF Thiele, Stefan Richter, Michael Balestra, Cecilia Gloeckner, Frank Oliver Casotti, Raffaella TI Taxonomic and functional diversity of a coastal planktonic bacterial community in a river-influenced marine area SO MARINE GENOMICS DT Article AB The Gulf of Naples is a dynamical area with intense exchanges between offshore oligotrophic and coastal eutrophic waters with frequent freshwater inputs. The Sarno River, one of the most polluted rivers in Europe, strongly contributes to the pollution of the area, discharging high amounts of heavy metals and organic wastes from heavily cultivated and industrial areas. This paper reports on the diversity and community structure of the marine residential Bacteria and Archaea of the Gulf of Naples in an area close to the river Sarno plume and investigates their small-scale taxonomic diversity and expression patterns as a proxy of potential metabolic activity using metagenomics and metatranscriptomics. Bacteria and Archaea were mainly represented by marine clades, with only minor contributors from freshwater ones. The community was dominated by Alpha- and Gammaproteobacteria, of which Rhodospirillales, Pelagibacteriales, and Oceanospirilalles were most represented. However, Alteromonadales and Rhodobacterales were the most active, despite their relative lower abundance, suggesting that they are important for overall ecosystem functioning and nutrient cycling. Nitrification and a reversed form of dissimilatory sulfate reduction were the major metabolic processes found in the metatrascriptomes and were mainly associated to NitrosopuMilaleS and Pelagibacter, respectively. No clear indication of transcripts related to stress induced by heavy metals or organic pollutants was found. In general, despite the high loads of pollutants discharged continuously by the Sarno River, the microbial community did not show marks of stress-induced changes neither structural nor functional, thus suggesting that this river has little or no effect on the planktonic bacterial community of the Gulf of Naples. (C) 2016 Elsevier B.V. All rights reserved. C1 [Thiele, Stefan; Balestra, Cecilia; Casotti, Raffaella] Stn Zool Anton Dohrn, I-80121 Naples, Italy. [Richter, Michael; Gloeckner, Frank Oliver] Max Planck Inst Marine Microbiol, Celsiusstr 1, D-28359 Bremen, Germany. [Gloeckner, Frank Oliver] Jacobs Univ Bremen gGmbH, Campus Ring 1, D-28759 Bremen, Germany. 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Ames, Tyler D. Breaker, Ronald R. TI A variant riboswitch aptamer class for S-adenosylmethionine common in marine bacteria SO RNA DT Article AB Riboswitches that sense S-adenosylmethionine (SAM) are widely distributed throughout a variety of bacterial lineages. Four classes of SAM-binding riboswitches have been reported to date, constituting the most diverse collection of riboswitch classes that sense the same compound. Three of these classes, termed SAM-I, SAM-II, and SAM-III represent unique structures that form distinct binding pockets for the ligand. SAM-IV riboswitches carry different conserved sequence and structural features compared to other SAM riboswitches, but nucleotides and substructures corresponding to the ligand binding pocket are identical to SAM-I aptamers. In this article, we describe a fifth class of SAM binding aptamer, which we have termed SAM-V. SAM-V was discovered by analyzing GC-rich intergenic regions preceding metabolic genes in the marine alpha-proteobacterium "Candidatus Pelagibacter ubique.'' Although the motif is nearly unrepresented in cultured bacteria whose genomes have been completely sequenced, SAM-V is prevalent in marine metagenomic sequences. The consensus sequence and structure of SAM-V show some similarities to that of the SAM-II riboswitch, and it is likely that the two aptamers form similar ligand binding pockets. In addition, we identified numerous examples of a tandem SAM-II/SAM-V aptamer architecture. In this arrangement, the SAM-II aptamer is always positioned 5' of the SAM-V aptamer and the SAM-II aptamer is followed by a predicted intrinsic transcription terminator stem. The SAM-V aptamer, however, appears to use a ribosome binding site occlusion mechanism for genetic regulation. 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Geraki, Kalotina Scanlan, David J. Zubkov, Mikhail, V TI Accumulation of ambient phosphate into the periplasm of marine bacteria is proton motive force dependent SO NATURE COMMUNICATIONS DT Article AB Bacteria acquire phosphate (P-i) by maintaining a periplasmic concentration below environmental levels. We recently described an extracellular P-i buffer which appears to counteract the gradient required for P-i diffusion. Here, we demonstrate that various treatments to outer membrane (OM) constituents do not affect the buffered P-i because bacteria accumulate P-i in the periplasm, from which it can be removed hypo-osmotically. The periplasmic P-i can be gradually imported into the cytoplasm by ATP-powered transport, however, the proton motive force (PMF) is not required to keep P-i in the periplasm. In contrast, the accumulation of P-i into the periplasm across the OM is PMF-dependent and can be enhanced by light energy. Because the conventional mechanism of P-i-specific transport cannot explain P-i accumulation in the periplasm we propose that periplasmic P-i anions pair with chemiosmotic cations of the PMF and millions of accumulated P-i pairs could influence the periplasmic osmolarity of marine bacteria. The ubiquitous oceanic bacteria harbour an external phosphate buffer for modulating phosphate (P-i) uptake. Here, using both oceanic SAR11, Prochlorococcus and Synechococcus strains as a model, the authors show that the P-i buffer accumulation in the periplasm is proton motive force-dependent and can be enhanced by light energy. C1 [Kamennaya, Nina A.; Zubkov, Mikhail, V] Natl Oceanog Ctr, Southampton SO14 3ZH, Hants, England. [Kamennaya, Nina A.; Scanlan, David J.] Univ Warwick, Sch Life Sci, Gibbet Hill, Coventry CV4 7AL, W Midlands, England. [Geraki, Kalotina] Harwell Sci & Innovat Campus, Diamond Light Source Ltd, Didcot OX11 0DE, Oxon, England. 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Wang, Kui Evans, Sarah E. Chen, Feng TI High temporal but low spatial heterogeneity of bacterioplankton in the Chesapeake bay SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Compared to freshwater and the open ocean, less is known about bacterioplankton community structure and spatiotemporal dynamics in estuaries, particularly those with long residence times. The Chesapeake Bay is the largest estuary in the United States, but despite its ecological and economic significance, little is known about its microbial community composition. A rapid screening approach, ITS (internal transcribed spacer)-LH (length beterogeneity)-PCR, was used to screen six rRNA operon (16S rRNA,-ITS-23S rRNA) clone libraries constructed from bacterioplankton collected in three distinct regions of the Chesapeake Bay over two seasons. 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Rubin-Blum, Maxim Rahav, Eyal Kiro, Yael TI Allochthonous Groundwater Microorganisms Affect Coastal Seawater Microbial Abundance, Activity and Diversity SO JOURNAL OF GEOPHYSICAL RESEARCH-BIOGEOSCIENCES DT Article AB Submarine groundwater discharge (SGD) is a globally important process supplying nutrients and trace elements to the coastal environment, thus playing a pivotal role in sustaining marine primary productivity. Along with nutrients, groundwater also contains allochthonous microbes that are discharged from the terrestrial subsurface into the sea. Currently, little is known about the interactions between groundwater-borne and coastal seawater microbial populations, and groundwater microbes' role upon introduction to coastal seawater populations. Here, we investigated seawater microbial abundance, activity and diversity in a site strongly influenced by SGD. In addition, through laboratory-controlled bottle incubations, we mimicked different mixing scenarios between groundwater and seawater. Our results demonstrate that the addition of 0.1 mu m filtered groundwater stimulated heterotrophic activity and increased microbial abundance compared to control coastal seawater, whereas 0.22 mu m filtration treatments induced primary productivity and Synechococcus growth. 16S rRNA gene sequencing showed a strong shift from a SAR11-rich community in the control samples to Rhodobacteraceae dominance in the <0.1 mu m treatment, in agreement with Rhodobacteraceae enrichment in the SGD field site. These results suggest that microbes delivered by SGD may affect the abundance, activity and diversity of intrinsic microbes in coastal seawater, highlighting the cryptic interplay between groundwater and seawater microbes in coastal environments, which has important implications for carbon cycling. 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R. Van Mooy, B. A. S. Mincer, T. J. TI Characterization of bacterial epibionts on the cyanobacterium Trichodesmium SO AQUATIC MICROBIAL ECOLOGY DT Article AB Interest in Trichodesmium, a nitrogen-fixing genus of Cyanobacteria, has been fueled by its prominent role in the marine nitrogen cycle. However, it is often overlooked that Trichodesmium occur in the ocean as colonies and are only one member of a complex microbial consortium. In the present study, we used 16S rRNA gene sequences to phylogenetically classify the communities associated with 2 morphological types of Trichodesmium colonies, tufts and puffs, which were collected at the Bermuda Atlantic Time Series (BATS) site in the Sargasso Sea. Both Trichodesmium morphotypes were most closely related to T. thiebautii at >= 99% identity. Non-cyanobacterial sequences from both types of colonies were dominated by Flavobacteria, Sphingobacteria, and Alphaproteobacteria. However, the epibiotic communities possessed significantly lower diversity than bacterioplankton; major seawater planktonic taxa, such as the SAR11 clade and Archaea, were conspicuously absent. Moreover, several epibiotic taxa appeared to be novel. Among the tuft or puff 16S rRNA clone libraries constructed in the present study, epibionts sharing common operational taxonomic units at the 97 percent sequence identity (PSI) threshold or higher were not observed, presenting the possibility that these 2 morphologies could select for different epibiotic communities. While our data are representative of a single sample point in time and space, these data suggest that Trichodesmium possess an epibiotic microbial community of relatively low diversity, distinct from that observed in bacterial picoplankton. C1 [Hmelo, L. R.; Van Mooy, B. A. S.; Mincer, T. J.] Woods Hole Oceanog Inst, Dept Marine Chem & Geochem, Woods Hole, MA 02543 USA. 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Gammaproteobacteria populations increased significantly during bloom declines and then decreased post-bloom. Divergences in the microbial community composition during different bloom periods were the result of changes in Candidatus, Pelagibacter, Alteromonas, Rhodobacteraceae, Vibrio and Pseudoalteromonas populations. Sediminimonas qiaohouensis was the first bacterium shown to be significantly negatively correlated with A. sanguinea concentration. This study indicated that bacteria may play an important role in A. sanguinea-bloom regulation and provides a deeper insight into bacterial community succession during and after an A. sanguinea-bloom. (C) 2012 Elsevier B.V. All rights reserved. C1 [Tian, Yun] Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen 361005, Peoples R China. Xiamen Univ, Key Lab, Minist Educ Coastal & Wetland Ecosyst, Sch Life Sci, Xiamen 361005, Peoples R China. 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TI The Black Queen Hypothesis: Evolution of Dependencies through Adaptive Gene Loss SO MBIO DT Article AB Reductive genomic evolution, driven by genetic drift, is common in endosymbiotic bacteria. Genome reduction is less common in free-living organisms, but it has occurred in the numerically dominant open-ocean bacterioplankton Prochlorococcus and "Candidatus Pelagibacter," and in these cases the reduction appears to be driven by natural selection rather than drift. Gene loss in free-living organisms may leave them dependent on cooccurring microbes for lost metabolic functions. We present the Black Queen Hypothesis (BQH), a novel theory of reductive evolution that explains how selection leads to such dependencies; its name refers to the queen of spades in the game Hearts, where the usual strategy is to avoid taking this card. Gene loss can provide a selective advantage by conserving an organism's limiting resources, provided the gene's function is dispensable. 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The interactions between viruses and their hosts is one of the hot spots of marine ecology. Besides their important role in various ecosystems, viruses, especially bacteriophages and their gene pool, are of enormous interest for the development of new gene products with high innovation value. Various studies have been conducted in diverse ecosystems to understand microbial diversity and phage-host interactions; however, the Black Sea, especially the Eastern coastal area, remains among the least studied ecosystems in this regard. This study was aimed at to fill this gap by analyzing microbial diversity and bacteriophage-host interactions in the waters of Eastern Black Sea using a metagenomic approach. To this end, prokaryotic and viral metagenomic DNA from two sampling sites, Poti and Gonio, were sequenced on the Illumina Miseq platform and taxonomic and functional profiles of the metagenomes were obtained using various bioinformatics tools. Our metagenomics analyses allowed us to identify the microbial communities, with Proteobacteria, Cyanobacteria, Actinibacteria, and Firmicutes found to be the most dominant bacterial phyla and Synechococcus and Candidatus Pelagibacter phages found to be the most dominant viral groups in the Black Sea. As minor groups, putative phages specific to human pathogens were identified in the metagenomes. We also characterized interactions between the phages and prokaryotic communities by determining clustered regularly interspaced short palindromic repeats (CRISPR), prophage-like sequences, and integrase/excisionase sequences in the metagenomes, along with identification of putative horizontally transferred genes in the viral contigs. In addition, in the viral contig sequences related to peptidoglycan lytic activity were identified as well. 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However, microbial biomarkers for the municipal effluent instead of the fecal species from raw sewage have not been proposed. Meanwhile, bacterial taxa for degrading large amounts of input organics have not been fully understood. In this study, raw effluent and serial water samples were collected from the coastal dispersal of two sewage treatment plants in Xiamen, China. Free-living (FL) and particle-associated (PA) bacterial communities were analyzed via high-throughput sequencing of 16S rRNA gene and quantitative PCR to measure bacterial abundance. The PA bacterial communities in our samples exhibited higher cell abundance, alpha diversity, and population dynamics than the FL bacterial communities, which supports greater environmental significance of the PA bacterial communities. Two non-fecal but typical genera in activated sludge, Zoogloea and Dechloromonas, exhibited decreased but readily detectable abundance along the effluent dispersal distance. Furthermore, the dominating microbial species near the outfalls were related to well-known marine indigenous taxa, such as SAR11 clade, OM60 clade, low-GC Actinobacteria, and unclassified Flavobacteriales, as well as the less understood taxa like Pseudohongiella and Microbacteriaceae. It is interesting that these taxa exhibited two types of correlation patterns with COD concentration. Our study suggested Zoogloea as a potential indicator of municipal effluents and also proposed potential utilizers of residual effluent COD in marine environments. C1 [Liu, SiGuang; Luo, YuanRong; Huang, LingFeng] Xiamen Univ, Fujian Prov Key Lab Coastal Ecol & Environm Studi, Xiamen 361005, Peoples R China. [Liu, SiGuang; Luo, YuanRong; Huang, LingFeng] Fujian Inst Oceanog, Xiamen 361005, Peoples R China. [Luo, YuanRong; Huang, LingFeng] Xiamen Univ, Key Lab, Coll Environm & Ecol, Minist Educ Coastal & Wetland Ecosyst, Xiamen 361005, Peoples R China. 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Sun, Shulei Biers, Erin J. Moran, Mary Ann TI Abundant and diverse bacteria involved in DMSP degradation in marine surface waters SO ENVIRONMENTAL MICROBIOLOGY DT Article AB An expanded analysis of oceanic metagenomic data indicates that the majority of prokaryotic cells in marine surface waters have the genetic capability to demethylate dimethylsulfoniopropionate (DMSP). The 1701 homologues of the DMSP demethylase gene, dmdA, identified in the (2007) Global Ocean Sampling (GOS) metagenome, are sufficient for 58% (+/- 9%) of sampled cells to participate in this critical step in the marine sulfur cycle. This remarkable frequency of DMSP-demethylating cells is in accordance with biogeochemical data indicating that marine phytoplankton direct up to 10% of fixed carbon to DMSP synthesis, and that most of this DMSP is subsequently degraded by bacteria via demethylation. The GOS metagenomic data also revealed a new cluster of dmdA sequences (designated Clade E) that implicates marine gammaproteobacteria in DMSP demethylation, along with previously recognized alphaproteobacterial groups Roseobacter and SAR11. Analyses of G+C content and gene order indicate that lateral gene transfer is likely responsible for the wide distribution of dmdA among diverse taxa, contributing to the homogenization of biogeochemical roles among heterotrophic marine bacterioplankton. Candidate genes for the competing bacterial degradation process that converts DMSP to the climate-active gas dimethylsulfide (DMS) (dddD and dddL) occur infrequently in the (2007) GOS metagenome, suggesting either that the key DMS-producing bacterial genes are yet to be identified or that DMS formation by free-living bacterioplankton is insignificant relative to their demethylation activity. C1 [Sun, Shulei; Biers, Erin J.; Moran, Mary Ann] Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. [Howard, Erinn C.] Univ Georgia, Dept Microbiol, Athens, GA 30602 USA. RP Moran, MA (corresponding author), Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. 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Sequences affiliated with the freshwater P. necessarius cluster are among the most frequently detected in studies on bacterial diversity in freshwater ecosystems. Despite this frequent detection with culture-independent techniques and the cosmopolitan occurrence of members affiliated with this cluster, no isolates have been reported thus far. The isolated strains have been obtained from lakes, ponds, and rivers in central Europe, the People's Republic of China, and East Africa by use of the filtration-acclimatization method. The 16S rRNA gene sequences of the isolates are 98.8 to 100% identical to reference sequences obtained by various authors by use of culture-independent methods. The isolates, aerobic heterotrophs, grew on a wide range of standard complex media and formed visible colonies on agar plates. Thus, the previous lack of isolates cannot be explained by a lack of appropriate media. 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[Shi, Rongjun; Li, Jiajun; Qi, Zhanhui; Zhang, Zhe; Liu, Huaxue; Huang, Honghui] Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Key Lab Open Sea Fishery Dev, Minist Agr, 231 West Xingang Rd, Guangzhou 510300, Guangdong, Peoples R China. [Shi, Rongjun] Chinese Acad Sci, South China Sea Inst Oceanol, Guangdong Prov Key Lab Appl Marine Biol, Guangzhou 510300, Guangdong, Peoples R China. RP Huang, HH (corresponding author), Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Guangdong Prov Key Lab Fishery Ecol & Environm, 231 West Xingang Rd, Guangzhou 510300, Guangdong, Peoples R China.; Huang, HH (corresponding author), Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Key Lab Open Sea Fishery Dev, Minist Agr, 231 West Xingang Rd, Guangzhou 510300, Guangdong, Peoples R China. 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Microbial communities associated with animals are known to be key elements in the development of their hosts. In marine environments, these communities are largely under the influence of the surrounding water. In aquaculture, understanding the interactions existing between the microbiotas of farmed species and their rearing environment could help establish precise bacterial management. Method. In light of these facts, we studied the active microbial communities associated with the eggs and the nauplii of the Pacific blue shrimp (Litopenaeus stylirostris) and their rearing water. All samples were collected in September 2018, November 2018 and February 2019. After RNA extractions, two distinct Illumina HiSeq sequencings were performed. Due to different sequencing depths and in order to compare samples, data were normalized using the Count Per Million method. Results. We found a core microbiota made of taxa related to Aestuariibacter, Al-teromonas, Vibrio, SAR11, HIMB11, AEGEAN 169 marine group and Candidatus En-dobugula associated with all the samples indicating that these bacterial communities could be transferred from the water to the animals. We also highlighted specific bacterial taxa in the eggs and the nauplii affiliated to Pseudomonas, Corynebacterium, Acinetobacter, Labrenzia, Rothia, Thalassolituus, Marinobacter, Aureispira, Oleiphilus, Profundimonas and Marinobacterium genera suggesting a possible prokaryotic vertical transmission from the breeders to their offspring. This study is the first to focus on the active microbiota associated with early developmental stages of a farmed shrimp species and could serve as a basis to comprehend the microbial interactions involved throughout the whole rearing process. 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Huifang Ma, Xu Wang, Haibo Shi, Baoyou TI Metabolic response of bacterial community to sodium hypochlorite and ammonia nitrogen affected the antibiotic resistance genes in pipelines biofilm SO WATER RESEARCH DT Article AB The biofilm is important for the antibiotic resistance genes (ARGs) propagation in drinking water pipelines. This study investigated the influence of chlorine disinfection and ammonia nitrogen on the ARGs in pipelines biofilm using metagenomic and metabolomics analysis. Chlorine disinfection reduced the relative abundance of unclassified_c_Actinobacteria, Acidimicrobium, and Candidatus_Pelagibacter to 394-430 TPM, 114-123 TPM, and 4954 TPM, respectively. Correspondingly, the ARGs Saur_rpoC_DAP, macB, and mfd was reduced to 8-12 TPM, 81-92 TPM and 30-35 TPM, respectively. The results of metabolomics suggested that chlorine disinfection suppressed the pathways of ABC transporters, fatty acid biosynthesis, biosynthesis of unsaturated fatty acids, and biosynthesis of amino acids. These pathways were related to the cell membrane integrality and extracellular polymeric substances (EPS) secretion. Chlorine disinfection induced the decrease of EPS-related genes, resulting in the lower relative abundance of bacterial community and their antibiotic resistance. However, added approximately 0.5 mg/L NH3-N induced up-regulation of these metabolic pathways. In addition, NH3-N addition increased the relative abundance of enzymes related to inorganic and organic nitrogen metabolic pathway significantly, such as ammonia monooxygenase, glutamine synthetase, and glutamate synthase. Due to the EPS protection and nitrogen metabolism, the relative abundance of the main bacterial genera and the related ARGs increased to the level equal to that in pipelines biofilm with no disinfection. Therefore, NH3-N reduced the ARGs removal efficiency of chlorine disinfection. It is necessary to take measures to improve the removal rate of NH3-N and ARGs for preventing their risks in drinking water. C1 [Wang, Min; Ma, Xu; Wang, Haibo; Shi, Baoyou] Chinese Acad Sci, Res Ctr Ecoenvironm Sci, Key Lab Drinking Water Sci & Technol, Beijing 100085, Peoples R China. [Sun, Huifang] Shanxi Univ, Inst Resources & Environm Engn, Taiyuan 030006, Shanxi, Peoples R China. 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At a temperate time-series site in the Tasman Sea, we observed significant reductions in standing stocks of total carbon and chlorophylla, and a shift towards smaller phytoplankton and carnivorous copepods, associated with the seasonal impact of the EAC microbial assemblage. In light of the substantial shifts in microbial assemblage structure and function associated with the EAC, we conclude that climate-driven expansions of WBCs will expand the range of tropical oligotrophic microbes, and potentially profoundly impact the trophic status of temperate waters. C1 [Messer, Lauren F.] Univ Queensland, Australian Ctr Ecogen, Sch Chem & Mol Biosci, Brisbane, Qld, Australia. [Ostrowski, Martin; Paulsen, Ian] Univ Technol Sydney, Climate Change Cluster, Sydney, NSW, Australia. [Ostrowski, Martin; Doblin, Martina A.; Seymour, Justin R.] Macquarie Univ, Dept Mol Sci, Sydney, NSW, Australia. [Petrou, Katherina] Univ Technol Sydney, Sch Life Sci, Sydney, NSW, Australia. 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Mar Martin-Cardona, Celia Lalucat, Jorge Bosch, Rafael TI Bacterial diversity, composition and dynamics in and around recreational coastal areas SO ENVIRONMENTAL MICROBIOLOGY DT Article AB A comparative study on the composition of bacterial communities in a coastal area in the West Mediterranean receiving the impact of recreation-derived activities (from a marina and a beach) was performed by terminal-restriction fragment length polymorphism (T-RFLP) of 16S rDNA along spatial and temporal scales. Interpolation of concentration of hydrophobic compounds, chlorophyll and bacterial cells in seawater over the geography of the sampling area using geographic information systems techniques (GIS) allowed the delineation of two different habitats: bay and marina (with low and high levels of impact respectively), and a transition zone between them. Accordingly, the 16S rDNA T-RFLP profiles of bacterial communities in the area differed mainly spatially, with gradual changes in community composition and structure when approaching the beach and marina. Bacterial communities in impacted areas had higher diversity and equitability, as well as different composition. The main bacterial populations inferred in bay samples, which were members of the Alphaproteobacteria (mainly SAR11 and Roseobacter groups), were replaced by a different population of the Roseobacter clade, and members of the Gammaproteobacteria and Bacteroidetes in more impacted areas. There were also differences in the dynamics of bacterial communities. 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Marine Roseobacter group is an abundant heterotrophic bacterial group that is involved in many major element cycles, especially carbon and sulfur. Members of the Roseobacter CHUG (Clade Hidden and Underappreciated Globally) lineage are globally distributed and are activated in pelagic marine environments. In this study, we isolated and characterized a phage, CRP-810, that infects the CHUG strain FZCC0198. The genome of CRP-810 was dissimilar to those of other known phages. Additionally, 251 uncultured viral genomes (UViGs) closely related to CRP-810 were obtained from the uncultivated marine viral contig databases. Comparative genomic and phylogenetic analyses revealed that CRP-810 and these related UViGs exhibited conserved genome synteny, representing a new phage family with at least eight subgroups. Most of the CRP-810-type phages contain an integrase gene, and CRP-810 can be integrated into the host genome. Further analysis revealed that three CRP-810-type members were prophages found in the genomes of marine SAR11, Poseidonocella, and Sphingomonadaceae. Finally, viromic read-mapping analysis showed that CRP-810-type phages were globally distributed and displayed distinct biogeographic patterns related to temperature and latitude. Many members with a lower G + C content were mainly distributed in the trade station, whereas members with a higher G + C content were mainly distributed in polar and westerlies station, indicating that the niche differentiation of phages was subject to host adaptation. Collectively, these findings identify a novel phage family and expand our understanding of phylogenetic diversity, evolution, and biogeography of marine phages. C1 [Wu, Zuqing; Guo, Luyuan; Wu, Ying; Yang, Mingyu; Du, Sen; Shao, Jiabing; Zhang, Zefeng; Zhao, Yanlin] Fujian Agr & Forestry Univ, Coll JunCao Sci & Ecol, Fujian Prov Key Lab Agroecol Proc & Safety Monitor, Fuzhou, Peoples R China. 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The percentage of bacteria assimilating DMSP-S showed a strong seasonal pattern, with a steady increase from winter (8 +/- 5%) to summer (23 +/- 3%). The same seasonal pattern was observed for the rate of DMSP-S assimilation. The annual average percentage of DMSP-S-assimilating bacteria (16 +/- 8%) was lower than the corresponding percentage of leucine-assimilating cells (35 +/- 16%), suggesting that not all bacteria synthesizing protein incorporated DMSP-S. Smaller differences between both percentages were recorded in summer. Members of the Alphaproteobacteria (Roseobacter and SAR11) and Gammaproteobacteria groups accounted for most of bacterial DMSP-S-assimilating cells over the year. All major bacterial groups showed an increase of the percentage of cells assimilating DMSP-S during summer, and contributed to the increase of the DMSP-S assimilation rate in this period. In these primarily P-limited waters, enrichment with P + DMSP resulted in a stimulation of bacterial heterotrophic production comparable to, or higher than, that with P + glucose in summer, while during the rest of the year P + glucose induced a stronger response. This suggested that DMSP was more important a S and C source for bacteria in the warm stratified season. Overall, our results suggest that DMSP-S assimilation is controlled by the contribution of DMSP to S (and C) sources rather than by the phylogenetic composition of the bacterioplankton. C1 CSIC, Inst Ciencies Mar, E-08003 Barcelona, Catalonia, Spain. Univ Kalmar, Dept Biol & Environm Sci, SE-39182 Kalmar, Sweden. Max Planck Inst Marine Microbiol, D-28359 Bremen, Germany. RP Simó, R (corresponding author), CSIC, Inst Ciencies Mar, Pg Maritim Barceloneta 37-49, E-08003 Barcelona, Catalonia, Spain. 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High-throughput DNA sequencing of full-length 16S rRNA was employed to construct a genomic database. The results of the bioinformatic analysis revealed that two stations, 48 and 50 (Makassar Strait), exhibited a more similar community structure of deep-sea bacteria than did station 33 (Lombok Strait). Among the predominant phyla found at a depth of 1000 m, the top ten were Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Planctomycetes, Acidobacteria, Nitrospinae, Verrucomicrobia, Candidatus Melainabacteria, and Cyanobacteria. Furthermore, the genera Colwellia, Moritella, Candidatus Pelagibacter, Alteromonas, and Psychrobacter consistently appeared at all three stations, albeit with varying relative abundance values. These bacterial genera share common characteristics, such as psychrophilic, halophilic, and piezophilic tendencies, and are commonly found in deep-sea ecosystem. The environmental conditions at a depth of 1000 m were relatively stable, with an average pressure 10 MPa, temperature 4.68 degrees C, salinity 34.58 PSU, pH 8.06, chlorophyll-a 0.29 mu g/L, nitrate 3.19 mu mol/L, phosphate 6.32 mu mol/L and dissolved oxygen (DO) 2.90 mg/L. The bacterial community structures at the three sampling stations located at the same depth (1000 m) exhibited similarities, as indicated by the closely aligned similarity index values. C1 [Siallagan, Zen Ladestam; Nugrahapraja, Husna; Dwivany, Fenny M.] Inst Teknol Bandung, Sch Life Sci & Technol, Doctoral Program Biol, Jl Ganesha 10, Bandung 40132, Indonesia. [Siallagan, Zen Ladestam; Fadli, Muhammad; de Fretes, Charlie Ester; Opier, Rafidha Dh Ahmad; Radjasa, Ocky Karna] Natl Res & Innovat Agcy, Res Ctr Deep Sea, Jakarta 14430, Indonesia. [Suhardi, V. Sri Harjati; Nugrahapraja, Husna; Radjasa, Ocky Karna; Dwivany, Fenny M.] Inst Teknol Bandung, Ctr Biosci & Biotechnol, Jl Ganesha 10, Bandung 40132, Indonesia. [Susanto, R. Dwi] Univ Maryland, Dept Atmospher & Ocean Sci, College Pk, MD 20742 USA. [Wei, Zexun] Minist Nat Resources, Inst Oceanog 1, Qingdao, Peoples R China. [Wei, Zexun] Minist Nat Resources, Key Lab Marine Sci & Numer Modeling, Qingdao, Peoples R China. [Fadli, Muhammad] Univ Pattimura, Ctr Collaborat Res Aquat Ecosyst Eastern Indonesia, Ambon 97233, Indonesia. RP Siallagan, ZL; Dwivany, FM (corresponding author), Inst Teknol Bandung, Sch Life Sci & Technol, Doctoral Program Biol, Jl Ganesha 10, Bandung 40132, Indonesia.; Siallagan, ZL; Radjasa, OK (corresponding author), Natl Res & Innovat Agcy, Res Ctr Deep Sea, Jakarta 14430, Indonesia.; Radjasa, OK; Dwivany, FM (corresponding author), Inst Teknol Bandung, Ctr Biosci & Biotechnol, Jl Ganesha 10, Bandung 40132, Indonesia. 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However, the effect of geographic distance and its related factors (i.e., salinity and nutrients) on bacterial diversity, co-occurrence, as well as community assembly on the abundant and rare species scale remained to be discussed. In this study, we deci-phered the bacterial dynamics and ecological processes in the anthropogenic estuary of the Haihe River, in China. The results revealed that salinity was the major abiotic factor to shape bacterial community composition, diversity, and network complexity, and the role of biotic cytometric characteristics was considerable as well. Furthermore, the salinity gradient tended to select bacteria with low nucleic acid (LNA) such as Candidatus Pelagibacter and Longivirga, and abundant bacteria were more sensitive to the salinity gradient in comparison with rare ones. The similar keystone number and network topology values indicated that both abundant and rare bacteria played important ecological functions in degrading pollutants, developing co-occurrence networks, and maintaining compositional stability and complexity. In addition, drift and dispersion limitation dominated the ecological processes in the community assembly, suggesting that although salinity caused the migration of species, its ecological role was still weaker than that of stochastic processes. Further, niche differentiation contributed to different stochastic components of abundant and rare species. In high-salinity areas, rare bacteria with small population sizes but high diversity were enriched, which made the bacterial community more affected by ecological drift. Our study illustrated the different responses of abundant and rare bacteria to salinity and the assembly process of bacterial communities in anthropogenic estuary, deepening the understanding of their ecological processes in the estuarine ecosystem. C1 [Shi, Ziyue; Liu, Jie] Lanzhou Univ, Coll Pastoral Agr Sci & Technol, Collaborat Innovat Ctr Western Ecol Safety, Ctr Grassland Microbiome,State Key Lab Herbage Imp, Lanzhou 730000, Peoples R China. [Wang, Yingying; Liu, Jie] Nankai Univ, Coll Environm Sci & Engn, Key Lab Pollut Proc & Environm Criteria, Minist Educ, Tianjin 300071, Peoples R China. [Ma, Lili] Southwest Petr Univ, Coll Chem & Chem Engn, Chengdu 610500, Sichuan, Peoples R China. [Wang, Yingying] Nankai Univ, Coll Environm Sci & Engn, 94 Weijin Rd, Tianjin 300071, Peoples R China. [Liu, Jie] 222 TianShui Rd South, Lanzhou, Gansu, Peoples R China. 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We report here a shotgun metagenomic analysis of the year-round variation in free-living bacterioplankton collected across the entire length of the bay. Phylogenetic differences among spring, summer, autumn and winter bacterioplankton suggested that members of Proteobacteria tended to decrease at high water temperatures and increase at low temperatures. It was revealed that Candidatus Pelagibacter varied seasonally, reaching as much as 60% of all sequences at the genus level in the surface waters during winter. This increase was more evident in the deeper waters, where they reached up to 75%. The relative abundance of Planktomarina also rose during winter and fell during summer. A significant component of the winter bacterioplankton community was Archaea (mainly represented by Nitrosopumilus), as their relative abundance was very low during spring and summer but high during winter. In contrast, Actinobacteria and Cyanobacteria appeared to be higher in abundance during high-temperature periods. It was also revealed that Bacteroidetes constituted a significant component of the summer bacterioplankton community, being the second largest bacterial phylum detected in the Ofunato Bay. Its members, notably Polaribacter and Flavobacterium, were found to be high in abundance during spring and summer, particularly in the surface waters. Principal component analysis and hierarchal clustering analyses showed that the bacterial communities in the Ofunato Bay changed seasonally, likely caused by the levels of organic matter, which would be deeply mixed with surface runoff in the winter. C1 [Reza, Md Shaheed; Kobiyama, Atsushi; Yamada, Yuichiro; Ikeda, Yuri; Ikeda, Daisuke; Mizusawa, Nanami; Ikeo, Kazuho; Sato, Shigeru; Ogata, Takehiko; Jimbo, Mitsuru; Kudo, Toshiaki; Watabe, Shugo] Kitasato Univ, Sch Marine Biosci, Minami Ku, Sagamihara, Kanagawa 2520373, Japan. [Kaga, Shinnosuke; Watanabe, Shiho; Naiki, Kimiaki; Kaga, Yoshimasa] Iwate Fisheries Technol Ctr, Kamaishi, Iwate 0260001, Japan. [Mineta, Katsuhiko; Bajic, Vladimir; Gojobori, Takashi] King Abdullah Univ Sci & Technol, Computat Biosci Res Ctr, Thuwal 239556900, Saudi Arabia. [Reza, Md Shaheed] Bangladesh Agr Univ, Dept Fisheries Technol, Mymensingh 2202, Bangladesh. [Ikeo, Kazuho] Natl Inst Genet, Mishima, Shizuoka 4118540, Japan. [Kaga, Shinnosuke] Iwate Prefectural Govt, Ofunato Fisheries Promot Ctr, Ofunato, Iwate 0228502, Japan. RP Gojobori, T (corresponding author), King Abdullah Univ Sci & Technol, Computat Biosci Res Ctr, Thuwal 239556900, Saudi Arabia.; Gojobori, T (corresponding author), King Abdullah Univ Sci & Technol, Biol & Environm Sci & Engn, Thuwal 239556900, Saudi Arabia.; Watabe, S (corresponding author), Kitasato Univ, Dept Marine Biochem, Sch Marine Biosci, Minami Ku, Sagamihara, Kanagawa 2520373, Japan. 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Hammar, Katherine M. Ducklow, Hugh W. Yager, Patricia L. Post, Anton F. TI Phaeocystis antarctica blooms strongly influence bacterial community structures in the Amundsen Sea polynya SO FRONTIERS IN MICROBIOLOGY DT Article AB Rising temperatures and changing winds drive the expansion of the highly productive polynyas (open water areas surrounded by sea ice) abutting the Antarctic continent. Phytoplankton blooms in polynyas are often dominated by the haptophyte Phaeocystis antarctica, and they generate the organic carbon that enters the resident microbial food web. Yet, little is known about how Phaeocystis blooms shape bacterial community structures and carbon fluxes in these systems. We identified the bacterial communities that accompanied a Phaeocystis bloom in the Amundsen Sea polynya during the austral summers of 20072008 and 20102011. These communities are distinct from those determined for the Antarctic Circumpolar Current (ACC) and off the Palmer Peninsula. 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Here, we explored viral life strategy and its potential effect on the bacterial community by experimental incubations of eutrophic coastal waters under lysogen-induced and non-induced treatments. The lysogen-induced treatment showed relatively constant viral and bacterial abundances, lytic and lysogenic viral production throughout the experimental period, together with the progressive declines in not only the relative abundances for SAR11, Rhodobacteraceae, Alteromonadaceae, and SAR86 but the bacterial community diversity. Conversely, the non-induced treatment observed the marked variation in the abundances of viruses, bacteria and cells with high nucleic acid content over the time course of incubation, which was congruent with the drastic shift in lytic and lysogenic viral production as well as the succession of bacterial community. 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Salcher, Michaele M. Pernthaler, Jakob TI Fluorescence in situ hybridization and sequential catalyzed reporter deposition (2C-FISH) for the flow cytometric sorting of freshwater ultramicrobacteria SO FRONTIERS IN MICROBIOLOGY DT Article AB Flow cytometric sorting is a powerful tool to physically separate cells within mixed microbial communities. If combined with phylogenetic staining (fluorescence in situ hybridization, FISH) it allows to specifically sort defined genotypic microbial populations from complex natural samples. However, the targeted enrichment of freshwater ultramicrobacteria, such as members of the LD12 clade of Alphaproteobacteria (SAR11-IIIb), is still challenging. Current FISH protocols, even in combination with signal amplification by catalyzed reporter deposition (CARD), are not sufficiently sensitive for the distinction of these bacteria from background noise by flow cytometry, presumably due to their low ribosome content and small cell sizes. We, therefore, modified a CARD based flow sorting protocol with the aim of increasing its sensitivity to a level sufficient for ultramicrobacteria. This was achieved by a second signal amplification step mediated by horseradish peroxidase labeled antibodies targeted to the fluorophores that were previously deposited by CARD-FISH staining. The protocol was tested on samples from an oligo-mesotrophic lake. Ultramicrobacteria affiliated with LD12 Alphaproteobacteria could be successfully sorted to high purity by flow cytometry. The ratios of median fluorescence signal to background ranged around 20, and hybridization rates determined by flow cytometry were comparable to those obtained by fluorescence microscopy. Potential downstream applications of our modified cell staining approach range from the analysis of microdiversity within 16S rRNA-defined populations to that of functional properties, such as the taxon-specific incorporation rates of organic substrates. 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Logares, Ramiro Deutschmann, Ina Maria Scharek, Renate Moran, Xose Anxelu G. Gonzalez, Felipe Alonso-Saez, Laura TI Novel Interactions Between Phytoplankton and Bacteria Shape Microbial Seasonal Dynamics in Coastal Ocean Waters SO FRONTIERS IN MARINE SCIENCE DT Article AB Trophic interactions between marine phytoplankton and heterotrophic bacteria are at the base of the biogeochemical carbon cycling in the ocean. However, the specific interactions taking place between phytoplankton and bacterial taxa remain largely unexplored, particularly out of phytoplankton blooming events. Here, we applied network analysis to a 3.5-year time-series dataset to assess the specific associations between different phytoplankton and bacterial taxa along the seasonal scale, distinguishing between free-living and particle-attached bacteria. 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Lefort, Thomas Hofer, Julia Sommaruga, Ruben TI Effect of natural sunlight on bacterial activity and differential sensitivity of natural bacterioplankton groups in northwestern Mediterranean coastal waters SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB We studied the effects of natural sunlight on heterotrophic marine bacterioplankton in short-term experiments. We used a single-cell level approach involving How cytometry combined with physiological probes and microautoradiography to determine sunlight effects on the activity and integrity of the cells. After 4 h of sunlight exposure, most bacterial cells maintained membrane integrity and viability as assessed by the simultaneous staining with propidium iodide and SYBR green I. In contrast, a significant inhibition of heterotrophic bacterial activity was detected, measured by 5-cyano-2,3 ditolyl tetrazolium chloride reduction and leucine incorporation. We applied microautoradiography combined with catalyzed reporter deposition-fluorescence in situ hybridization to test the sensitivity of the different bacterial groups naturally occurring in the Northwestern Mediterranean to sunlight. Members of the Gammaproteobacteria and Bacteroidetes groups appeared to be highly resistant to solar radiation, with small changes in activity after exposure. On the contrary, Alphaproteobacteria bacteria were more sensitive to radiation as measured by the cell-specific incorporation of labeled amino acids, leucine, and ATE Within Alphaproteobacteria, bacteria belonging to the Roseobacter group showed higher resistance than members of the SAR11 cluster. The activity of Roseobacter was stimulated by exposure to photosynthetic available radiation compared to the dark treatment. 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Montgomery, Andrew Montoya, Joseph P. Joye, Samantha B. TI Significance of Acetate as a Microbial Carbon and Energy Source in the Water Column of Gulf of Mexico: Implications for Marine Carbon Cycling SO GLOBAL BIOGEOCHEMICAL CYCLES DT Article AB Acetate is a key intermediate of organic matter mineralization, but its metabolism remains largely unconstrained in the pelagic ocean. We conducted an integrated biogeochemical study to investigate microbial acetate cycling in the northern Gulf of Mexico with the goal of elucidating the importance of acetate as a carbon and energy source. Acetate was used primarily as an energy source, as evidenced by observed oxidation rates (rate constant k: 0.06-0.22 day(-1)) that varied between 42% and 96% of total biological acetate uptake (i.e., assimilation + oxidation; k: 0.06-0.34 day(-1)). 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Obernosterer, Ingrid Zemb, Olivier Lebaron, Philippe TI Major differences of bacterial diversity and activity inside and outside of a natural iron-fertilized phytoplankton bloom in the Southern Ocean SO ENVIRONMENTAL MICROBIOLOGY DT Article AB One of the first comparisons of a natural iron fertilized bloom with a high-nutrient low-chlorophyll (HNLC) site was undertaken during the Kerguelen ocean and plateau compared study (KEOPS) cruise. To understand better the bacteria-phytoplankton relationship in the context of natural iron fertilization, bacterial diversity and activity was investigated in the bloom and in the adjacent HNLC region by 16S rDNA clone libraries and by single strand conformation polymorphism (SSCP) analysis. Both libraries were dominated by Alphaproteobacteria, Gammaproteobacteria and the Cytophaga-Flavobacteria-Bacteroides group. Cluster analysis at 99% sequence similarity yielded several microdiverse clusters and revealed striking differences between the two libraries. In the bloom, the dominant operational taxonomic units (OTUs) were the Roseobacter NAC11-7 cluster, SAR92 and a Cytophaga-Flavobacteria-Bacteroides cluster related to the agg58 group, whereas in the HNLC region, SAR11, Roseobacter RCA and Polaribacter dominated. SSCP analysis of 16S rDNA and 16S rRNA revealed contrasting dynamics of three different Roseobacter OTUs. Roseobacter NAC11-7 and NAC11-6 had higher relative abundances and activities in the bloom compared with the HNLC site and NAC11-6 was only detected at the decline of the bloom concomitant with a shift in phytoplankton composi tion. In contrast, Roseobacter RCA was relatively abundant and active both inside and outside of the bloom. 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The abundance of alpha- and beta-proteobacteria and Actinobacteria varied systematically in the estuary and mirrored the pattern seen in lakes and oceans. Beta-proteobacteria and Actinobacteria were abundant in the Delaware River but were less so in the marine waters of the Delaware Bay. In contrast, alpha-proteobacteria, including the SAR11 clade, were most abundant in the Bay and rare in the Delaware River. Actinobacteria were active in assimilating thymidine and leucine and appeared to contribute substantially to bacterial production in the Delaware River. Among the several biogeochemical parameters we examined, only salinity accounted for a substantial portion of the variation in abundance of these bacterial groups. However, relative abundance of these groups often varied independently of salinity. Cytophaga-like bacteria were often abundant throughout the estuary, but they did not vary systematically over the estuarine gradient, unlike the other dominant bacterial groups. 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Here we examine population-level heterogeneity within abundant and ubiquitous freshwater bacterial groups such as the acI Actinobacteria and LD12 Alphaproteobacteria (the freshwater sister clade to the marine SAR11) using 33 single-cell genomes and a 5-year metagenomic time series. The single-cell genomes grouped into 15 monophyletic clusters (termed "tribes") that share at least 97.9% 16S rRNA identity. Distinct populations were identified within most tribes based on the patterns of metagenomic read recruitments to single-cell genomes representing these tribes. Genetically distinct populations within tribes of the acI Actinobacterial lineage living in the same lake had different seasonal abundance patterns, suggesting these populations were also ecologically distinct. In contrast, sympatric LD12 populations were less genetically differentiated. 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Plume communities were highly similar to each other with significant differences in community membership only between Kilo Moana and Mariner, two vents that are separated by extremes in depth, latitude and geochemistry. Systematic sampling of waters surrounding the vents revealed that species richness and phylogenetic diversity was typically highest near the vent orifice, implying mixing of microbial communities from the surrounding habitats. Above-plume background communities were primarily dominated by SAR11, SAR324 and MG-I Archaea, while SUP05, Sulfurovum, Sulfurimonas, SAR324 and Alteromonas were abundant in plume and near-bottom background communities. These results show that the ubiquitous water-column microorganisms populate plume communities, and that the composition of background seawater exerts primary influence on plume community composition, with secondary influence from geochemical and/or physical properties of vents. 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Kan, Jinjun Inskeep, William P. Lovalvo, Dave Varley, John Mathur, Eric Nealson, Kenneth Gorby, Yuri Jiang, Hongchen LaFracois, Toben McDermott, Timothy R. TI Yellowstone Lake: high-energy geochemistry and rich bacterial diversity SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Yellowstone Lake is central to the balanced functioning of the Yellowstone ecosystem, yet little is known about the microbial component of its food chain. A remotely operated vehicle provided video documentation (http://www.tbi.montana.edu/media/videos/) and allowed sampling of dilute surface zone waters and enriched lake floor hydrothermal vent fluids. Vent emissions contained substantial H2S, CH4, CO2 and H-2, although CH4 and H-2 levels were also significant throughout the lake. Pyrosequencing and near full-length sequencing of Bacteria 16S rRNA gene diversity associated with two vents and two surface water environments demonstrated that this lake contains significant bacterial diversity. Biomass was size-fractionated by sequentially filtering through 20-mu m-, 3.0-mu m-, 0.8-mu m- and 0.1-mu m-pore-size filters, with the > 0.1 to < 0.8 mu m size class being the focus of this study. Major phyla included Acidobacteria, Actinobacteria, Bacteroidetes, alpha- and beta-Proteobacteria and Cyanobacteria, with 21 other phyla represented at varying levels. Surface waters were dominated by two phylotypes: the Actinobacteria freshwater acI group and an alpha-Proteobacteria clade tightly linked with freshwater SAR11-like organisms. We also obtained evidence of novel thermophiles and recovered Prochlorococcus phylotypes (97-100% identity) in one near surface photic zone region of the lake. The combined geochemical and microbial analyses suggest that the foundation of this lake's food chain is not simple. 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Jhirad, Nicholas Dodge, Jonathon Carlson, Craig A. Giovannoni, Stephen J. TI Marine bacterioplankton consortia follow deterministic, non-neutral community assembly rules SO AQUATIC MICROBIAL ECOLOGY DT Article AB The Bermuda Atlantic Time-series Study site provided an opportunity to study bacterial community assembly processes at 2 different depths, the surface and 200 m, in the upper mesopelagic, just below the euphotic zone. Over 100 monthly bacterioplankton DNA samples, from each depth, were analyzed using 16S rRNA gene sequences parsed with the custom software package PhyloAssigner. Co-occurrence networks, filtered for potential autocorrelation artifacts, were constructed for each depth. Network characteristics for the 2 depths were remarkably similar, and network parameters, such as connectance, were in the same range as previously published for ecological networks. Spectral clustering applied to similarity matrices based on exact connections revealed clusters of nodal taxonomic units (NTUs) that peaked at similar times, supporting deterministic, niche-based assembly. An algorithm that used hierarchical Dirichlet processes (HDPs) to model neutral communities based on learned parameters indicated that community assembly processes fit niche-based models at the metacommunity level for both depths. However, HDP analyses restricted to SAR11, SAR86, or SAR202 NTUs supported the neutral assembly hypothesis, suggesting that neutral process models may apply within some phylogenetic domains. To understand whether phylogenetically related taxa can substitute for one another in networks, we created a new metric, phylogenetically weighted connectivity, which considered the similarity of connections among near phylogenetic neighbors. This analysis suggested that phylogenetically similar lineages share similar network connections. 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Microbes participate in the energy exchange in coral ecosystems, which may affect coral resistance and resilience. However, there is still a lack of understanding regarding the microbial structure between corals and seawater. In this study, microbial structure and interactions in coral and seawater were studied using quantitative PCR and high- throughput sequencing. We found that the abundance and diversity of microbes in corals were higher than those in seawater. Corals, as nonfluidic ecosystems, limited the dispersal of microbes, causing broader stochasticity in the ecological shaping process, whereas ecological drift and homogeneous selection were the most important assembly mechanisms in seawater. The microbial niche width was larger in the coral group than in the seawater group, indicating strong adaptability. A group of microbes (e.g., Caldilineaceae, , Burkholderiaceae, , and Nitrosopumilaceae) ) may become microbial indicators which differentiate seawater samples (e.g., SAR11 and SAR86 clades, Cryomorphaceae), ), which may be due to separated habitats and nutrition. Coral microbes may participate in nitrogen-metabolism to maintain a nitrogen limited microenvironment. Nodes linked in the coral co- occurrence network showed coexisting interactions and higher complexity based on the comparison of topological values. Collectively, significant differences in microbial fractions were demonstrated observed in terms of diversity, composition, co-occurrence patterns, assembly processes and predicted functions between coral and seawater samples, showing potential microbial interactions and providing in-depth insights into the meta- community diversity in coral reef ecosystems. C1 [Zhao, Wenbin; Chen, Leiran; Huang, Xiaoyun; Liu, Jiwen; Zhang, Xiao-Hua; Wang, Xiaolei] Ocean Univ China, Coll Marine Life Sci & Frontiers Sci, Ctr Deep Ocean Multispheres & Earth Syst, Qingdao 266003, Peoples R China. [Niu, Wentao; Xiao, Jiaguang] Minist Nat Resources, Inst Oceanog 3, Lab Marine Biodivers Res, 178 Daxue Rd, Xiamen 361005, Peoples R China. [Zhao, Wenbin; Chen, Leiran; Huang, Xiaoyun; Liu, Jiwen; Zhang, Xiao-Hua; Wang, Xiaolei] Ocean Univ China, Inst Evolut & Marine Biodivers, Qingdao 266100, Peoples R China. [Niu, Wentao; Xiao, Jiaguang] Nansha Isl Coral Reef Ecosyst Natl Observat & Res, Guangzhou 510000, Peoples R China. [Zhao, Wenbin] Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao, Peoples R China. [Thompson, Fabiano] Fed Univ Rio de Janeiro UFRJ, Inst Biol, Rio De Janeiro, Brazil. [Thompson, Fabiano] Fed Univ Rio de Janeiro UFRJ, COPPE, Rio De Janeiro, Brazil. RP Wang, XL (corresponding author), Ocean Univ China, Coll Marine Life Sci & Frontiers Sci, Ctr Deep Ocean Multispheres & Earth Syst, Qingdao 266003, Peoples R China.; Xiao, JG (corresponding author), Minist Nat Resources, Inst Oceanog 3, Lab Marine Biodivers Res, 178 Daxue Rd, Xiamen 361005, Peoples R China.; Wang, XL (corresponding author), Ocean Univ China, Inst Evolut & Marine Biodivers, Qingdao 266100, Peoples R China.; Xiao, JG (corresponding author), Nansha Isl Coral Reef Ecosyst Natl Observat & Res, Guangzhou 510000, Peoples R China. 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De Freitas, Mayanne A. M. Lima, Michele S. Soares, Ana Carolina Leal, Camille Busch, Mileane De S. Tschoeke, Diogo A. Vidal, Luciana De O. Atella, Georgia C. Kruger, Ricardo H. Setubal, Joao Vasconcelos, Agnaldo A. De Mahiques, Michel M. Siegle, Eduardo Asp, Nils Edvin Cosenza, Carlos Hajdu, Eduardo De Rezende, Carlos E. Thompson, Cristiane C. Thompson, Fabiano L. TI Fluxes of the Amazon River plume nutrients and microbes into marine sponges SO SCIENCE OF THE TOTAL ENVIRONMENT DT Article AB Sponges have co-evolved with microbes for over 400 myr. Previous studies have demonstrated that sponges can be classified according to the abundance of microbes in their tissues as Low Microbial Abundance (LMA) and High Microbial Abundance (HMA). While LMA sponges rely mainly on water column microbes, HMA appear to rely much more on symbiotic fermentative and autotrophic microbes maintained in their tissues. However, it is unclear if this pattern holds when comparing different species of tropical sponges under extreme nutrient conditions and sediment loads in the water column, such as the Great Amazon Reef System (GARS), which covers an area of similar to 56,000 km(2) off the Amazon River mouth. Sponges are the major GARS benthic components. However, these sponges' microbiome across the GARS is still unknown. Here, we investigated water quality, isotopic values (delta C-13 and delta N-15), metagenomic and lipidomic profiles of sponges obtained from different sectors throughout the GARS. >180 million shotgun metagenomic reads were annotated, covering 22 sponge species. Isotopic and lipidomic analyses suggested LMA sponges rely on the Amazon River Plume for nutrition. HMA sponges (N = 15) had higher Roseiflexus and Nitrospira abundance, whereas LMA sponges (N= 7) had higher Prochlorococcus and Pelagibacter abundance. 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Proteins involved in methanol oxidation (from the OM43 clade) and carbon monoxide (from a wide variety of bacteria) were identified throughout Bedford Basin. Metabolic niche partitioning between the SUP05 and ARCTIC96BD-19 clades, which together comprise the Gamma-proteobacterial sulfur oxidizers group was apparent. ARCTIC96BD-19 proteins involved in the transport of organic compounds indicated that in productive coastal waters this lineage tends toward a heterotrophic metabolism. In contrast, the identification of sulfur oxidation proteins from SUP05 indicated the use of reduced sulfur as an energy source in hypoxic bottom water. We identified an abundance of Marine Group I Thaumarchaeota proteins in the hypoxic deep layer, including proteins for nitrification and carbon fixation. No transporters for organic compounds were detected among the thaumarchaeal proteins, suggesting a reliance on autotrophic carbon assimilation. 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Kujawinski, Elizabeth B. Longnecker, Krista Lu, YueHan Halewood, Elisa Carlson, Craig A. TI Different carboxyl-rich alicyclic molecules proxy compounds select distinct bacterioplankton for oxidation of dissolved organic matter in the mesopelagic Sargasso Sea SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Marine dissolved organic matter (DOM) varies in its recalcitrance to rapid microbial degradation. DOM of varying recalcitrance can be exported from the ocean surface to depth by subduction or convective mixing and oxidized over months to decades in deeper seawater. Carboxyl-rich alicyclic molecules (CRAM) are characterized as a major component of recalcitrant DOM throughout the oceanic water column. The oxidation of CRAM-like compounds may depend on specific bacterioplankton lineages with oxidative enzymes capable of catabolizing complex molecular structures like long-chain aliphatics, cyclic alkanes, and carboxylic acids. To investigate the interaction between bacteria and CRAM-like compounds, we conducted microbial remineralization experiments using several compounds rich in carboxyl groups and/or alicyclic rings, including deoxycholate, humic acid, lignin, and benzoic acid, as proxies for CRAM. Mesopelagic seawater (200 m) from the northwest Sargasso Sea was used as media and inoculum and incubated over 28 d. All amendments demonstrated significant DOC removal (2-11 mu mol C L-1) compared to controls. Bacterioplankton abundance increased significantly in the deoxycholate and benzoic acid treatments relative to controls, with fast-growing Spongiibacteracea, Euryarcheaota, and slow-growing SAR11 enriched in the deoxycholate treatment and fast-growing Alteromonas, Euryarcheaota, and Thaumarcheaota enriched in the benzoic acid treatment. In contrast, bacterioplankton grew slower in the lignin and humic acid treatments, with oligotrophic SAR202 becoming significantly enriched in the lignin treatment. 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In an effort to culture the uncultured phylotypes from oligotrophic marine ecosystems, we developed high-throughput culturing procedures that utilize the concept of extinction culturing to isolate cultures in small volumes of low-nutrient media. In these experiments, marine bacteria were isolated and cultivated at in situ substrate concentrations-typically 3 orders of magnitude less than common laboratory media. Microtiter plates and a newly developed procedure for making cell arrays were employed to raise the throughput rate and lower detection sensitivity, permitting cell enumeration from 200-mul aliquots of cultures with densities as low as 10(3) cells/ml. Approximately 2,500 extinction cultures from 11 separate samplings of marine bacterio plankton were screened over the course of 3 years. 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Hollins, Ashley Mccarty, Alexandra J. Martinez, Grace Cui, Minming Gomes, Maya L. Fuchsman, Clara A. TI Microbial diversity and abundance vary along salinity, oxygen, and particle size gradients in the Chesapeake Bay SO ENVIRONMENTAL MICROBIOLOGY DT Article AB Marine snow and other particles are abundant in estuaries, where they drive biogeochemical transformations and elemental transport. Particles range in size, thereby providing a corresponding gradient of habitats for marine microorganisms. We used standard normalized amplicon sequencing, verified with microscopy, to characterize taxon-specific microbial abundances, (cells per litre of water and per milligrams of particles), across six particle size classes, ranging from 0.2 to 500 mu m, along the main stem of the Chesapeake Bay estuary. Microbial communities varied in salinity, oxygen concentrations, and particle size. Many taxonomic groups were most densely packed on large particles (in cells/mg particles), yet were primarily associated with the smallest particle size class, because small particles made up a substantially larger portion of total particle mass. However, organisms potentially involved in methanotrophy, nitrite oxidation, and sulphate reduction were found primarily on intermediately sized (5-180 mu m) particles, where species richness was also highest. All abundant ostensibly free-living organisms, including SAR11 and Synecococcus, appeared on particles, albeit at lower abundance than in the free-living fraction, suggesting that aggregation processes may incorporate them into particles. Our approach opens the door to a more quantitative understanding of the microscale and macroscale biogeography of marine microorganisms. Overview of the particle size fractionation approach, and of the microbial community structure of free living bacteria and ones attached to small, intermediate and large particles. Arrows at the bottom indicate that bacterial abundance is highest in the planktonic stages and on smaller particles, and that small particles are more abundance than large ones.image C1 [Cram, Jacob A.; Hollins, Ashley; Mccarty, Alexandra J.; Fuchsman, Clara A.] Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD 21613 USA. [Mccarty, Alexandra J.] Virginia Inst Marine Sci, Marine Advisory Program, Gloucester, VA USA. [Martinez, Grace] Maryland Sea Grant, College Pk, MD USA. [Cui, Minming; Gomes, Maya L.] Johns Hopkins Univ, Earth & Planetary Sci, Baltimore, MD USA. RP Cram, JA (corresponding author), Univ Maryland, Ctr Environm Sci, Horn Point Lab, Cambridge, MD 21613 USA. 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Thingstad, T. Frede Gasol, Josep M. TI Bacterioplankton groups involved in the uptake of phosphate and dissolved organic phosphorus in a mesocosm experiment with P-starved Mediterranean waters SO ENVIRONMENTAL MICROBIOLOGY DT Article AB The use of inorganic phosphate (Pi) and dissolved organic phosphorus (DOP) by different bacterial groups was studied in experimental mesocosms of P-starved eastern Mediterranean waters in the absence (control mesocosms) and presence of additional Pi (P-amended mesocosms). The low Pi turnover times in the control mesocosms and the increase in heterotrophic prokaryotic abundance and production upon Pi addition confirmed that the bacterial community was originally P-limited. The bacterioplankton groups taking up Pi and DOP were identified by means of microautoradiography combined with catalysed reporter deposition fluorescence in situ hybridization. Incubations with leucine were also performed for comparative purposes. All the probe-identified groups showed a high percentage of cells taking up Pi and DOP in the control, P-limited, mesocosms throughout the experiment. However, in response to Pi addition two contrasting scenarios in Pi use were observed: (i) on day 1 of the experiment Pi addition caused a clear reduction in the percentage of SAR11 cells taking up Pi, whereas Gammaproteobacteria, Roseobacter and Bacteroidetes showed similar percentages to the ones in the control mesocosms and (ii) on day 4 of the experiment, probably when the bacterial community had fully responded to the P input, all the probe-identified groups showed low percentages of cells taking up the substrate as compared with the control mesocosms. These differences are likely related to different P requirements among the bacterial groups and point out to the existence of two contrasting strategies in P use. 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Terminal restriction fragment length polymorphisms (T-RFLP) analysis indicated that the HI populations in all four amendments were composed of bacteria from the same major taxa (phylum and subphylum levels), but the relative abundance of each differed. High-resolution clone libraries (each containing similar to 200 clones) showed that the HI populations in the GIyB and VanA amendments consisted of both metabolic generalists and specialists within the alpha-Proteobacteria (mainly members of the Roseobacter clade), beta-Proteobacteria and gamma-Proteobacteria (mainly members of Altermonadaceae, Chromatiaceae, Oceanospirillaceae and Pseudomonadaceae). The presence of members of OM60/241, OM185, SAR11, SAR86 and SAR116 in the HI populations indicated that members of these groups can assimilate the model DOC compounds, providing some of the first glimpses into heterotrophy by members of these poorly understood environmental clusters. C1 Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. 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Rappe, Michael S. TI Non-random assembly of bacterioplankton communities in the subtropical North Pacific Ocean SO FRONTIERS IN MICROBIOLOGY DT Article AB The exploration of bacterial diversity in the global ocean has revealed new taxa and previously unrecognized metabolic potential; however, our understanding of what regulates this diversity is limited. Using terminal restriction fragment length polymorphism (T-RFLP) data from bacterial small-subunit ribosomal RNA genes we show that, independent of depth and time, a large fraction of bacterioplankton co-occurrence patterns are non-random in the oligotrophic North Pacific subtropical gyre (NPSG). Pair-wise correlations of all identified operational taxonomic units (OTUs) revealed a high degree of significance, with 6.6% of the pair-wise co-occurrences being negatively correlated and 20.7% of them being positive. The most abundant OTUs, putatively identified as Prochlorococcus, SAR11, and SAR116 bacteria, were among the most correlated OTUs. As expected, bacterial community composition lacked statistically significant patterns of seasonality in the mostly stratified water column except in a few depth horizons of the sunlit surface waters, with higher frequency variations in community structure apparently related to populations associated with the deep chlorophyll maximum. Communities were structured vertically into epipelagic, mesopelagic, and bathypelagic populations. Permutation-based statistical analyses of T-RFLP data and their corresponding metadata revealed a broad range of putative environmental drivers controlling bacterioplankton community composition in the NPSG, including concentrations of inorganic nutrients and phytoplankton pigments. Together, our results suggest that deterministic forces such as environmental filtering and interactions among taxa determine bacterioplankton community patterns, and consequently affect ecosystem functions in the NPSG. C1 [Eiler, Alexander; Hayakawa, Darin H.; Rappe, Michael S.] Univ Hawaii, Sch Ocean & Earth Sci & Technol, Hawaii Inst Marine Biol, Kaneohe, HI USA. [Eiler, Alexander] Uppsala Univ, Dept Ecol & Genet, Uppsala, Sweden. [Hayakawa, Darin H.] Univ Hawaii, Dept Microbiol, Honolulu, HI 96822 USA. RP Eiler, A (corresponding author), Dept Ecol & Genet, Norbyvagen 18D, SE-75236 Uppsala, Sweden. 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Phillips, Margaret A. Michael, Anthony J. TI Functional polyamine metabolic enzymes and pathways encoded by the virosphere SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Viruses produce more viruses by manipulating the metabolic and replication systems of their host cells. Many have acquired metabolic genes from ancestral hosts and use the encoded enzymes to subvert host metabolism. The polyamine spermidine is required for bacteriophage and eukaryotic virus replication, and herein, we have identified and func-tionally characterized diverse phage-and virus-encoded polyamine metabolic enzymes and pathways. These include pyridoxal 5 '-phosphate (PLP)-dependent ornithine decarboxylase (ODC), pyruvoyl-dependent ODC and arginine decarboxylase (ADC), arginase, S-adenosylmethionine decarboxylase (AdoMetDC/speD), spermidine synthase, homospermidine synthase, spermidine N-acetyltransferase, and N-acetylspermidine amidohydrolase. 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Giske, Jarl Thingstad, T. Frede TI Optimal Defense Strategies in an Idealized Microbial Food Web under Trade-Off between Competition and Defense SO PLOS ONE DT Article AB Trophic mechanisms that can generate biodiversity in food webs include bottom-up (growth rate regulating) and top-down (biomass regulating) factors. The top-down control has traditionally been analyzed using the concepts of "Keystone Predation'' (KP) and "Killing-the-Winner'' (KtW), predominately occuring in discussions of macro- and micro-biological ecology, respectively. Here we combine the classical diamond-shaped food web structure frequently discussed in KP analyses and the KtW concept by introducing a defense strategist capable of partial defense. A formalized description of a trade-off between the defense-strategist's competitive and defensive ability is included. 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Becker, Cynthia Neave, Matthew Hughen, Konrad Dona, Angela Richards Aeby, Greta Work, Thierry Weber, Laura McNally, Sean TI Microbial ecology of coral-dominated reefs in the Federated States of Micronesia SO AQUATIC MICROBIAL ECOLOGY DT Article AB Microorganisms are central to the functioning of coral reef ecosystems, but their dynamics are unstudied on most reefs. We examined the microbial ecology of shallow reefs within the Federated States of Micronesia. We surveyed 20 reefs surrounding 7 islands and atolls (Yap, Woleai, Olimarao, Kosrae, Kapingamarangi, Nukuoro, and Pohnpei), spanning 875 053 km(2). On the reefs, we found consistently higher coral coverage (mean +/- SD = 36.9 +/- 22.2%; max 77 %) compared to macroalgae coverage (15.2 +/- 15.5 %; max 58 %), and low abundances of fish. Reef waters had low inorganic nutrient concentrations and were dominated by Synechococcus, Prochiorococcus, and SAR11 bacteria. The richness of bacterial and archaeal communities was significantly related to interactions between island/atoll and depth. High coral coverage on reefs was linked to higher relative abundances of Flavobacteriaceae, Leisingera, Owenweeksia, Vibrio, and the OM27 clade, as well as other heterotrophic bacterial groups, consistent with communities residing in waters near corals and within coral mucus. Microbial community structure at reef depth was significantly correlated with geographic distance, suggesting that island biogeography influences reef microbial communities. Reefs at Kosrae Island, which hosted the highest coral abundance and diversity, were unique compared to other locations; seawater from Kosrae reefs had the lowest organic carbon (59.8-67.9 mu M), highest organic nitrogen (4.5-5.3 mu M), and harbored consistent microbial communities (>85% similar), which were dominated by heterotrophic cells. This study suggests that the reef-water microbial ecology on Micronesian reefs is influenced by the density and diversity of corals as well as other biogeographical features. C1 [Apprill, Amy; Holm, Henry; Becker, Cynthia; Neave, Matthew; Hughen, Konrad; Weber, Laura; McNally, Sean] Woods Hole Oceanog Inst, Marine Chem & Geochem Dept, Woods Hole, MA 02543 USA. [Holm, Henry; Becker, Cynthia] MIT WHOI Joint Program Oceanog Appl Ocean Sci & E, Cambridge, MA 02139 USA. [Santoro, Alyson E.] Univ Calif Santa Barbara, Dept Ecol Evolut & Marine Biol, Santa Barbara, CA 93106 USA. [Dona, Angela Richards; Aeby, Greta] Univ Hawaii, Honolulu, HI 96822 USA. [Work, Thierry] US Geol Survey, Natl Wildlife Hlth Ctr, Honolulu Field Stn, Honolulu, HI 96850 USA. [Neave, Matthew] CSIRO, Australian Anim Hlth Lab, Geelong, Vic 3219, Australia. [Weber, Laura; McNally, Sean] Univ Massachusetts, Sch Environm, Boston, MA 02125 USA. RP Apprill, A (corresponding author), Woods Hole Oceanog Inst, Marine Chem & Geochem Dept, Woods Hole, MA 02543 USA. 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We tested this hypothesis by conducting 3 light manipulation experiments with bacterial communities obtained off the Southern California coast. In each experiment, duplicate 20 l mesocosms were subjected to either 14:10 h light:dark or continual darkness for 5 to 10 d. Automated Ribosomal Intergenic Spacer Analysis (ARISA), a whole-community fingerprinting technique, was used to determine how light affects bacterial community structure. Light removal resulted in only minor changes in ARISA profiles (mean decrease in similarity between treatments: 6 +/- 1 % via Sorenson's index [only considers presence/absence of taxa], 15 +/- 7 % via Pearson's coefficient [considers relative abundance of taxa]) at the conclusion of the experiments. Oligotrophic communities responded nearly twice as strongly to light removal compared to mesotrophic communities. 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Gregory-Eaves, Irene Walsh, David A. TI Sediment Metagenomes as Time Capsules of Lake Microbiomes SO MSPHERE DT Article AB The reconstruction of ecological time series from lake sediment archives can retrace the environmental impact of human activities. Molecular genetic approaches in paleolimnology have provided unprecedented access to DNA time series, which record evidence of the microbial ecologies that underlaid historical lake ecosystems. Such studies often rely on single-gene surveys, and consequently, the full diversity of preserved microorganisms remains unexplored. In this study, we probed the diversity archived in contemporary and preindustrial sediments by comparative shotgun metagenomic analysis of surface water and sediment samples from three eastern Canadian lakes. In a strategy that was aimed at disentangling historical DNA from the indigenous sediment background, microbial preservation signals were captured by mapping sequence similarities between sediment metagenome reads and reference surface water metagenome assemblies. We detected preserved Cyanobacteria, diverse bacterioplankton, microeukaryotes, and viruses in sediment metagenomes. Among the preserved microorganisms were important groups never before reported in paleolimnological reconstructions, including bacteriophages (Caudoviroles) and ubiquitous freshwater Betaproteobacteria (Polynucleobacter and Limnohabitans). In contrast, ultramicroscopic Actinobacteria ("Candidatus Nanopelagicales") and Alphaproteobacteria (Pelagibacterales) were apparently not well preserved in sediment metagenomes even though they were numerically dominant in surface water metagenomes. Overall, our study explored a novel application of whole-metagenome shotgun sequencing for discovering the DNA remains of a broad diversity of microorganisms preserved in lake sediments. The recovery of diverse microbial time series supports the taxonomic expansion of microbiome reconstructions and the development of novel microbial paleoindicators. IMPORTANCE Lakes are critical freshwater resources under mounting pressure from climate change and other anthropogenic stressors. The reconstruction of ecological time series from sediment archives with paleolimnological techniques has been shown to be an effective means of understanding how humans are modifying lake ecosystems over extended timescales. In this study, we combined shotgun DNA sequencing with a novel comparative analysis of surface water and sediment metagenomes to expose the diversity of microorganisms preserved in lake sediments. The detection of DNA from a broad diversity of preserved microbes serves to more fully reconstruct historical microbiomes and describe preimpact lake conditions. C1 [Garner, Rebecca E.; Walsh, David A.] Concordia Univ, Dept Biol, Montreal, PQ, Canada. [Gregory-Eaves, Irene] McGill Univ, Dept Biol, Montreal, PQ, Canada. [Garner, Rebecca E.; Gregory-Eaves, Irene; Walsh, David A.] Grp Rech Interuniv Limnol, Montreal, PQ, Canada. RP Walsh, DA (corresponding author), Concordia Univ, Dept Biol, Montreal, PQ, Canada.; Walsh, DA (corresponding author), Grp Rech Interuniv Limnol, Montreal, PQ, Canada. 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TI Bacterial Succession on Sinking Particles in the Ocean's Interior SO FRONTIERS IN MICROBIOLOGY DT Article AB Sinking particles formed in the photic zone and moving vertically through the water column are a main mechanism for nutrient transport to the deep ocean, and a key component of the biological carbon pump. The particles appear to be processed by a microbial community substantially different from the surrounding waters. Single cell genomics and metagenomics were employed to describe the succession of dominant bacterial groups during particle processing. Sinking particles were extracted from sediment traps at Station Aloha in the North Pacific Subtropical Gyre (NPSG) during two different trap deployments conducted in July and August 2012. The microbial communities in poisoned vs. live sediment traps differed significantly from one another, consistent with prior observations by Fontanez et al. (2015). Partial genomes from these communities were sequenced from cells belonging to the genus Arcobacter (commensalists potentially associated with protists such as Radiolaria), and Vibrio carnpbellii (a group previously reported to be associated with crustacea). These bacteria were found in the particle-associated communities at specific depths in both trap deployments, presumably due to their specific host-associations. Partial genomes were also sequenced from cells belonging to Idiomarina and Kangiella that were enriched in live traps over a broad depth range, that represented a motile copiotroph and a putatively non-motile algicidal saprophyte, respectively. Planktonic bacterial cells most likely caught in the wake of the particles belonging to Actinomarina and the SAR11 Glade were also sequenced. Our results suggest that similar groups of eukaryote-associated bacteria are consistently found on sinking particles at different times, and that particle remineralization involves specific, reproducible bacterial succession events in oligotrophic ocean waters. C1 [Pelve, Erik A.] Uppsala Univ, Biomed Ctr, Dept Cell & Mol Biol Mol Evolut, Uppsala, Sweden. [Fontanez, Kristina M.] MIT, Dept Civil & Environm Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA. [DeLong, Edward F.] Univ Hawaii Manoa, Dept Oceanog, Daniel K Inoue Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. [Fontanez, Kristina M.] Fluid Screen Inc, Cambridge, MA USA. RP DeLong, EF (corresponding author), Univ Hawaii Manoa, Dept Oceanog, Daniel K Inoue Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. 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Silvano, Eleonora Murphy, Andrew Guillonneau, Richard Quareshy, Mussa Rihtman, Branko Ferretjans, Maria Aguilo He, Ruo Todd, Jonathan D. Chen, Feng Scanlan, David J. Chen, Yin TI Aminolipids elicit functional trade-offs between competitiveness and bacteriophage attachment in Ruegeria pomeroyi SO ISME JOURNAL DT Article AB Lipids play a crucial role in maintaining cell integrity and homeostasis with the surrounding environment. Cosmopolitan marine roseobacter clade (MRC) and SAR11 clade bacteria are unique in that, in addition to glycerophospholipids, they also produce an array of amino acid-containing lipids that are conjugated with beta-hydroxy fatty acids through an amide bond. Two of these aminolipids, the ornithine aminolipid (OL) and the glutamine aminolipid (QL), are synthesized using the O-acetyltransferase OlsA. Here, we demonstrate that OL and QL are present in both the inner and outer membranes of the Gram-negative MRC bacterium Ruegeria pomeroyi DSS-3. In an olsA mutant, loss of these aminolipids is compensated by a concurrent increase in glycerophospholipids. The inability to produce aminolipids caused significant changes in the membrane proteome, with the membrane being less permeable and key nutrient transporters being downregulated while proteins involved in the membrane stress response were upregulated. Indeed, the import of C-14-labelled choline and dimethylsulfoniopropionate, as a proxy for the transport of key marine nutrients across membranes, was significantly impaired in the olsA mutant. Moreover, the olsA mutant was significantly less competitive than the wild type (WT) being unable to compete with the WT strain in co-culture. However, the olsA mutant unable to synthesize these aminolipids is less susceptible to phage attachment. 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However, the number of studies reporting the simultaneous diversity of the phytoplankton and bacterioplankton in Antarctic waters are limited. Here, we report data on the bacterial diversity in relation to phytoplankton community structure in the surface waters of the Ross Sea during the Austral summer 2017. Our results show partially overlapping bacterioplankton communities between the stations located in the Terra Nova Bay (TNB) coastal waters and the Ross Sea Open Waters (RSOWs), with a dominance of members belonging to the bacterial phyla Bacteroidetes and Proteobacteria. In the TNB coastal area, microbial communities were characterized by a higher abundance of sequences related to heterotrophic bacterial genera such as Polaribacter spp., together with higher phytoplankton biomass and higher relative abundance of diatoms. On the contrary, the phytoplankton biomass in the RSOW were lower, with relatively higher contribution of haptophytes and a higher abundance of sequences related to oligotrophic and mixothrophic bacterial groups like the Oligotrophic Marine Gammaproteobacteria (OMG) group and SAR11. We show that the rate of diversity change between the two locations is influenced by both abiotic (salinity and the nitrogen to phosphorus ratio) and biotic (phytoplankton community structure) factors. Our data provide new insight into the coexistence of the bacterioplankton and phytoplankton in Antarctic waters, suggesting that specific rather than random interaction contribute to the organic matter cycling in the Southern Ocean. C1 [Cordone, Angelina; Magliulo, Maria; Bolinesi, Francesco; Selci, Matteo; Giovannelli, Donato; Mangoni, Olga] Univ Naples Federico II, Dept Biol, Naples, Italy. [D'Errico, Giuseppe; Giovannelli, Donato] Polytech Univ Marche, Dept Life Sci, DISVA, Ancona, Italy. [Basili, Marco; de Marco, Rocco; Giovannelli, Donato] Inst Marine Biol Resources & Biotechnol CNR IRBIM, Natl Res Council, Ancona, Italy. [Saggiomo, Maria] Stn Zool Anton Dohrn, Naples, Italy. [Rivaro, Paola] Univ Genoa, Dept Chem & Ind Chem, Genoa, Italy. [Giovannelli, Donato] Rutgers State Univ, Dept Marine & Coastal Sci, New Brunswick, NJ 08901 USA. [Giovannelli, Donato] Woods Hole Oceanog Inst, Dept Marine Chem & Geochem, Woods Hole, MA 02543 USA. [Giovannelli, Donato] Tokyo Inst Technol, Earth Life Sci Inst, Tokyo, Japan. Consorzio Nazl Interuniv Sci Mare CoNISMa, Rome, Italy. [Magliulo, Maria] Univ Essex, Colchester, Essex, England. RP Bolinesi, F; Giovannelli, D (corresponding author), Univ Naples Federico II, Dept Biol, Naples, Italy.; Giovannelli, D (corresponding author), Polytech Univ Marche, Dept Life Sci, DISVA, Ancona, Italy.; Giovannelli, D (corresponding author), Inst Marine Biol Resources & Biotechnol CNR IRBIM, Natl Res Council, Ancona, Italy.; Giovannelli, D (corresponding author), Rutgers State Univ, Dept Marine & Coastal Sci, New Brunswick, NJ 08901 USA.; Giovannelli, D (corresponding author), Woods Hole Oceanog Inst, Dept Marine Chem & Geochem, Woods Hole, MA 02543 USA.; Giovannelli, D (corresponding author), Tokyo Inst Technol, Earth Life Sci Inst, Tokyo, Japan. 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The increasing prevalence of these typically oli-gotrophic taxa may severely impact marine food webs and carbon fluxes by an overall decrease in the efficiency of the biological pump. C1 [Moran, Xose Anxelu G.] King Abdullah Univ Sci & Technol, Div Biol & Environm Sci & Engn, Red Sea Res Ctr, Thuwal 239556900, Saudi Arabia. [Moran, Xose Anxelu G.; Alonso-Saez, Laura; Nogueira, Enrique; Lopez-Urrutia, Angel; Diaz-Perez, Laura; Calvo-Diaz, Alejandra; Arandia-Gorostidi, Nestor; Huete-Stauffer, Tamara M.] Ctr Oceanog Xixon, Inst Espanol Oceanog, Xixon 33212, Asturies, Spain. [Alonso-Saez, Laura] AZTI Tecnalia, Marine Res Div, Sukarrieta 48395, Bizkaia, Spain. [Ducklow, Hugh W.] Columbia Univ, Lamont Doherty Earth Observ, Palisades, NY 10964 USA. [Gonzalez, Natalia] Univ Rey Juan Carlos, Dept Biol & Geol Fis & Quim Inorgan, Madrid 28933, Spain. RP Morán, XAG (corresponding author), King Abdullah Univ Sci & Technol, Div Biol & Environm Sci & Engn, Red Sea Res Ctr, Thuwal 239556900, Saudi Arabia. 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Furthermore, dark DIC-assimilating assemblages were widespread among diverse bacterial classes. Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes dominated the active DIC-assimilating communities across the samples. Actinobacteria, Betaproteobacteria, Deltaproteobacteria, Planctomycetes, and Verrucomicrobia were also implicated in DIC assimilation. Alteromonadales and Oceanospirillales contributed significantly to the DIC-assimilating Gammaproteobacteria within May 2010 clone libraries. 16S rRNA gene sequences related to the sulfur-oxidizing symbionts Arctic96BD-19 were observed in all active DIC assimilating clone libraries. Among the Alphaproteobacteria, clones related to the ubiquitous SAR11 clade were found actively assimilating DIC in all samples. Although not a dominant contributor to our active clone libraries, Betaproteobacteria, when identified, were predominantly comprised of Burkholderia. 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Sun, Shulei Sharma, Shalabh Poretsky, Rachel Moran, Mary Ann TI Transcriptomic analysis of a marine bacterial community enriched with dimethylsulfoniopropionate SO ISME JOURNAL DT Article AB Dimethylsulfoniopropionate (DMSP) is an important source of reduced sulfur and carbon for marine microbial communities, as well as the precursor of the climate-active gas dimethylsulfide (DMS). In this study, we used metatranscriptomic sequencing to analyze gene expression profiles of a bacterial assemblage from surface waters at the Bermuda Atlantic Time-series Study (BATS) station with and without a short-term enrichment of DMSP (25 nM for 30 min). An average of 303 143 reads were obtained per treatment using 454 pyrosequencing technology, of which 51% were potential protein-encoding sequences. 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However, we have little understanding of the response of microbiomes to this pool of anthropogenic dissolved organic carbon (ADOC). In this study, coastal microbial communities were challenged with ADOC at environmentally relevant concentrations. Experiments were performed at two Mediterranean sites with different impact by pollutants and nutrients: off the Barcelona harbor ("BCN"), and at the Blanes Bay ("BL"). ADOC additions stimulated prokaryotic leucine incorporation rates at both sites, indicating the use of ADOC as growth substrate. The percentage of "membranecompromised" cells increased with increasing ADOC, indicating concurrent toxic effects of ADOC. Metagenomic analysis of the BCN community challenged with ADOC showed a significant growth of Methylophaga and other gammaproteobacterial taxa belonging to the rare biosphere. Gene expression profiles showed a taxon-dependent response, with significantly enrichments of transcripts from SAR11 and Glaciecola spp. in BCN and BL, respectively. Further, the relative abundance of transposon-related genes (in BCN) and transcripts (in BL) correlated with the number of differentially abundant genes (in BCN) and transcripts (in BLA), suggesting that microbial responses to pollution may be related to pre-exposure to pollutants, with transposons playing a role in adaptation to ADOC. Our results point to a taxon-specific response to low concentrations of ADOC that impact the functionality, structure and plasticity of the communities in coastal seawaters. This work contributes to address the influence of pollutants on microbiomes and their perturbation to ecosystem services and ocean health. C1 [Cerro-Galvez, Elena; Dachs, Jordi; Fernandez-Pinos, Maria-Carmen; Vila-Costa, Maria] IDAEA CSIC, Dept Environm Chem, Catalunya 08034, Spain. 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De Corte, Daniele Amano, Chie Paliaga, Paolo Ivancic, Ingrid Ortiz, Victor Najdek, Mirjana Herndl, Gerhard J. Sintes, Eva TI Mesozooplankton taurine production and prokaryotic uptake in the northern Adriatic Sea SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Dissolved free taurine, an important osmolyte in phytoplankton and metazoans, has been shown to be a significant carbon and energy source for prokaryotes in the North Atlantic throughout the water column. However, the extent of the coupling between taurine production and consumption over a seasonal cycle has not been examined yet. We determined taurine production by abundant crustacean zooplankton and its role as a carbon and energy source for several prokaryotic taxa in the northern Adriatic Sea over a seasonal cycle. Taurine concentrations were generally in the low nanomolar range, reaching a maximum of 22 nmol L(-1)in fall during aPseudonitzschiabloom and coinciding with the highest zooplankton taurine release rates. Taurine accounted for up to 5% of the carbon, 11% of the nitrogen, and up to 71% of the sulfur requirements of heterotrophic prokaryotes. Members of theRoseobacterclade,Alteromonas, Thaumarchaeota, and Euryarchaeota exhibited higher cell-specific taurine assimilation rates than SAR11 cells. However, cell-specific taurine and leucine assimilation were highly variable in all taxa, suggesting species and/or ecotype specific utilization patterns of taurine and dissolved free amino acids. Copepods were able to cover the bulk taurine requirements of the prokaryotic communities in fall and winter and partly in the spring-summer period. Overall, our study emphasizes the significance of taurine as a carbon and energy source for the prokaryotic community in the northern Adriatic Sea and the importance of crustacean zooplankton as a significant source of taurine and other organic compounds for the heterotrophic prokaryotic community. C1 [Clifford, Elisabeth L.; Amano, Chie; Ortiz, Victor; Herndl, Gerhard J.; Sintes, Eva] Univ Vienna, Dept Funct & Evolutionary Ecol, Vienna, Austria. [De Corte, Daniele] Japan Agcy Marine Earth Sci & Technol JAMSTEC, Res & Dev Ctr Marine Biosci, Yokosuka, Kanagawa, Japan. [Paliaga, Paolo] Juraj Dobrila Univ Pula, Dept Nat & Hlth Sci, Pula, Croatia. [Ivancic, Ingrid; Najdek, Mirjana] Rudjer Boskovic Inst, Ctr Marine Res, Rovinj, Croatia. [Herndl, Gerhard J.] Univ Utrecht, Royal Netherlands Inst Sea Res NIOZ, Dept Marine Microbiol & Biogeochem, Den Burg, Netherlands. [Sintes, Eva] Inst Espanol Oceanog IEO, Ctr Oceanog Baleares, Palma De Mallorca, Spain. RP Sintes, E (corresponding author), Univ Vienna, Dept Funct & Evolutionary Ecol, Vienna, Austria.; Sintes, E (corresponding author), Inst Espanol Oceanog IEO, Ctr Oceanog Baleares, Palma De Mallorca, Spain. 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Maina, Theodosia Orlova, Anna TI GRPR-Antagonists Carrying DOTAGA-Chelator via Positively Charged Linkers: Perspectives for Prostate Cancer Theranostics SO PHARMACEUTICS DT Article AB Gastrin-releasing peptide receptor (GRPR)-antagonists have served as motifs in the development of theranostic radioligands for prostate cancer. Our efforts have been focused on the development of radiolabeled RM26 (H-DPhe6-Gln7-Trp8-Ala9-Val10-Gly11-His12-Sta13-Leu14-NH2) analogs, such as [111In]In-DOTAGA-PEG2-RM26. We recently showed that its Gly11/Sar11-substituted version, [111In]In-AU-RM26-M1, resisted degradation by neprilysin (NEP) while in circulation and achieved higher tumor uptake in mice. We herein introduce the following three new AU-RM26-M1 mimics labeled with In-111, with basic residues in the linker: (i) AU-RM26-M2 (PEG2-Pip), (ii) AU-RM26-M3 (PEG2-Arg), and (iii) AU-RM26-M4 (Arg-Arg-Pip). These analogs were compared in PC-3 cells and animal models vs. AU-RM26-M1 (reference). 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[Kanellopoulos, Panagiotis; Nock, Berthold A.; Maina, Theodosia] NCSR Demokritos, Mol Radiopharm, INRaSTES, Athens 15341, Greece. [Tolmachev, Vladimir] Uppsala Univ, Dept Immunol Genet & Pathol, S-75183 Uppsala, Sweden. [Orlova, Anna] Uppsala Univ, Sci Life Lab, Uppsala, Sweden. RP Orlova, A (corresponding author), Uppsala Univ, Dept Med Chem, S-75183 Uppsala, Sweden.; Orlova, A (corresponding author), Uppsala Univ, Sci Life Lab, Uppsala, Sweden. EM karim.obeid.1838@student.uu.se; panagiotis.kanellopoulos@ilk.uu.se; ayman.abouzayed@ilk.uu.se; adam.mattsson.4918@student.uu.se; vladimir.tolmachev@igp.uu.se; nock_berthold.a@hotmail.com; maina_thea@hotmail.com; anna.orlova@ilk.uu.se CR Aboagye EO, 2023, CA-CANCER J CLIN, V73, P255, DOI 10.3322/caac.21768 Abouzayed A, 2023, BIOMOLECULES, V13, DOI 10.3390/biom13071134 Adessi C, 2002, CURR MED CHEM, V9, P963, DOI 10.2174/0929867024606731 Ananias HJK, 2009, PROSTATE, V69, P1101, DOI 10.1002/pros.20957 Ayalasomayajula S, 2017, CLIN PHARMACOKINET, V56, P1461, DOI 10.1007/s40262-017-0543-3 Azay J, 1998, PEPTIDES, V19, P57, DOI 10.1016/S0196-9781(97)00275-1 Baratto L, 2021, J NUCL MED, V62, P1545, DOI 10.2967/jnumed.120.259630 Baun C, 2024, SEMIN NUCL MED, V54, P256, DOI 10.1053/j.semnuclmed.2024.01.004 Beer M, 2012, PROSTATE, V72, P318, DOI 10.1002/pros.21434 Bodei L, 2022, NAT REV CLIN ONCOL, V19, P534, DOI 10.1038/s41571-022-00652-y Chatalic KLS, 2015, J NUCL MED, V56, P1809, DOI 10.2967/jnumed.115.161158 deCastiglione R, 1996, CRIT REV ONCOL HEMAT, V24, P117, DOI 10.1016/1040-8428(96)00220-X Erak M, 2018, BIOORGAN MED CHEM, V26, P2759, DOI 10.1016/j.bmc.2018.01.012 Fani M, 2011, J NUCL MED, V52, P1110, DOI 10.2967/jnumed.111.087999 Fanti S, 2022, EUR UROL ONCOL, V5, P530, DOI 10.1016/j.euo.2022.05.003 Gandaglia G, 2021, EUR UROL ONCOL, V4, P877, DOI 10.1016/j.euo.2021.09.006 Gotthardt M, 2007, J NUCL MED, V48, P596, DOI 10.2967/jnumed.106.036020 Gu J, 2010, J CLIN PHARMACOL, V50, P401, DOI 10.1177/0091270009343932 Gugger M, 1999, AM J PATHOL, V155, P2067, DOI 10.1016/S0002-9440(10)65525-3 Jensen RT, 2008, PHARMACOL REV, V60, P1, DOI 10.1124/pr.107.07108 Kanellopoulos P, 2020, PHARMACEUTICS, V12, DOI 10.3390/pharmaceutics12121145 Körner M, 2014, PROSTATE, V74, P217, DOI 10.1002/pros.22743 Maina T, 2017, PET CLIN, V12, P297, DOI 10.1016/j.cpet.2017.02.007 Mansi R, 2021, CANCERS, V13, DOI 10.3390/cancers13225766 Mansi R, 2011, EUR J NUCL MED MOL I, V38, P97, DOI 10.1007/s00259-010-1596-9 Markwalder R, 1999, CANCER RES, V59, P1152 Minamimoto R, 2018, J NUCL MED, V59, P803, DOI 10.2967/jnumed.117.197624 Mitran B, 2019, INT J CANCER, V145, P3347, DOI 10.1002/ijc.32401 Morgat C, 2017, J NUCL MED, V58, P1401, DOI 10.2967/jnumed.116.188011 Nock BA, 2023, PHARMACEUTICALS-BASE, V16, DOI 10.3390/ph16050674 Nock BA, 2014, J NUCL MED, V55, P121, DOI 10.2967/jnumed.113.129411 Price EW, 2014, CHEM SOC REV, V43, P260, DOI 10.1039/c3cs60304k REILE H, 1994, PROSTATE, V25, P29, DOI 10.1002/pros.2990250105 Reubi JC, 2004, EUR J NUCL MED MOL I, V31, P803, DOI 10.1007/s00259-004-1476-2 Reubi JC, 2002, CLIN CANCER RES, V8, P1139 ROQUES BP, 1993, BIOCHEM SOC T, V21, P678, DOI 10.1042/bst0210678 ROQUES BP, 1993, PHARMACOL REV, V45, P87 Schreck MV, 2023, MOL PHARMACEUT, V20, P6463, DOI 10.1021/acs.molpharmaceut.3c00852 Sung H, 2021, CA-CANCER J CLIN, V71, P209, DOI 10.3322/caac.21660 Timmis A, 2014, NEW ENGL J MED, V371, P2336, DOI 10.1056/NEJMc1412654 Vlieghe P, 2010, DRUG DISCOV TODAY, V15, P40, DOI 10.1016/j.drudis.2009.10.009 NR 41 TC 4 Z9 4 PD APR PY 2024 VL 16 IS 4 AR 513 DI 10.3390/pharmaceutics16040513 UT WOS:001210289800001 DA 2025-07-30 ER PT J AU Riemann, L Steward, GF Fandino, LB Campbell, L Landry, MR Azam, F AF Riemann, L Steward, GF Fandino, LB Campbell, L Landry, MR Azam, F TI Bacterial community composition during two consecutive NE Monsoon periods in the Arabian Sea studied by denaturing gradient gel electrophoresis (DGGE) of rRNA genes SO DEEP-SEA RESEARCH PART II-TOPICAL STUDIES IN OCEANOGRAPHY DT Article AB Horizontal and vertical variations in bacterial community composition were examined in samples collected during two Joint Global Ocean Flux Study (JGOFS) Arabian Sea cruises in 1995, The cruises, 11 months apart, took place during two consecutive NE Monsoon periods (January and December). Bacteria were harvested by filtration from samples collected in the mixed layer, mid-water, and deep sea at stations across the study area, Total bacterial community genomic DNA was analyzed by PCR amplification of 16S rRNA gene fragments, followed by denaturing gradient gel electrophoresis (DGGE). In total, 20 DGGE bands reflecting unique or varying phylotypes were excised, cloned and sequenced. Amplicons were dominated by bacterial groups commonly found in oceanic waters (e.g,, the SAR11 cluster of alpha-Proteobacteria and cyanobacteria), but surprisingly none of the sequenced amplicons were related to gamma-Proteobacteria or to members of the Cytophaga-Flavobacter-Bacteroides phylum, Amplicons related to magnetotactic bacteria were found for the first time in pelagic oceanic waters. The DGGE banding patterns revealed a dominance of approximate to 15 distinguishable amplicons in all samples. 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(viromes) have been investigated extensively via several large-scale ocean sequencing projects. However, there have not been any systematic viromic studies in estuaries. Here, we investigated the viromes of the Delaware Bay and Chesapeake Bay, two Mid-Atlantic estuaries. Deep sequencing generated a total of 48,190 assembled viral sequences (>5 kb) and 26,487 viral populations (9,204 virus clusters and 17,845 singletons), including 319 circular viral contigs between 7.5 kb and 161.8 kb. Unknown viruses represented the vast majority of the dominant populations, while the composition of known viruses, such as pelagiphage and cyanophage, appeared to be relatively consistent across a wide range of salinity gradients and in different seasons. A difference between estuarine and ocean viromes was reflected by the proportions of Myoviridae, Podoviridae, Siphoviridae, Phycodnaviridae, and a few well-studied virus representatives. The difference in viral community between the Delaware Bay and Chesapeake Bay is significantly more pronounced than the difference caused by temperature or salinity, indicating strong local profiles caused by the unique ecology of each estuary. Interestingly, a viral contig similar to phages infecting Acinetobacter baumannii ("Iraqibacter") was found to be highly abundant in the Delaware Bay but not in the Chesapeake Bay, the source of which is yet to be identified. Highly abundant viruses in both estuaries have close hits to viral sequences derived from the marine single-cell genomes or long-read single-molecule sequencing, suggesting that important viruses are still waiting to be discovered in the estuarine environment. IMPORTANCE This is the first systematic study about spatial and temporal variation of virioplankton communities in estuaries using deep metagenomics sequencing. It is among the highest-quality viromic data sets to date, showing remarkably consistent sequencing depth and quality across samples. Our results indicate that there exists a large pool of abundant and diverse viruses in estuaries that have not yet been cultivated, their genomes only available thanks to single-cell genomics or single -molecule sequencing, demonstrating the importance of these methods for viral discovery. The spatiotemporal pattern of these abundant uncultivated viruses is more variable than that of cultured viruses. Despite strong environmental gradients, season and location had surprisingly little impact on the viral community within an estuary, but we saw a significant distinction between the two estuaries and also between estuarine and open ocean viromes. C1 [Sun, Mengqi; Zhan, Yuanchao; Marsan, David; Chen, Feng] Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA. 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TI Vertical stratification of bacterial communities driven by multiple environmental factors in the waters (0-5000 m) off the Galician coast (NW Iberian margin) SO DEEP-SEA RESEARCH PART I-OCEANOGRAPHIC RESEARCH PAPERS DT Article AB The processes mediated by microbial planktonic communities occur along the entire water column, yet the microbial activity and composition have been studied mainly in surface waters. This research examined the vertical variation in bacterial abundance, activity and community composition and structure from surface down to 5000 m depth following a longitudinal transect off the Galician coast (NW Iberian margin, from 43 degrees N, 9 degrees W to 43 degrees N, 15 degrees W). Community activity and composition changed with depth. The leucine incorporation rates decreased from the euphotic layer to the bathypelagic waters by three orders of magnitude, whereas prokaryotic abundance decreased only by one order of magnitude. 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Fuchs, Bernhard M. TI Phylogenetic characterisation of picoplanktonic populations with high and low nucleic acid content in the North Atlantic Ocean SO SYSTEMATIC AND APPLIED MICROBIOLOGY DT Article AB In flow cytometric analyses of marine prokaryotic picoplankton often two populations with distinct differences in their apparent nucleic acid content are discernable, one with a high and one with a low nucleic acid content (HNA and LNA, respectively). In this study we determined the phylogenetic composition of flow cytometrically sorted HNA and LNA populations, collected at six stations along a transect across three oceanic provinces from Iceland to the Azores. Catalysed reporter deposition fluorescence in situ hybridisation (CARD-FISH) analysis of sorted cells revealed distinct differences in phylogenetic composition between the LNA and HNA populations with only little overlap. At all stations the LNA population was dominated by the alphaproteobacterial clade SAR11 (45-74%). 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Zielinski, Brian L. Doherty, Mary Smith, Christa B. Sharma, Shalabh Paul, John H. Crump, Byron C. Moran, Mary Ann TI The Amazon continuum dataset: quantitative metagenomic and metatranscriptomic inventories of the Amazon River plume, June 2010 SO MICROBIOME DT Article AB Background: The Amazon River is by far the world's largest in terms of volume and area, generating a fluvial export that accounts for about a fifth of riverine input into the world's oceans. Marine microbial communities of the Western Tropical North Atlantic Ocean are strongly affected by the terrestrial materials carried by the Amazon plume, including dissolved (DOC) and particulate organic carbon (POC) and inorganic nutrients, with impacts on primary productivity and carbon sequestration. Results: We inventoried genes and transcripts at six stations in the Amazon River plume during June 2010. At each station, internal standard-spiked metagenomes, non-selective metatranscriptomes, and poly(A)-selective metatranscriptomes were obtained in duplicate for two discrete size fractions (0.2 to 2.0 mu m and 2.0 to 156 mu m) using 150 x 150 paired-end Illumina sequencing. Following quality control, the dataset contained 360 million reads of approximately 200 bp average size from Bacteria, Archaea, Eukarya, and viruses. Bacterial metagenomes and metatranscriptomes were dominated by Synechococcus, Prochlorococcus, SAR11, SAR116, and SAR86, with high contributions from SAR324 and Verrucomicrobia at some stations. Diatoms, green picophytoplankton, dinoflagellates, haptophytes, and copepods dominated the eukaryotic genes and transcripts. Gene expression ratios differed by station, size fraction, and microbial group, with transcription levels varying over three orders of magnitude across taxa and environments. 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Cluster 1 taxa were characterized by gene expression for motility and complex carbohydrate degradation (dominated by Gammaproteobacteria and Bacteroidetes), and Cluster 2 taxa by transcription of genes for amino acid and aromatic compound metabolism and aerobic anoxygenic phototrophy (Roseobacter). Other activity clusters were enriched in transcripts for proteorhodopsin and methylotrophy (Cluster 4; SAR11 and methylotrophs), photosynthesis and attachment (Clusters 5 and 7; Synechococcus, picoeukaryotes, Verucomicrobia, and Planctomycetes), and sulfur oxidation (Cluster 7; Gammaproteobacteria). The seasonal patterns in activity were overlain, and sometimes obscured, by large differences in %RP over shorter day-night timescales. Seventy-eight taxa, many of them heterotrophs, had a higher %RP activity index during the day than night, indicating a strong diel activity rhythm at this coastal site. 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We hypothesized a direct effect of vents on prokaryotic community structure and functioning in the Panarea Island's hydrothermal system. Sampling was conducted along a 9-station transect characterized by three active emission points. The water column was stratified with a thermocline at 25 m depth and a deep chlorophyll maximum between 50 and 100 m. Prokaryotic abundance ranged from 0.2 to 1.5 x 10(9) cells L-1, prokaryotic carbon production from 2.4 to 75.4 ng C L-1 h(-1), and exoenzymatic activities degrading proteins, phosphorylated compounds, and polysaccharides were on the order of 4-28, 2-31 and 0.2-4.16 nM h(-1), respectively. While microbial abundance and production were shaped by the water column's physical structure, alkaline phosphatase and beta-glucosidase activities seemed to be enhanced by hydrothermal fluids. The 16S rRNA gene amplicon sequencing analysis identified a surface, a deep, and a vent-influenced microbial community. 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Memery, Laurent Paillard, Christine Maignien, Lois TI Linking Spatial and Temporal Dynamic of Bacterioplankton Communities With Ecological Strategies Across a Coastal Frontal Area SO FRONTIERS IN MARINE SCIENCE DT Article AB Ocean frontal systems are widespread hydrological features defining the transition zone between distinct water masses. They are generally of high biological importance as they are often associated with locally enhanced primary production by phytoplankton. However, the composition of bacterial communities in the frontal zone remains poorly understood. In this study, we investigate how a coastal tidal front in Brittany (France) structures the free-living bacterioplankton communities in a spatio-temporal survey across four cruises, five stations and three depths. 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Picoplankton populations collected from the pycnocline (10 m) over the eastern continental shelf of the United States near Cape Hatteras, North Carolina, served as the source of bulk nucleic acids used in this study. A large proportion of the 169 rDNA clones recovered (33%) were related to plastid 16S rRNA genes, including plastids from both chromophyte and chlorophyte algae. Most bacterial gene clones (75% of bacterial clones, 50% of the total) were closely related to rRNA gene lineages that had been discovered previously in clone libraries from open-ocean marine habitats, including the SAR86 cluster (gamma-Proteobacteria), SAR83, SAR11, and SAR116 clusters (all or-Proteobacteria), as well as the marine Gram-positive cluster (high G+C Gram-positive). Most of the remaining bacterial clones recovered were phylogenetically related to the gamma and beta subclasses of the Proteobacteria, including an rDNA lineage within the type 1 methylotroph clade of the beta subclass. 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In productive coastal areas, the biomass of bacterioplankton can be in the same range as that of phytoplankton. In these coastal areas the initiation and intensity of the spring bloom is highly variable between years. This variability is reflected in higher trophic levels and is therefore of major importance for ecosystems such as that of the Faroe Islands. However, one of the major unknown components is the bacterioplankton. We report a study on seasonal dynamics from March-September of nutrients, phytoplankton composition and their co-fluctuation with bacterial succession. For this purpose SAR11, Bacteroidetes, Roseobacter and cyanobacteria were relatively quantified using real-time PCR based on 16S DNA and total bacteria was assessed by epifluorescence microscopy. The phytoplankton species were identified using the inverted microscope technique. These data showed a pronounced diatom spring bloom and autumn bloom reflected by a corresponding decrease in nitrate and silicate (R-2 = 0.72 and 0.77, respectively). The cessation of the phytoplankton bloom did not, however, seem to be explained by nutrient limitation. Roseobacter bloomed during the phytoplankton spring bloom, while the other bacterial groups increased during low phytoplankton biomass. This suggests that algal substrate availability and environmental conditions provide the opportunity for bacterial communities to develop a post-spring bloom. This study reveals how planktonic bacteria adapt with their surroundings, enhancing the microbial loop post-spring bloom and providing a potentially important food resource for higher trophic levels. C1 [Djurhuus, Anni; Jorgensen, Johanna; Hatun, Hjalmar; Debes, Hogni Hammershaimb] Faroe Marine Res Inst, FO-100 Torshavn, Faroe Islands, Denmark. 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Smith, David C. Sogin, Mitchell L. D'Hondt, Steven TI Bacterial and archaeal biogeography of the deep chlorophyll maximum in the South Pacific Gyre SO AQUATIC MICROBIAL ECOLOGY DT Article AB We used 16S rRNA gene tag pyro sequencing to examine the biogeography of bacterial and archaeal community composition in the deep chlorophyll maximum (DCM) of the South Pacific Gyre (SPG), the largest and most oligotrophic region of the world ocean. Dominant DCM bacterial taxa, including Prochlorococcus, SAR11, SAR406, and SAR86, were present at each sampled site in similar proportions, although the sites are separated by thousands of kilometers and up to 100 m in water depth. Marine Group II (MGII) and MGIII Euryarcheota dominated the archaeal assemblages of the DCM at these sites. Bray-Curtis indices show that assemblage composition of these sites is > 70% similar for Bacteria and > 80% similar for Archaea. Despite these similarities, communities of the central SPG, the western SPG margin, and the southern SPG margin are distinguishable from each other. Comparison of our bacterial results to samples from the DCM of the North Pacific Gyre (NPG) and the relatively nutrient-and chlorophyll-rich Equatorial Pacific (EQP) shows that DCM bacterial assemblage composition is > 50% similar throughout all 3 regions. Nonetheless, the SPG, NPG, and EQP assemblages are statistically distinct from each other (ANOSIM, p = 0.001), with the communities of the 2 gyres resembling each other more closely than either resembles the EQP community (which lives geographically between them). Variation in assemblage composition correlates with sea-surface chlorophyll concentration (r(2) = 0.71, p < 0.003). This study demonstrates that the DCM horizons of different oceanic regions harbor statistically distinct communities that are consistent within regions for thousands of kilometers. 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Abraham, Paul E. Shah, Manesh Verberkmoes, Nathan C. Smith, Daniel P. Barofsky, Douglas F. Giovannoni, Stephen J. TI Environmental proteomics of microbial plankton in a highly productive coastal upwelling system SO ISME JOURNAL DT Article AB Metaproteomics is one of a suite of new approaches providing insights into the activities of microorganisms in natural environments. Proteins, the final products of gene expression, indicate cellular priorities, taking into account both transcriptional and posttranscriptional control mechanisms that control adaptive responses. Here, we report the proteomic composition of the < 1.2 mu m fraction of a microbial community from Oregon coast summer surface waters, detected with two-dimensional liquid chromatography coupled with electrospray tandem mass spectrometry. Spectra corresponding to proteins involved in protein folding and biosynthesis, transport, and viral capsid structure were the most frequently detected. A total of 36% of all the detected proteins were best matches to the SAR11 clade, and other abundant coastal microbial clades were also well represented, including the Roseobacter clade (17%), oligotrophic marine gammaproteobacteria group (6%), OM43 clade (1%). Viral origins were attributed to 2.5% of proteins. In contrast to oligotrophic waters, phosphate transporters were not highly detected in this nutrient-rich system. However, transporters for amino acids, taurine, polyamines and glutamine synthetase were among the most highly detected proteins, supporting predictions that carbon and nitrogen are more limiting than phosphate in this environment. Intriguingly, one of the highly detected proteins was methanol dehydrogenase originating from the OM43 clade, providing further support for recent reports that the metabolism of one-carbon compounds by these streamlined methylotrophs might be an important feature of coastal ocean biogeochemistry. The ISME Journal (2011) 5, 856-865; doi:10.1038/ismej.2010.168; published online 11 November 2010 C1 [Giovannoni, Stephen J.] Oregon State Univ, Dept Microbiol, Corvallis, OR 97331 USA. [Sowell, Sarah M.; Smith, Daniel P.] Oregon State Univ, Mol & Cellular Biol Program, Corvallis, OR 97331 USA. [Abraham, Paul E.; Shah, Manesh; Verberkmoes, Nathan C.] Oak Ridge Natl Lab, Div Chem, Oak Ridge, TN 37831 USA. [Abraham, Paul E.; Shah, Manesh; Verberkmoes, Nathan C.] Oak Ridge Natl Lab, Biosci Div, Oak Ridge, TN 37831 USA. [Abraham, Paul E.] Univ Tennessee, Grad Sch Genome Sci & Technol, Knoxville, TN USA. [Barofsky, Douglas F.] Oregon State Univ, Dept Chem, Corvallis, OR 97331 USA. RP Giovannoni, SJ (corresponding author), Oregon State Univ, Dept Microbiol, 220 Nash Hall, Corvallis, OR 97331 USA. 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Gonzalez-Serrano, Rafael Parras-Molto, Marcos Rodriguez-Valera, Francisco TI Genome diversity of marine phages recovered from Mediterranean metagenomes: Size matters SO PLOS GENETICS DT Article AB Marine viruses play a critical role not only in the global geochemical cycles but also in the biology and evolution of their hosts. Despite their importance, viral diversity remains under-explored mostly due to sampling and cultivation challenges. Direct sequencing approaches such as viromics has provided new insights into the marine viral world. As a complementary approach, we analysed 24 microbial metagenomes (> 0.2 mu m size range) obtained from six sites in the Mediterranean Sea that vary by depth, season and filter used to retrieve the fraction. Filter-size comparison showed a significant number of viral sequences that were retained on the larger-pore filters and were different from those found in the viral fraction from the same sample, indicating that some important viral information is missing using only assembly from viromes. Besides, we were able to describe 1,323 viral genomic fragments that were more than 10Kb in length, of which 36 represented complete viral genomes including some of them retrieved from a cross-assembly from different metagenomes. Host prediction based on sequence methods revealed new phage groups belonging to marine prokaryotes like SAR11, Cyanobacteria or SAR116. We also identified the first complete vir-ophage from deep seawater and a new endemic clade of the recently discovered Marine group II Euryarchaeota virus. Furthermore, analysis of viral distribution using metagenomes and viromes indicated that most of the new phages were found exclusively in the Mediterranean Sea and some of them, mostly the ones recovered from deep metagenomes, do not recruit in any database probably indicating higher variability and endemicity in Mediterranean bathypelagic waters. Together these data provide the first detailed picture of genomic diversity, spatial and depth variations of viral communities within the Mediterranean Sea using metagenome assembly. C1 [Lopez-Perez, Mario; Haro-Moreno, Jose M.; Gonzalez-Serrano, Rafael; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Div Microbiol, Evolutionary Genom Grp, Campus San Juan, Alacant, Spain. [Parras-Molto, Marcos] CSIC, Ctr Biol Mol Severo Ochoa, Madrid, Spain. [Parras-Molto, Marcos] Univ Autonoma Madrid, Madrid, Spain. 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AU Zhang, XL Wang, L Peng, SJ Wuzhong, QY Zhang, L Dong, ZJ Sun, TT Liu, P Zhang, QQ Zhao, JN AF Zhang Xiaoli Wang Lei Peng Saijun Wuzhong Qiyue Zhang Lei Dong Zhijun Sun Tingting Liu Ping Zhang Qianqian Zhao Jiamnin TI Abundance, Diversity and Functional Potentials of Planktonic Bacteria and Microeukaryotes in the Coral-Reef System of Xisha Islands, China SO JOURNAL OF OCEAN UNIVERSITY OF CHINA DT Article AB Corals influence microorganisms within the surrounding seawater, yet the diversities and functions of seawater bacteria and microeukaryotes in coral-reef systems have not been well addressed. We collected 40 seawater samples in outer coral reef flats and semi-closed inner lagoons from the surface, middle and bottom layers in the pristine coral-reef system of Xisha Islands, South China Sea. We detected the abundance, composition and distribution of bacteria and microeukaryotes using flow cytometry, qPCR and high throughput sequencing techniques, and profiled the potential ecological roles based on the information of 16S and 18S rDNA sequencing. In terms of flow cytometry, Prochlorococcus dominated the autotrophs with cell abundance ranging from 5.8 x 10(2) to 5.44 x 10(3) cells mL(-1) seawater. Based on qPCR, the 16S rDNA copies were much higher in coral reef flats than in lagoons (P=0.003). The bacterial communities held significantly lower diversity in bottom waters compared with surface and middle waters (P < 0.05), which were dominated by SAR11, Flavobacteriales, and Synechococcus. 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Gray, Patrick C. Niebergall, Alexandria K. Johnson, Zackary I. Hunt, Dana E. TI A Gulf Stream frontal eddy harbors a distinct microbiome compared to adjacent waters SO PLOS ONE DT Article AB Mesoscale oceanographic features, including eddies, have the potential to alter productivity and other biogeochemical rates in the ocean. Here, we examine the microbiome of a cyclonic, Gulf Stream frontal eddy, with a distinct origin and environmental parameters compared to surrounding waters, in order to better understand the processes dominating microbial community assembly in the dynamic coastal ocean. Our microbiome-based approach identified the eddy as distinct from the surround Gulf Stream waters. The eddy-associated microbial community occupied a larger area than identified by temperature and salinity alone, increasing the predicted extent of eddy-associated biogeochemical processes. While the eddy formed on the continental shelf, after two weeks both environmental parameters and microbiome composition of the eddy were most similar to the Gulf Stream, suggesting the effect of environmental filtering on community assembly or physical mixing with adjacent Gulf Stream waters. In spite of the potential for eddy-driven upwelling to introduce nutrients and stimulate primary production, eddy surface waters exhibit lower chlorophyll a along with a distinct and less even microbial community, compared to the Gulf Stream. At the population level, the eddy microbiome exhibited differences among the cyanobacteria (e.g. lower Trichodesmium and higher Prochlorococcus) and in the heterotrophic alpha Proteobacteria (e.g. lower relative abundances of specific SAR11 phylotypes) versus the Gulf Stream. However, better delineation of the relative roles of processes driving eddy community assembly will likely require following the eddy and surrounding waters since inception. 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This may have severe repercussions on phytoplankton bloom dynamics and the associated cycling of carbon in surface waters. We currently lack baseline knowledge of the seasonal dynamics of Arctic microbial communities, which is needed in order to better estimate the effects of such changes on ecosystem functioning. Here we present a comparative study of polar summer microbial communities in the ice-free (eastern) and ice-covered (western) hydrographic regimes at the LTER HAUSGARTEN in Fram Strait, the main gateway between the Arctic and North Atlantic Oceans. Based on measured and modeled biogeochemical parameters, we tentatively identified two different ecosystem states (i.e., different phytoplankton bloom stages) in the distinct regions. Using lllumina tag-sequencing, we determined the community composition of both free-living and particle-associated bacteria as well as microbial eukaryotes in the photic layer. Despite substantial horizontal mixing by eddies in Fram Strait, pelagic microbial communities showed distinct differences between the two regimes, with a proposed early spring (pre-bloom) community in the ice-covered western regime (with higher representation of SAR11, SAR202, SAR406 and eukaryotic MALVs) and a community indicative of late summer conditions (post-bloom) in the icefree eastern regime (with higher representation of Flavobacteria, Gammaproteobacteria and eukaryotic heterotrophs). Co-occurrence networks revealed specific taxon-taxon associations between bacterial and eukaryotic taxa in the two regions. Our results suggest that the predicted changes in sea ice cover and phytoplankton bloom dynamics will have a strong impact on bacterial community dynamics and potentially on biogeochemical cycles in this region. 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Bioinformatic analysis rendered a total of 143600 high quality sequences with an average 11967 sequences per sample and mean read length of 449 bp. Phylogenetic analysis showed that Proteobacteria dominated in all shallow and deep waters, with Alphaproteobacteria and Gammaproteobacteria the two most abundant components, and SAR11 the most abundant group at family level in all regions. Cyanobacteria occurred mainly in the surface euphotic layer, and the majority of them in the tropical waters belonged to the GpIIa family including Prochlorococcus and Synechococcus, whilst those associated with Cryptophytes and diatoms were common in the subarctic waters. In general, species richness (Chao1) and diversity (Shannon index H') were higher for the bacterial communities in the intermediate water layers than for those in surface and deep waters. 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To gain deeper understanding of variations in marine microbial biodiversity and function with inorganic nutrient loading, a large-scale mesocosm (1,000 L) experiment was conducted in southern KCW for 10 days in summer (July 27 through August 6, 2015) and the bacterioplankton community composition (BCC) was investigated using 16S rRNA amplicon pyrosequencing method. The bioassays established in the mesocosm were designed with the following each nutrient dosing treatments: nitrate (+N), phosphate (+P), and nitrate plus phosphate (+NP). Among the three treatments, the +NP and +N treatments exhibited the largest and second-largest increases in phytoplankton abundance, respectively, whereas there was relatively less variation under the +P treatment. Enhanced growth of phytoplankton not only induced increases in pH and bacterial abundance (p < .05) but also led to changes in the BCC: the orders Flavobacteria, Micrococcales, Oceanospirillales, and Rhodobacterales exhibited increases in relative abundance, whereas Methylophilales, Puniceicoccales, SAR11, SAR116, and SAR86 showed a decreasing trend. Inorganic nutrients may directly contribute to variation in the BCC, as the relative abundance of Sphingobacteriales, containing members that undergo chemotaxis toward inorganic nutrients, was increased by the +N treatment. Given that these findings, changes in phytoplankton abundance due to nutrient addition may be the most crucial component resulting in variation in the BCC. C1 [Park, Bum Soo] Korea Inst Ocean Sci & Technol, Marine Ecosyst Res Ctr, Busan, South Korea. [Lee, Minji; Baek, Seung Ho] Korea Inst Ocean Sci & Technol, Risk Assessment Res Ctr, Geoje 53201, South Korea. 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In early spring and summer, these diatomdominated blooms are associated with distinct heterotrophic prokaryotic communities, but whether a structuring effect extends to the post-bloom period has thus far not been investigated. To address this question, we carried out a detailed study of the prokaryotic community composition in the region of Kerguelen Island during late Austral summer (18 February to 27 March 2018; MOBYDICK project). Concentrations of chlorophyll a were seasonally low above the plateau (0.27-0.58 mu g Chl a L-1) and in a similar range to those at the 3 off-plateau sites investigated (0.14-0.34 mu g Chl a L-1), but we observed an accumulation of dissolved organic carbon and the build-up of heterotrophic prokaryotic biomass in Kerguelen plateau waters. Illumina sequencing of the 16S rRNA gene revealed that the total (DNA-based) and potentially active (RNA-based) prokaryotic communities were structured according to on- and off-plateau sites in the wind-mixed surface layer, in both the free-living ( 0.8 mu m size fraction) and particle-attached ( 0.8 mu m size fraction) fractions. The Amplicon Sequence Variants (ASV) with significantly higher relative abundances in on-plateau surface waters as compared to off-plateau waters belonged to Halieaceae OM60 group, several Flavobacteriaceae, such as the NS5 marine group, Aurantivirga and Ulvibacter, Rhodobacteraceae Loktanella, Saprospiraceae, and the Cryomorphaceae NS10 marine group. ASVs with higher relative abundances in off-plateau waters belonged to the Flavobacteriaceae Formosa, the Rhodobacteraceae Planktomarina and the SAR11 clade. We discuss the potential abiotic and biotic drivers of community composition in late Austral summer and the ecological roles of abundant prokaryotic taxa in Kerguelen plateau waters. C1 [Hernandez-Magana, Alejandra Elisa; Liu, Yan; Debeljak, Pavla; Crispi, Olivier; Marie, Barbara; Koedooder, Coco; Obernosterer, Ingrid] Sorbonne Univ, CNRS, Lab Oceanog Microbienne, LOMIC, F-66650 Banyuls Sur Mer, France. [Hernandez-Magana, Alejandra Elisa] Univ Southern Denmark, Dept Biol, Nordcee, Odense M, Denmark. [Liu, Yan] Ludong Univ, Sch Life Sci, Yantai, Peoples R China. [Debeljak, Pavla] Univ Vienna, Dept Funct & Evolutionary Ecol, A-1090 Vienna, Austria. [Debeljak, Pavla] Sorbonne Univ, CNRS, Museum Natl Hist Nat, Inst Syst,Evolut,Biodiversite ISYEB, Banyuls Sur Mer, France. [Debeljak, Pavla; Koedooder, Coco] Hebrew Univ Jerusalem, Fredy & Nadine Herrmann Inst Earth Sci, Jerusalem, Israel. RP Obernosterer, I (corresponding author), Sorbonne Univ, CNRS, Microbial Oceanog Lab LOMIC, 1 Ave Pierre Fabre, F-66650 Banyuls Sur Mer, France. 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Methane was detected throughout the water column; its concentration in the surface water was an order of magnitude lower than in the near-bottom water and 4 orders of magnitude lower than in the sediments. Cyanobacteria of the species Microcystisaeruginosa predominated in the photic zone (up to 30% of the total number of the 16S rRNA gene fragments). The sequences of cyanobacteria and freshwater members of the SAR11 clade, which can potentially be involved in aerobic methanogenesis via decomposition of methylphosphonates (MPn), were also detected. The sequences of hydrogenotrophic methanogens of the genus Methanoregula, which are potentially capable of methanogenesis in cooperation with cyanobacteria, were revealed in oxygen-supersaturated water. Hydrogenotrophic and acetoclastic pathways of methanogenesis predominated in reduced sediments. Sequences of methanogens of the orders Methanomicrobiales, Methanobacteriales, Methanosarciniales, and Methanomassiliicoccales were detected there. Cyanobacterial bloom promoted methanogenesis both in the photic zone of Lake Senezh (due to MPn decomposition and anaerobic methanogenesis in association with cyanobacterial aggregates) and in the near-bottom water and sediments (due to oxygen depletion and excessive release of substrates caused by sedimentation and degradation of cyanobacterial mortmass). C1 [Kallistova, A. Yu.; Kosyakova, A. I.; Rusanov, I. I.; Kadnikov, V. V.; Beletsky, A. V.; Koval, D. D.; Yusupov, S. K.; Pimenov, N. V.] Russian Acad Sci, Fed Res Ctr Biotechnol, Moscow 119071, Russia. [Zekker, I.] Univ Tartu, EE-50411 Tartu, Estonia. RP Kallistova, AY (corresponding author), Russian Acad Sci, Fed Res Ctr Biotechnol, Moscow 119071, Russia. 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On the northern slope of the South China Sea (SCS), a considerable reservoir of mesopelagic fishes that present diurnal vertical movement (DVM) as a diel habit has been discovered. To investigate the microbial community and its potential effects on nutrient conversion in this mesopelagic fish habitat, samples were collected throughout the water column every 6 h to monitor changes over the diel cycle and subjected to high-throughput sequencing of the bacterial and archaeal 16S rRNA genes. The bacterial diversity was high and stratified. Most of the bacteria were R-strategists, which indicates strong dynamics. The dominant bacteria were aerobic or facultatively anaerobic chemo-organotrophs, e.g. Flavobacteriaceae, SAR11, Alteromonadales, and Pseudomonadales, whereas Cyanobacteria was detected throughout the water column in rather low abundances. In deeper layers ( >= 200 m), Deferribacterales, which are typical nitrate reducers, increased in abundance after fish digestion and defecation, were correlated with the DVM. The archaeal diversity was extremely low in areas where Marine Group I appeared as the absolute dominant group throughout the entire water column, and this group presents a high ammonium oxidizing potential. The microbial community structural profile suggested that bacteria and archaea played separate roles in the nitrogen cycling process, which was correlated with the fish activity. Moreover, the protein digesters became abnormally more abundant in the deep water at night, which was likely stimulated by the fecal matter and detritus from fish. Our results provide insights into the spatial and temporal distribution of the microbial communities in the SCS, particularly in relation to mesopelagic fish, and thus will be of assistance for estimating the ecological impact of these communities on SCS fish and fisheries based on the enhanced carbon export that occurs via the diel vertical migration of mesopelagic fish. C1 [Zhang, Yu; Lu, Ye; Wang, Jiahua; Xu, Lei; Xiao, Xiang; Xu, Jun] Shanghai Jiao Tong Univ, Sch Oceanog, State Key Lab Ocean Engn, Shanghai 200240, Peoples R China. [Zhang, Yu; Lu, Ye; Wang, Jiahua; Xie, Lisa; Xu, Lei; He, Ying; Xiao, Xiang; Xu, Jun] Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China. RP Xu, J (corresponding author), Shanghai Jiao Tong Univ, Sch Oceanog, State Key Lab Ocean Engn, Shanghai 200240, Peoples R China. 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TI Intraclade Heterogeneity in Nitrogen Utilization by Marine Prokaryotes Revealed Using Stable Isotope Probing Coupled with Tag Sequencing (Tag- SIP) SO FRONTIERS IN MICROBIOLOGY DT Article AB Nitrogen can greatly influence the structure and productivity of microbial communities through its relative availability and form. However, the roles of specific organisms in the uptake of different nitrogen species remain poorly characterized. Most studies seeking to identify agents of assimilation have been correlative, indirectly linking activity measurements (e.g., nitrate uptake) with the presence or absence of biological markers, particularly functional genes and their transcripts. Evidence is accumulating of previously underappreciated functional diversity in major microbial subpopulations, which may confer physiological advantages under certain environmental conditions leading to ecotype divergence. This microdiversity further complicates our view of genetic variation in environmental samples requiring the development of more targeted approaches. Here, next-generation tag sequencing was successfully coupled with stable isotope probing (Tag-SIP) to assess the ability of individual phylotypes to assimilate a specific N source. Our results provide the first direct evidence of nitrate utilization by organisms thought to lack the genes required for this process including the heterotrophic clades SAR11 and the Archaeal Marine Group II. Alternatively, this may suggest the existence of tightly coupled metabolisms with primary assimilators, e.g., symbiosis, or the rapid and efficient scavenging of recently released products by highly active individuals. These results may be connected with global dominance often seen with these clades, likely conferring an advantage over other clades unable to access these resources. 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To explore how the macroenvironment (environmental filtering) and species-engineered microenvironment (niche construction) affect the ecological network of the marine microbiome in the highly dynamic coastal waters of Korea, we analyzed the modular structures of the microbial community and identified microbial interconnections in different size fractions for a year. Fluctuations in the macroenvironment, such as temperature and nutrient concentrations driven by seasonal changes, are the major factors in determining successive microbial modules. Compared to particle-associated (PA) microbes, free-living (FL) microbes seemed to be more affected by macroenvironmental filtering. Modules related to nutrients were further divided into various modules according to different lifestyles. 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In conclusion, both macro-and micro-environmental filtering were important for determining the seasonal succession of microbial community structures. (c) 2021 Elsevier B.V. All rights reserved. C1 [Chun, Seong-Jun] Natl Inst Ecol, LMO Res Team, 1210 Geumgang Ro, Maseo Myeon 33657, Seocheon, South Korea. [Chun, Seong-Jun; Cui, Yingshun; Ahn, Chi-Yong; Oh, Hee-Mock] Korea Res Inst Biosci & Biotechnol KRIBB, Cell Factory Res Ctr, 125 Gwahak Ro, Daejeon 34141, South Korea. [Baek, Seung Ho] Korea Inst Ocean Sci & Technol, South Sea Inst, Geoje 53201, South Korea. [Ahn, Chi-Yong; Oh, Hee-Mock] Korea Univ Sci & Technol UST, KRIBB Sch Biotechnol, Dept Environm Biotechnol, 217 Gajeong Ro, Daejeon 34113, South Korea. RP Ahn, CY; Oh, HM (corresponding author), Korea Res Inst Biosci & Biotechnol KRIBB, Cell Factory Res Ctr, 125 Gwahak Ro, Daejeon 34141, South Korea. 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J. Treusch, Alexander H. TI Effects of ecological engineered oxygenation on the bacterial community structure in an anoxic fjord in western Sweden SO ISME JOURNAL DT Article AB Oxygen-depleted bodies of water are becoming increasingly common in marine ecosystems. Solutions to reverse this trend are needed and under development, for example, by the Baltic deep-water OXygenation (BOX) project. In the framework of this project, the Swedish Byfjord was chosen for a pilot study, investigating the effects of an engineered oxygenation on long-term anoxic bottom waters. The strong stratification of the water column of the Byfjord was broken up by pumping surface water into the deeper layers, triggering several inflows of oxygen-rich water and increasing oxygen levels in the lower water column and the benthic zone up to 110 mu mol l(-1). We used molecular ecologic methods to study changes in bacterial community structure in response to the oxygenation in the Byfjord. 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C1 [Forth, Michael; Treusch, Alexander H.] Univ Southern Denmark, Nord Ctr Earth Evolut, Dept Biol, DK-5230 Odense M, Denmark. [Liljebladh, Bengt; Stigebrandt, Anders] Univ Gothenburg, Dept Earth Sci, Gothenburg, Sweden. [Hall, Per O. J.] Univ Gothenburg, Dept Chem & Mol Biol, Gothenburg, Sweden. RP Treusch, AH (corresponding author), Univ Southern Denmark, Dept Biol, Campusvej 55, DK-5230 Odense M, Denmark. 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Bosch, Rafael Nogales, Balbina TI Short-term changes in the composition of active marine bacterial assemblages in response to diesel oil pollution SO MICROBIAL BIOTECHNOLOGY DT Article AB The changes caused by diesel oil pollution in the metabolically active bacterioplankton from an oligotrophic coastal location were analysed in laboratory microcosms (44 l) using 16S ribosomal RNA (16S rRNA) as molecular marker. The aim was to simulate typical hydrocarbon pollution events in a coastal area exploited for seasonal touristic activities. The experiment consisted in addition of low amounts of diesel oil without nutrients to seawater collected at different times (winter and summer). Bacterial diversity was analysed by terminal-restriction fragment length polymorphism (T-RFLP) profiling of 16S rRNAs after reverse transcription polymerase chain reaction (RTPCR), and by generation of 16S rRNA clone libraries in control and diesel-polluted microcosms. Diesel addition caused a twofold increase in prokaryotic numbers in comparison with controls at the end of the experiment, both in winter and summer microcosms. Bacterioplankton composition, determined by 16S rRNA T-RFLP data, changed rapidly (within 17 h) in response to treatment. The resulting communities were different in microcosms with water collected in summer and winter. A reduction in diversity (Shannon index, calculated on the basis of T-RFLP data) was observed only in summer microcosms. This was due to the rapid increase of phylotypes affiliated to the Oceanospirillaceae, not observed in winter microcosms. After diesel treatment there was a reduction in the number of phylotypes related to SAR11, SAR86 and picocyanobacteria, while phylotypes of the Roseobacter clade, and the OMG group seemed to be favoured. 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Furthermore, in Flavobacteriales, Alteromonadales, Pelagibacterales, and Rhodobacterales, NCF potential was positively correlated with proton-pump rhodopsin (PPR) expression, suggesting that NCF might be energetically supported by PPR. The novel insights into the lineage-differential potential of carbon fixation, widespread mixotrophy, and PPR as an energy source for NCF lay a methodological and informational foundation for further research to understand carbon fixation and the trophic landscape in the ocean. IMPORTANCE Marine plankton plays an important role in global carbon cycling and climate regulation. Phytoplankton and cyanobacteria fix CO2 to produce organic compounds using solar energy and mainly by the Calvin cycle, whereas autotrophic bacteria and archaea may fix CO2 by non-Calvin cycle carbon fixation pathways. How active individual lineages are in carbon fixation and mixotrophy, and what energy source bacteria may employ in non-Calvin carbon fixation, in a natural plankton assemblage are poorly understood and underexplored. Using metatranscriptomics, we studied carbon fixation in marine plankton with lineage resolution in tropical marginal shelf and slope areas. Based on the sequencing results, we characterized the carbon fixation potential of different lineages and assessed Calvin- and non-Calvin- carbon fixation activities and energy sources. Data revealed a high number of unigenes (4.4 million), lineage-dependent differential potentials of Calvin carbon fixation and responses to environmental conditions, major contributors of non-Calvin carbon fixation, and their potential energy source. C1 [Li, Hongfei; Yu, Liying; Li, Tangcheng; Li, Ling; Yuan, Huatao; Wang, Jingtian; Wang, Cong; Lin, Senjie] Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen, Fujian, Peoples R China. [Li, Hongfei] Zhejiang Ocean Univ, Natl Engn Res Ctr Marine Aquaculture, Zhoushan, Zhejiang, Peoples R China. [Li, Hongfei; Lin, Senjie] Univ Connecticut, Dept Marine Sci, Groton, CT 06340 USA. [Chen, Jianwei; Fan, Guangyi; Li, Denghui] BGI Res, Qingdao Key Lab Marine Genom, Qingdao, Shandong, Peoples R China. [Fan, Guangyi] BGI Res, State Key Lab Agr Genom, Shenzhen, Guangdong, Peoples R China. [Li, Denghui] Qingdao Innovat Ctr Seaweed Biotechnol, Qingdao, Shandong, Peoples R China. RP Lin, SJ (corresponding author), Xiamen Univ, State Key Lab Marine Environm Sci, Xiamen, Fujian, Peoples R China.; Lin, SJ (corresponding author), Univ Connecticut, Dept Marine Sci, Groton, CT 06340 USA. 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Field, Dawn Swift, Paul Newbold, Lindsay Oliver, Anna Smyth, Tim Somerfield, Paul J. Huse, Sue Joint, Ian TI The seasonal structure of microbial communities in the Western English Channel SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>Very few marine microbial communities are well characterized even with the weight of research effort presently devoted to it. Only a small proportion of this effort has been aimed at investigating temporal community structure. Here we present the first report of the application of high-throughput pyrosequencing to investigate intra-annual bacterial community structure. Microbial diversity was determined for 12 time points at the surface of the L4 sampling site in the Western English Channel. This was performed over 11 months during 2007. A total of 182 560 sequences from the V6 hyper-variable region of the small-subunit ribosomal RNA gene (16S rRNA) were obtained; there were between 11 327 and 17 339 reads per sample. Approximately 7000 genera were identified, with one in every 25 reads being attributed to a new genus; yet this level of sampling far from exhausted the total diversity present at any one time point. The total data set contained 17 673 unique sequences. Only 93 (0.5%) were found at all time points, yet these few lineages comprised 50% of the total reads sequenced. The most abundant phylum was Proteobacteria (50% of all sequenced reads), while the SAR11 clade comprised 21% of the ubiquitous reads and similar to 12% of the total sequenced reads. In contrast, 78% of all operational taxonomic units were only found at one time point and 67% were only found once, evidence of a large and transient rare assemblage. This time series shows evidence of seasonally structured community diversity. There is also evidence for seasonal succession, primarily reflecting changes among dominant taxa. 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Transporters with substrate specificities for nitrogen-and carbon-containing compounds (1.5% of the total spectra for each) were highly detected while no phosphate transporters were found, suggesting that carbon and nitrogen might be more limiting than phosphorus in the surface seawater. Viral proteins were assigned into three families: Myoviridae, Podoviridae and Siphoviridae, and the Myoviridae proteins were the most abundant. Among them, structure proteins were the most abundant viral proteins. This study indicated that the dissolved proteins of HMW-DOM presented compositional and biologically original homogeneity in the surface seawaters of the South China Sea, and bacteria and viruses dominated the dissolved protein pool. (C) 2013 Elsevier Ltd. All rights reserved. C1 [Dong, Hong-Po; Wang, Da-Zhi; Xie, Zhang-Xian; Dai, Min-Han; Hong, Hua-Sheng] Xiamen Univ, Coll Environm & Ecol, State Key Lab Marine Environm Sci, Xiamen, Peoples R China. 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Clone libraries of 16S rRNA gene fragments were constructed using environmental DNA collected from the OMZ (60 m and 200 m), the sea surface (10 m), and the deep oxycline (450 m). At the class level, the majority of sequences affiliated to the gamma- (53.7%) and alpha-Proteobacteria (19.7%), and to the Bacteroidetes (11.2%). A vertical partitioning of the bacterial communities was observed, with main differences between the suboxic OMZ and the more oxygenated surface and deep oxycline waters. At the surface, the microbial community was predominantly characterized by SAR86, Loktanella and unclassified Flavobacteriaceae, whereas the deeper layer was dominated by Sulfitobacter and unclassified Alteromonadaceae. In the OMZ, major constituents affiliated to the marine SAR11 clade and to thiotrophic gamma-symbionts (25% of all sequences), a group not commonly found in pelagic waters. Sequences affiliating to the phylum Chloroflexi, to the AGG47 and SAR202 clades, to the delta-Proteobacteria, to the Acidobacteria, and to the 'anammox group' of the Planctomycetes were found exclusively in the OMZ. The bacterial richness in the OMZ was higher than in the oxic surface and deeper oxycline, as revealed by rarefaction analysis and the Chao1 richness estimator (surface: 45 +/- 8, deeper oxycline: 76 +/- 26; OMZ(60m): 97 +/- 33, OMZ(200m): 109 +/- 31). OMZ bacterial diversity indices (Fisher's: similar to 30 +/- 5, Shannon's: similar to 3.31, inverse Simpson's: similar to 20) were similar to those found in other pelagic marine environments. Thus, our results indicate a distinct and diverse bacterial community within the OMZ, with presumably novel and yet uncultivated bacterial lineages. C1 Univ Concepcion, Lab Proc Oceanog & Clima, Dept Oceanog, Concepcion, Chile. Univ Concepcion, Lab Proc Oceanog & Clima, Ctr Invest Ocean & Pacifico Sud Oriental, Concepcion, Chile. 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Quiroga, Eduardo Alarcon, Guillermo Silva, Daniela Acuna, Carla Silva, Nelson TI Biogeochemical Responses and Seasonal Dynamics of the Benthic Boundary Layer Microbial Communities during the El Nino 2015 in an Eastern Boundary Upwelling System SO WATER DT Article AB The Eastern South Pacific coastal zone is characterized by seasonal and interannual variability, driven by upwelling and El Nino Southern Oscillation (ENSO), respectively. These oceanographical conditions influence microbial communities and their contribution to nutrient and greenhouse gases recycling, especially in bottom waters due to oxygenation. This article addresses the seasonal hydrographic and biogeochemical conditions in the water and sediments during El Nino 2015. Bottom water active microbial communities, including nitrifiers, were studied using amplicon sequencing of 16S rRNA (cDNA) and RT-qPCR, respectively. The results of the hydrographic analysis showed changes in the water column associated with the predominance of sub-Antarctic Waters characterized by warmed and low nutrients in the surface and more oxygenated conditions at the bottom in comparison with El Nino 2014. The organic matter quantity and quality decreased during fall and winter. The bottom water active microbial assemblages were dominated by archaea (Ca. Poseidoniales) and putative ammonia oxidizing archaea. Active bacteria affiliated to SAR11, Marinimicrobia and Nitrospina, and oxygen deficient realms (Desulfobacterales, SUP05 clade and anammox) suffered variations, possibly associated with oxygen and redox conditions in the benthic boundary layer. Nitrifying functional groups contributed significantly more during late fall and winter which was consistent with higher bottom water oxygenation. Relationships between apparent oxygen utilization nitrate and nitrous oxide in the water support the contribution of nitrification to this greenhouse gas distribution in the water. In general, our study suggests that seasonal oceanographic variability during an El Nino year influences the microbial community and thus remineralization potential, which supports the need to carry out longer time series to identify the relevance of seasonality under ENSO in Eastern Boundary Upwelling Systems (EBUS) areas. C1 [Molina, Veronica; Silva, Daniela; Acuna, Carla] Univ Playa Ancha, Observ Ecol Microbiana, Dept Biol, Fac Ciencias Nat & Exactas, Ave Leopoldo Carvallo 270, Valparaiso 2340000, Chile. [Molina, Veronica] Univ Playa Ancha, HUB Ambiental UPLA, Ave Leopoldo Carvallo 200, Valparaiso 2340000, Chile. [Cornejo-D'Ottone, Marcela; Quiroga, Eduardo; Alarcon, Guillermo; Silva, Nelson] Pontificia Univ Catolica Valparaiso, Escuela Ciencias Mar, Altamirano 1480, Valparaiso 2360007, Chile. [Cornejo-D'Ottone, Marcela; Quiroga, Eduardo; Alarcon, Guillermo; Silva, Nelson] Pontificia Univ Catolica Valparaiso, Inst Milenio Oceanog, Altamirano 1480, Valparaiso 2360007, Chile. [Soto, Eulogio H.] Univ Valparaiso, Fac Ciencias Mar & Recursos Nat, Ctr Observ Marino Estudios Riesgos Ambiente Coste, Casilla 5080, Vina Del Mar 2520000, Chile. [Soto, Eulogio H.] Univ Valparaiso, Lab Bentos, Escuela Biol Marina, Vina Del Mar 2520000, Chile. RP Molina, V (corresponding author), Univ Playa Ancha, Observ Ecol Microbiana, Dept Biol, Fac Ciencias Nat & Exactas, Ave Leopoldo Carvallo 270, Valparaiso 2340000, Chile.; Molina, V (corresponding author), Univ Playa Ancha, HUB Ambiental UPLA, Ave Leopoldo Carvallo 200, Valparaiso 2340000, Chile.; Cornejo-D'Ottone, M (corresponding author), Pontificia Univ Catolica Valparaiso, Escuela Ciencias Mar, Altamirano 1480, Valparaiso 2360007, Chile.; Cornejo-D'Ottone, M (corresponding author), Pontificia Univ Catolica Valparaiso, Inst Milenio Oceanog, Altamirano 1480, Valparaiso 2360007, Chile. 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This effective N-2-yielding section encloses a suspended small-particle layer, inferred from particle backscattering (b(bp)) measurements. It is thus hypothesized that this layer (hereafter, the b(bp)-layer) is linked to microbial communities involved in N-2 yielding such as nitrate-reducing SAR11 as well as sulfur-oxidizing, anammox, and denitrifying bacteria - a hypothesis yet to be evaluated. Here, data collected by three BGC-Argo floats deployed in the Black Sea are used to investigate the origin of this bbp-layer. To this end, we evaluate how the key drivers of N-2-yielding bacteria dynamics impact the vertical distribution of b(bp) and the thickness of the b(bp)-layer. In conjunction with published data on N-2 excess, our results suggest that the b(bp)-layer is at least partially composed of the bacteria driving N-2 yielding for three main reasons: (1) strong correlations are recorded between bbp and nitrate; (2) the top location of the b(bp)-layer is driven by the ventilation of oxygen-rich subsurface waters, while its thickness is modulated by the amount of nitrate available to produce N-2; and (3) the maxima of both b(bp) and N-2 excess coincide at the same isopycnals where bacteria involved in N-2 yielding coexist. We thus advance that b(bp) and O-2 can be exploited as a combined proxy to delineate the N-2-yielding section of the Black Sea. This proxy can potentially contribute to refining delineation of the effective N-2-yielding section of oxygen-deficient zones via data from the growing BGC-Argo float network. C1 [Rasse, Rafael; Claustre, Herve; Poteau, Antoine] Sorbonne Univ, F-06230 Villefranche Sur Mer, France. 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TI Transient Changes in Bacterioplankton Communities Induced by the Submarine Volcanic Eruption of El Hierro (Canary Islands) SO PLOS ONE DT Article AB The submarine volcanic eruption occurring near El Hierro (Canary Islands) in October 2011 provided a unique opportunity to determine the effects of such events on the microbial populations of the surrounding waters. The birth of a new underwater volcano produced a large plume of vent material detectable from space that led to abrupt changes in the physicalchemical properties of the water column. We combined flow cytometry and 454-pyrosequencing of 16S rRNA gene amplicons (V1-V3 regions for Bacteria and V3-V5 for Archaea) to monitor the area around the volcano through the eruptive and post-eruptive phases (November 2011 to April 2012). Flow cytometric analyses revealed higher abundance and relative activity (expressed as a percentage of high-nucleic acid content cells) of heterotrophic prokaryotes during the eruptive process as compared to post-eruptive stages. Changes observed in populations detectable by flow cytometry were more evident at depths closer to the volcano (similar to 70-200 m), coinciding also with oxygen depletion. Alpha-diversity analyses revealed that species richness (Chao1 index) decreased during the eruptive phase; however, no dramatic changes in community composition were observed. The most abundant taxa during the eruptive phase were similar to those in the post-eruptive stages and to those typically prevalent in oceanic bacterioplankton communities (i.e. the alphaproteobacterial SAR11 group, the Flavobacteriia class of the Bacteroidetes and certain groups of Gammaproteobacteria). Yet, although at low abundance, we also detected the presence of taxa not typically found in bacterioplankton communities such as the Epsilonproteobacteria and members of the candidate division ZB3, particularly during the eruptive stage. These groups are often associated with deep-sea hydrothermal vents or sulfur-rich springs. Both cytometric and sequence analyses showed that once the eruption ceased, evidences of the volcano-induced changes were no longer observed. C1 [Ferrera, Isabel; Gasol, Josep M.] CSIC, Inst Ciencias Mar, Dept Biol Marina & Oceanog, Barcelona, Spain. [Aristegui, Javier; Montero, Maria F.] Univ Las Palmas Gran Canaria, Inst Oceanog & Cambio Global, Las Palmas Gran Canaria, Spain. [Gonzalez, Jose M.] Univ La Laguna, Dept Microbiol, E-38207 San Cristobal la Laguna, Spain. [Fraile-Nuez, Eugenio] Ctr Oceanog Canarias, Inst Espanol Oceanog, Santa Cruz De Tenerife, Spain. 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Sun, Shulei Mou, Xiaozhen Moran, Mary Ann TI Transporter genes expressed by coastal bacterioplankton in response to dissolved organic carbon SO ENVIRONMENTAL MICROBIOLOGY DT Article AB P>Coastal ocean bacterioplankton control the flow of dissolved organic carbon (DOC) from terrestrial and oceanic sources into the marine food web, and regulate the release of inorganic carbon to atmospheric and offshore reservoirs. While the fate of the chemically complex coastal DOC reservoir has long been recognized as a critical feature of the global carbon budget, it has been problematic to identify both the compounds that serve as major conduits for carbon flux and the roles of individual bacterioplankton taxa in mediating that flux. Here we analyse random libraries of expressed genes from a coastal bacterial community to identify sequences representing DOC-transporting proteins. Predicted substrates of expressed transporter genes indicated that carboxylic acids, compatible solutes, polyamines and lipids may be key components of the biologically labile DOC pool in coastal waters, in addition to canonical bacterial substrates such as amino acids, oligopeptides and carbohydrates. Half of the expressed DOC transporter sequences in this coastal ocean appeared to originate from just eight taxa: Roseobacter, SAR11, Flavobacteriales and five orders of gamma-Proteobacteria. While all major taxa expressed transporter genes for some DOC components (e.g. amino acids), there were indications of specialization within the bacterioplankton community for others (e.g. carbohydrates, carboxylic acids and polyamines). Experimental manipulations of the natural DOC pool that increased the concentration of phytoplankton- or vascular plant-derived compounds invoked a readily measured response in bacterial transporter gene expression. This highly resolved view of the potential for carbon flux into heterotrophic bacterioplankton cells identifies possible bioreactive components of the coastal DOC pool and highlights differing ecological roles in carbon turnover for the resident bacterial taxa. C1 [Poretsky, Rachel S.; Sun, Shulei; Mou, Xiaozhen; Moran, Mary Ann] Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. RP Moran, MA (corresponding author), Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. 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Jankowiak, Jennifer Koch, Florian Gobler, Christopher J. TI Prokaryotic and eukaryotic microbiomes associated with blooms of the ichthyotoxic dinoflagellate Cochlodinium (Margalefidinium) polykrikoides in New York, USA, estuaries SO PLOS ONE DT Article AB While harmful algal blooms caused by the ichthyotoxic dinoflagellate, Cochlodinium (Margalefidinium) polykrikoides, are allelopathic and may have unique associations with bacteria, a comprehensive assessment of the planktonic communities associated with these blooms has been lacking. Here, we used high-throughput amplicon sequencing to assess size fractionated (0.2 and 5 mu m) bacterial (16S) and phytoplankton assemblages (18S) associated with blooms of C. polykrikoides during recurrent blooms in NY, USA. Over a three-year period, samples were collected inside ('patch') and outside ('non-patch') dense accumulations of C. polykrikoides to assess the microbiome associated with these blooms. Eukaryotic plankton communities of blooms had significantly lower diversity than non-bloom samples, and non-bloom samples hosted 30 eukaryotic operational taxonomic units (OTUs) not found within blooms, suggesting they may have been allelopathically excluded from blooms. Differential abundance analyses revealed that C. polykrikoides blooms were significantly enriched in dinoflagellates (p<0.001) and the experimental enrichment of C. polykrikoides led to a significant increase in the relative abundance of eight genera of dinoflagellates but a significant decline in other eukaryotic plankton. Amoebophrya co-dominated both within- and near- C. polykrikoides blooms and was more abundant in bloom patches. The core bacterial microbiome of the >0.2 mu m fraction of blooms was dominated by an uncultured bacterium from the SAR11 clade, while the >5 mu m size fraction was co-dominated by an uncultured bacterium from Rhodobacteraceae and Coraliomargarita. Two bacterial lineages within the >0.2 mu m fraction, as well as the Gammaproteobacterium, Halioglobus, from the >5 mu m fraction were unique to the microbiome of blooms, while there were 154 bacterial OTUs only found in non-bloom waters. Collectively, these findings reveal the unique composition and potential function of eukaryotic and prokaryotic communities associated with C. polykrikoides blooms. C1 [Hattenrath-Lehmann, Theresa K.; Jankowiak, Jennifer; Koch, Florian; Gobler, Christopher J.] SUNY Stony Brook, Sch Marine & Atmospher Sci, Southampton, NY 11968 USA. [Jankowiak, Jennifer] New York State Dept Hlth, Wadsworth Ctr, Empire State Plaza, Albany, NY USA. [Koch, Florian] Helmholtz Zentrum Polar & Meeresforsch, Alfred Wegener Inst, Marine Biogeosci, Bremerhaven, Germany. RP Gobler, CJ (corresponding author), SUNY Stony Brook, Sch Marine & Atmospher Sci, Southampton, NY 11968 USA. 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In last decades, the study of marine microbial ecology is increasingly reliant on molecular-based methods, and as a result many of the research projects conducted in such courses require sequencing that is often not available on site and may take more time than a typical course allows. In this work, we describe a protocol combining molecular and functional methods for analyzing proteorhodopsins (PRs), with visible results in only 4-5 days that do not rely on sequencing. PRs were discovered in oceanic surface waters two decades ago, and have since been observed in different marine environments and diverse taxa, including the abundant alphaproteobacterial SAR11 group. PR subgroups are currently known to absorb green and blue light, and their distribution was previously explained by prevailing light conditions - green pigments at the surface and blue pigments in deeper waters, as blue light travels deeper in the water column. 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The picoplankton communities reached their maximum abundance in the spring euphotic zone when the maximum value of the chlorophyll a in response to deep winter convection was recorded. The communities were dominated by Bacteria, while Archaea were a minor constituent. A seasonality of bacterial richness and diversity was observed, with minimum values occurring during the winter convection and spring postconvection periods and maximum values occurring under summer stratified conditions. The SAR11 clade was the main constituent of the bacterial communities and reached the maximum abundance in the euphotic zone in spring after the convection episode. Cyanobacteria were the second most abundant group, and their abundance strongly depended on the convection event, when minimal cyanobacterial abundance was observed. In spring and autumn, the euphotic zone was characterized by Bacteroidetes and Gammaproteobacteria. 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In this study, bacterial communities and diversity along the shores of the Upper Gulf of Thailand were first characterized. The association between bacterial communities and types of land use was also evaluated. Results The bacterial communities and diversity of seawater in the Upper Gulf of Thailand, with regard to types of land use, were first revealed by using Illumina next-generation sequencing. A total of 4953 OTUs were observed from all samples in which 554 OTUs were common. The bacterial communities in sampling sites were significantly different from each other. The run-off water from three types of land use significantly affected the community richness and diversity of marine bacteria. Aquaculture sites contained the highest levels of community richness and diversity, followed by mangrove forests and tourist sites. Seawater physicochemical parameters including salinity, turbidity, TSS, total N, and BOD5, were significantly different when grouped by land use. The bacterial communities were mainly determined by salinity, total N, and total P. The species richness estimators and OTUs were positively correlated with turbidity. The top ten most abundant phyla and genera as well as the distribution of bacterial classes were characterized. The Proteobacteria constituted the largest proportions in all sampling sites, ranging between 67.31 and 78.80%. The numbers of the Marinobacterium, Neptuniibacter, Synechococcus, Candidatus Thiobios, hgcI clade (Actinobacteria), and Candidatus Pelagibacter were significantly different when grouped by land use. Conclusions Type of land use significantly affected bacterial communities and diversity along the Upper Gulf of Thailand. Turbidity was the most influential parameter affecting the variation in bacterial community composition. Salinity, total N, and P were the ones of the important factors that shaped the bacterial communities. In addition, the variations of bacterial communities from site-to-site were greater than within-site. The Proteobacteria, Bacteroidetes, Actinobacteria, Cyanobacteria, Verrucomicrobia, Euryarchaeota, Planctomycetes, Firmicutes, Deep Sea DHVEG-6, and Marinimicrobia were the most and common phyla distributed across the Upper Gulf of Thailand. C1 [Nimnoi, Pongrawee] Kasetsart Univ, Dept Microbiol, Fac Liberal Arts & Sci, Nakhon Pathom, Thailand. [Pongsilp, Neelawan] Silpakorn Univ, Dept Microbiol, Fac Sci, Nakhon Pathom, Thailand. RP Pongsilp, N (corresponding author), Silpakorn Univ, Dept Microbiol, Fac Sci, Nakhon Pathom, Thailand. 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Flavobacterium, Candidatus_Aquiluna, SAR11 clade, Polaribacter, Glaciecola, and Nonlabens were the dominant genera observed in the amplicon analysis. Whereas Actimicrobium, Rhodoglobus, Flavobacterium, and Pseudomonas were dominated in the culturable fraction. Our results also demonstrated that salinity, chlorophyll a, and dissolved organic carbon were the significant environmental variables controlling the prokaryotic community distribution in melt ponds. A significant community shift was observed in melt ponds when the salinity changed with the progression of melting and deepening of ponds. Different communities were found to be dominant in melt ponds with different salinity ranges. It was also observed that melt pond prokaryotic communities significantly differed from the surface ocean microbial community. 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Using 16S metabarcoding, we analyzed samples of prokaryotic plankton communities in the Barents Sea and found two types of communities. The origin of these communities is discussed in terms of biogeography.Abstract The Barents Sea is one of the most rapidly changing Arctic regions, with an unprecedented sea ice decline and increase in water temperature and salinity. We have studied the diversity of prokaryotic communities using 16S metabarcoding in the western and northeastern parts of the Barents Sea along the Kola Section and the section from Novaya Zemlya to Franz Joseph Land. The hypothesis-independent clustering method revealed the existence of two distinct types of communities. The most common prokaryotic taxa were shared between two types of communities, but their relative abundance was different. It was found that the geographic location of the sampling sites explained more than 30% of the difference between communities, while no statistically significant correlation between environmental parameters and community composition was found. The representatives of the Psychrobacter, Sulfitobacter and Polaribacter genera were dominant in samples from both types of communities. The first type of community was also dominated by members of Halomonas, Pseudoalteromonas, Planococcaceae and an unclassified representative of the Alteromonadaceae family. The second type of community also had a significant proportion of Nitrincolaceae, SAR92, SAR11 Clade I, NS9, Cryomorphaceae and SUP05 representatives. The origin of these communities can be explained by the influence of environmental factors or by the different origins of water masses. This research highlights the importance of studying biogeographic patterns in the Barents Sea in comparison with those in the North Atlantic and Arctic Ocean prokaryote communities. C1 [Namsaraev, Zorigto; Kozlova, Aleksandra; Krylova, Anastasia; Izotova, Anna; Melnikova, Anna; Patrushev, Maksim; Toshchakov, Stepan] Kurchatov Inst, Natl Res Ctr, Kurchatov Ctr Genome Res, Moscow 123182, Russia. [Namsaraev, Zorigto; Kuzmin, Denis] Moscow Inst Phys & Technol, Dolgoprudnyi 141701, Russia. [Tuzov, Fedor] Lomonosov Moscow State Univ, Fac Geog, Dept Oceanol, Moscow 119991, Russia. [Tuzov, Fedor] All Russian Res Inst Civil Def & Emergencies, Moscow 121352, Russia. [Bezgreshnov, Andrei] Arctic & Antarctic Res Inst, St Petersburg 199397, Russia. [Trofimova, Anna] Northern Arctic Fed Univ, Higher Sch Nat Sci & Technol, Dept Geog & Hydrometeorol, Arkhangelsk 163002, Russia. 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Our results showed that viral regulation occurring at the strain level can affect the bacterial community structure at higher phylogenetic levels. In addition, some less abundant groups, such as Vibrio and Alteromonas, might actually win the competition for resources, providing viral control is removed. Our results also showed that the population dynamics of certain bacterial groups was controlled by viruses, and the strength of such control was modulated by the concentration of DOM. We suggest that the composition of the bacterial community in aquatic environments results from the interactive regulation by DOM and viruses among other regulators. C1 [Liu, Hongbin; Tan, Shangjin; Guo, Wang; Xia, Xiaomin; Cheung, Shun Yan] Hong Kong Univ Sci & Technol, Div Life Sci, Kowloon, Hong Kong, Peoples R China. [Xu, Jie] Chinese Acad Sci, South China Sea Inst Oceanol, State Key Lab Trop Oceanog, Guangzhou, Guangdong, Peoples R China. RP Liu, HB (corresponding author), Hong Kong Univ Sci & Technol, Div Life Sci, Kowloon, Hong Kong, Peoples R China. 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Vergin, Kevin Bolanos, Luis M. Giovannoni, Stephen J. Parsons, Rachel Opalk, Keri Halewood, Elisa Hansell, Dennis A. Johnson, Rod Curry, Ruth Carlson, Craig A. TI Linkages Among Dissolved Organic Matter Export, Dissolved Metabolites, and Associated Microbial Community Structure Response in the Northwestern Sargasso Sea on a Seasonal Scale SO FRONTIERS IN MICROBIOLOGY DT Article AB Deep convective mixing of dissolved and suspended organic matter from the surface to depth can represent an important export pathway of the biological carbon pump. The seasonally oligotrophic Sargasso Sea experiences annual winter convective mixing to as deep as 300 m, providing a unique model system to examine dissolved organic matter (DOM) export and its subsequent compositional transformation by microbial oxidation. We analyzed biogeochemical and microbial parameters collected from the northwestern Sargasso Sea, including bulk dissolved organic carbon (DOC), total dissolved amino acids (TDAA), dissolved metabolites, bacterial abundance and production, and bacterial community structure, to assess the fate and compositional transformation of DOM by microbes on a seasonal time-scale in 2016-2017. DOM dynamics at the Bermuda Atlantic Time-series Study site followed a general annual trend of DOC accumulation in the surface during stratified periods followed by downward flux during winter convective mixing. Changes in the amino acid concentrations and compositions provide useful indices of diagenetic alteration of DOM. TDAA concentrations and degradation indices increased in the mesopelagic zone during mixing, indicating the export of a relatively less diagenetically altered (i.e., more labile) DOM. During periods of deep mixing, a unique subset of dissolved metabolites, such as amino acids, vitamins, and benzoic acids, was produced or lost. DOM export and compositional change were accompanied by mesopelagic bacterial growth and response of specific bacterial lineages in the SAR11, SAR202, and SAR86 clades, Acidimicrobiales, and Flavobacteria, during and shortly following deep mixing. Complementary DOM biogeochemistry and microbial measurements revealed seasonal changes in DOM composition and diagenetic state, highlighting microbial alteration of the quantity and quality of DOM in the ocean. C1 [Liu, Shuting; Opalk, Keri; Halewood, Elisa; Carlson, Craig A.] Univ Calif, Inst Marine Sci, Dept Ecol, Evolut & Marine Biol, Santa Barbara, CA USA. [Longnecker, Krista; Kujawinski, Elizabeth B.] Dept Marine Chem & Geochem, Woods Hole OceanographicInstitut, Woods Hole, MA USA. [Vergin, Kevin] Microbial DNA Analyt, Phoenix, OR USA. 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The smallest globin-bearing genomes are the streamlined genomes (similar to 1.3 Mbp) of the SAR11 clade of alphaproteobacteria and the slightly larger (ca.1.7 Mbp) genomes of Aquificae. The smallest genome with members of all three families is the 23 Mbp genome of the extremophile Methylacidiphilum infernorum (Verrumicrobia). Of the 147 possible combinations of the eight globin subfamilies, only 83 are observed. Although binary combinations are infrequent and ternary combinations are rare, the FHb + TrHb2 combination is the most commonly observed. Of the possible functions of bacterial globins we discuss the two principal - ones nitric oxide detoxification via the NO dioxygenase or denitrosylase activities and the sensing of oxygen concentration in the environmental niche. In only few cases has a physiological role been demonstrated in vivo. This article is part of a Special Issue entitled: Oxygen Binding and Sensing Proteins. (c) 2013 Elsevier B.V. All rights reserved. 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Studies to date suggested a combination of deterministic (where biotic/abiotic factors act on fitness differences amongst taxa) and stochastic (where dispersal plays a larger factor in community assembly) processes are responsible for structuring biotic communities, but there is no consensus on the relative roles these processes play, and data is lacking for lakes. In the present study, we sampled different biotic components in 34 lakes located on the South Island of New Zealand. To obtain a holistic view of assembly processes in lakes we used metabarcoding to investigate bacteria in the sediment and surface waters, and eukaryotes in the sediment and two different size fractions of the water column. Physicochemical parameters were collected in parallel. Results showed that deterministic processes dominated the assembly of lake communities although the relative importance of variable and homogeneous selection differed among the biotic components. 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Damashek, Julian Eloe-Fadrosh, Emiley A. Francis, Christopher A. TI In-depth Spatiotemporal Characterization of Planktonic Archaeal and Bacterial Communities in North and South San Francisco Bay SO MICROBIAL ECOLOGY DT Article AB Despite being the largest estuary on the west coast of North America, no in-depth survey of microbial communities in San Francisco Bay (SFB) waters currently exists. In this study, we analyze bacterioplankton and archaeoplankton communities at several taxonomic levels and spatial extents (i.e., North versus South Bay) to reveal patterns in alpha and beta diversity. We assess communities using high-throughput sequencing of the 16S rRNA gene in 177 water column samples collected along a 150-km transect over a 2-year monthly time-series. In North Bay, the microbial community is strongly structured by spatial salinity changes while in South Bay seasonal variations dominate community dynamics. Along the steep salinity gradient in North Bay, we find that operational taxonomic units (OTUs; 97% identity) have higher site specificity than at coarser taxonomic levels and turnover ("species" replacement) is high, revealing a distinct brackish community (in oligo-, meso-, and polyhaline samples) from fresh and marine end-members. At coarser taxonomic levels (e.g., phylum, class), taxa are broadly distributed across salinity zones (i.e., present/abundant in a large number of samples) and brackish communities appear to be a mix of fresh and marine communities. We also observe variations in brackish communities between samples with similar salinities, likely related to differences in water residence times between North and South Bay. Throughout SFB, suspended particulate matter is positively correlated with richness and influences changes in beta diversity. Within several abundant groups, including the SAR11 clade (comprising up to 30% of reads in a sample), OTUs appear to be specialized to a specific salinity range. Some other organisms also showed pronounced seasonal abundance, including Synechococcus, Ca. Actinomarina, and Nitrosopumilus-like OTUs. Overall, this study represents the first in-depth spatiotemporal survey of SFB microbial communities and provides insight into how planktonic microorganisms have specialized to different niches along the salinity gradient. C1 [Rasmussen, Anna N.; Damashek, Julian; Francis, Christopher A.] Stanford Univ, Dept Earth Syst Sci, 473 Via Ortega,Y2E2 Bldg Rm 140, Stanford, CA 94305 USA. [Damashek, Julian] Utica Coll, Dept Biol, Utica, NY 13502 USA. [Eloe-Fadrosh, Emiley A.] Lawrence Berkeley Natl Lab, Joint Genome Inst, Dept Energy, Berkeley, CA 94720 USA. RP Francis, CA (corresponding author), Stanford Univ, Dept Earth Syst Sci, 473 Via Ortega,Y2E2 Bldg Rm 140, Stanford, CA 94305 USA. 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TI Spatial patterns of bacterial and archaeal communities along the Romanche Fracture Zone (tropical Atlantic) SO FEMS MICROBIOLOGY ECOLOGY DT Article AB The composition of prokaryotic communities was determined in the meso- and bathypelagic waters funneled through the Romanche Fracture Zone (RFZ, 2 degrees 7'S, 31 degrees 79'W to 0 degrees 6'N, 14 degrees 33'W) in the tropical Atlantic. Distinct water masses were identified based on their physical and chemical characteristics. The bacterial and archaeal communities were depth-stratified with a total of 116 and 25 operational taxonomic units (OTUs), respectively, distributed among the distinct water masses as revealed by terminal restriction fragment length polymorphism, and cloning and sequencing. The relative abundance of Thaumarchaeota, determined by catalyzed reporter deposition-fluorescence in situ hybridization, was significantly higher in deeper layers (Antarctic Bottom Water, AABW, > 4000 m depth), contributing up to 31% to the total prokaryotic community, than in the mesopelagic and lower euphotic layer. Although the contribution of SAR11 to bacterial abundance did not increase with depth, SAR202, SAR324, SAR406 and Alteromonas did increase with depth. Terminal restriction fragment length polymorphism analysis revealed successional changes in the bacterial and archaeal community composition of the North Atlantic Deep Water (NADW) with a passage time through the RFZ of c. 4 months but not in the under- and overlying water masses. Our results indicate that specific water masses harbor distinct bacterial and archaeal communities and that the prokaryotic community of the NADW undergoes successional changes in this conduit between the western and eastern Atlantic basin. 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Casotti, Raffaella TI Group-specific effects on coastal bacterioplankton of polyunsaturated aldehydes produced by diatoms SO AQUATIC MICROBIAL ECOLOGY DT Article AB Polyunsaturated aldehydes (PUAs), produced as secondary metabolites by diatoms, have been shown to induce toxic effects on a variety of organisms, including copepods and phyto- and bacterioplankton. However, the nature of and the players in this interaction remain poorly understood. We tested the effect of 3 PUAs commonly produced by marine diatoms-2E, 4E/Z-heptadienal (HEPTA), 2E, 4E/Z-octadienal (OCTA), 2E, 4E/Z-decadienal (DECA) and a mix of HEPTA and OCTA (MIX)-on a natural bacterial community from a coastal area of the NW Mediterranean Sea (Blanes Bay, Spain). Little effect on total or relative cell abundance or bulk bacterial production was observed after 6 or 24 h exposure to 7.5 nM of the 3 different PUAs for the different bacterial phylogenetic groups (Gammaproteobacteria, Bacteroidetes, Rhodobacteraceae and SAR11), assessed by catalysed reporter deposition (CARD)-fluorescence in situ hybridisation (FISH). Metabolic activity, i.e. single-cell activity as determined by microautoradiography combined with CARD-FISH (MAR-CARD-FISH), was least affected by the addition of single PUAs in Gammaproteobacteria, markedly in Bacteroidetes and most markedly in Rhodobacteraceae, leading to a decrease in Rhodobacteraceae abundance by 21% (by 38% of the active cells assessed by leucine uptake) compared to the control. Bacteroidetes, although markedly affected in single-cell activity, were the most abundant group (54% of total cell counts). The addition of a mixture of OCTA and HEPTA produced a more pronounced decrease in the metabolic activity of all groups than the incubation with the single PUAs, suggesting a synergistic effect. Our results demonstrate that PUAs have a differential effect on the single-cell activity of distinct bacterial groups in natural communities. PUAs may therefore play an important role in shaping bacterial community composition by conferring a competitive advantage to PUA-resistant groups, allowing them to preferentially use the organic matter released by diatoms. C1 [Balestra, Cecilia; Casotti, Raffaella] Stn Zool A Dohrn, I-80121 Naples, Italy. [Alonso-Saez, Laura; Gasol, Josep M.] CSIC, Inst Ciencies Mar, E-08003 Barcelona, Catalonia, Spain. RP Casotti, R (corresponding author), Stn Zool A Dohrn, Villa Comunale, I-80121 Naples, Italy. 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These pronounced changes in microbiome structure were also apparent when we examined the core microbiome of different tissue types. Although the core microbiome associated with healthy leaves included three operational taxonomic units (OTUs) classified as Burkholderia, Cryomorphaceae and the SAR11 clade, a single core OTU from the Arenicella was found within adjacent tissues. Burkholderia are diazotrophic microorganisms and may play an important role in seagrass nitrogen acquisition. In contrast, some members of the Arenicella have been implicated in necrotic disease in other benthic animals. Moreover, microbiome structure was maintained across sites within healthy tissues, but not within SWD lesions or the tissues immediately adjacent to lesions. Predicted functional profiles revealed increased photoautotrophic functions in SWD tissues relative to healthy leaves, but no increase in pathogenicity or virulence. 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After analyzing 1.6 million sequences from 16S rDNA and rRNA (cDNA) libraries from two euphotic depths from a representative site in the Pacific Ocean, we show that although there is an overall positive relationship between the abundances (rDNAs) and activities (rRNAs) among populations of the bacterial community, for some populations these measures are uncoupled. Different ecological strategies are exemplified by the two numerically dominant clades at this site: the cyanobacterium Prochlorococcus is abundant but disproportionately more active, while the heterotrophic SAR11 is abundant but less active. Other rare populations, such as Alteromonas, have high specific activities in spite of their low abundances, suggesting intense population regulation. More detailed analyses using a complementary quantitative PCR (qPCR)-based approach of measuring relative specific activity for Prochlorococcus populations in the Pacific and Atlantic Oceans also show that specific activity, but not abundance, reflects the key drivers of light and nutrients in this system; our results also suggest substantial top-down regulation (e.g., grazing, viruses, or organismal interactions) or transport (e.g., mixing, immigration, or emigration) of these populations. Thus, we show here that abundance and specific activity can be uncoupled in open ocean systems and that describing both is critical to characterizing microbial communities and predicting marine ecosystem functioning and responses to change. C1 [Hunt, Dana E.; Lin, Yajuan; Izzo, Lisa K.; Johnson, Zackary I.] Duke Univ, Marine Lab, Nicholas Sch Environm, Beaufort, NC 28516 USA. [Church, Matthew J.; Karl, David M.] Univ Hawaii, Dept Oceanog, Honolulu, HI 96822 USA. 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Catalyzed reporter deposition-fluorescence in situ hybridization showed that Alphaproteobacteria (average relative abundance 37%, average absolute abundance 3.7 x 10(5) cells ml(-1)) including SAR11 [30% / (3 x 10(5))], Gammaproteobacteria [14% / (1.2 x 10(5))] and Bacteroidetes [12% / (1.3 x 10(5))] globally dominated the bacterioplankton. The SAR86 clade [4.6% / (4.1 x 10(4))] and Actinobacteria [4.5% / (4 x 10(4))] were detected ubiquitously, whereas Archaea were scarce [0.6% / (4.2 x 10(3))]. The Roseobacter clade [averaging 3.8% / (3.5 x 10(4))], Pseudoalteromonas [2.6% / (2.1 x 10(4))] and Vibrio [1.5% / (1.3 x 10(4))] showed cosmopolitan occurrence. Principal component analysis revealed a latitudinal pattern in bacterial abundances by clustering samples according to lower and higher latitudes. This was related to significantly different relative abundances of Bacteroidetes (peaking at higher latitudes) and of unclassified Bacteria and Vibrio (both peaking at lower latitudes) between warmer and colder oceans. Relative abundances of Alphaproteobacteria (peaking at subtropical) and Gammaproteobacteria (polar stations) varied between major oceanic biomes, as did absolute abundances of Roseobacter (peaking at temperate and polar stations). For almost all groups, absolute abundances were positively correlated with nutrient concentrations in warmer oceans and negatively correlated with oxygen saturation in colder oceans. On a global scale, Roseobacter and SAR86 were correlated with chlorophyll a. Linkages of environmental parameters with relative abundances were more complex, with e. g. Bacteroidetes being associated with chlorophyll a. 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Olsen, Gary J. TI Characterizing the Native Codon Usages of a Genome: An Axis Projection Approach SO MOLECULAR BIOLOGY AND EVOLUTION DT Article AB Codon usage can provide insights into the nature of the genes in a genome. Genes that are "native" to a genome (have not been recently acquired by horizontal transfer) range in codon usage from a low-bias "typical" usage to a more biased "high-expression" usage characteristic of genes encoding abundant proteins. Genes that differ from these native codon usages are candidates for foreign genes that have been recently acquired by horizontal gene transfer. In this study, we present a method for characterizing the codon usages of native genes-both typical and highly expressed-within a genome. Each gene is evaluated relative to a half line (or axis) in a 59D space of codon usage. 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Community structure did not differ greatly around the Cape Verde Islands between sampling periods but varied substantially in the open ocean, suggesting different environmental perturbations favour specific bacterial groups. Wind speed varied significantly between years, with moderate to strong breeze in winter 2008 and gales in winter 2006 (8.9 +/- 0.2 ms(-1) and 16.0 +/- 0.4 ms(-1), respectively). Enhanced wind-driven turbulence was associated with domination by the SAR11 clade of Alphaproteobacteria, which were present at 2.4-fold in the abundance of Prochlorococcus (41.8 +/- 1.6% cells, compared to 17.7 +/- 7.1%). Conversely, the calmer conditions of 2008 seemed to favour Prochlorococcus (40.0 +/- 1.2% cells). Prochlorococcus high-light adapted clade HLI were only numerous during wind-driven turbulence, whereas oligotrophic-adapted clade HLII dominated under calm conditions. Bacteroidetes were most prominent in turbulent conditions (9.5 +/- 1.3% cells as opposed to 4.7 +/- 0.3%), as were Synechococcus. In 2008, a considerable dust deposition event occurred in the region, which may have led to the substantial Gammaproteobacteria population (22.5 +/- 4.0% cells compared to 4.6 +/- 0.6% in 2006). Wind-driven turbulence may have a significant impact on microbial community structure in the surface ocean. Therefore, community change following dust storm events may be linked to associated wind in addition to dust-derived nutrients. C1 [Hill, Polly G.; Purdie, Duncan A.] Univ Southampton, Natl Oceanog Ctr, Southampton SO14 3ZH, Hants, England. [Heywood, Jane L.] Univ Bremen, Fac Biol Chem, D-28359 Bremen, Germany. [Fuchs, Bernhard M.] Max Planck Inst Microbiol, D-28359 Bremen, Germany. RP Hill, PG (corresponding author), Univ Southampton, Natl Oceanog Ctr, European Way, Southampton SO14 3ZH, Hants, England. 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It revealed the spatiotemporal dynamics of the unique viral assemblages and auxiliary metabolic genes (AMGs) following the variation and degradation of Ulva prolifera. These findings demonstrate a tight coupling between viral assemblages, and prokaryotic and eukaryotic abundances were influenced by the green tide. The world's largest macroalgal green tide, caused by Ulva prolifera, has resulted in serious consequences for coastal waters of the Yellow Sea, China. Although viruses are considered to be one of the key factors in controlling microalgal bloom demise, understanding of the relationship between viral communities and the macroalgal green tide is still poor. Here, a Qingdao coastal virome (QDCV) time-series data set was constructed based on the metagenomic analysis of 17 DNA viromes along three coastal stations of the Yellow Sea, covering different stages of the green tide from Julian days 165 to 271. A total of 40,076 viral contigs were detected and clustered into 28,058 viral operational taxonomic units (vOTUs). About 84% of the vOTUs could not be classified, and 62% separated from vOTUs in other ecosystems. Green tides significantly influenced the spatiotemporal dynamics of the viral community structure, diversity, and potential functions. For the classified vOTUs, the relative abundance of Pelagibacter phages declined with the arrival of the bloom and rebounded after the bloom, while Synechococcus and Roseobacter phages increased, although with a time lag from the peak of their hosts. More than 80% of the vOTUs reached peaks in abundance at different specific stages, and the viral peaks were correlated with specific hosts at different stages of the green tide. Most of the viral auxiliary metabolic genes (AMGs) were associated with carbon and sulfur metabolism and showed spatiotemporal dynamics relating to the degradation of the large amount of organic matter released by the green tide.IMPORTANCE To the best of our knowledge, this study is the first to investigate the responses of viruses to the world's largest macroalgal green tide. It revealed the spatiotemporal dynamics of the unique viral assemblages and auxiliary metabolic genes (AMGs) following the variation and degradation of Ulva prolifera. These findings demonstrate a tight coupling between viral assemblages, and prokaryotic and eukaryotic abundances were influenced by the green tide. C1 [Han, Meiaoxue; Sun, Jianhua; Yang, Qingwei; Liang, Yantao; Jiang, Yong; Gao, Chen; Gu, Chengxiang; Liu, Qian; Chen, Xuechao; Liu, Gang; Shao, Hongbing; Guo, Cui; He, Hui; Wang, Hualong; McMinn, Andrew; Wang, Min] Ocean Univ China, Inst Evolut & Marine Biodivers, Coll Marine Life Sci, Key Lab Polar Oceanog & Global Ocean Change, Qingdao, Peoples R China. [Liang, Yantao; Shao, Hongbing; Guo, Cui; He, Hui; Wang, Hualong; Sung, Yeong Yik; Mok, Wen Jye; Wong, Li Lian; Wang, Min] Univ Malaysia Terengganu Ocean Univ China Joint Ct, Qingdao, Peoples R China. [Sung, Yeong Yik; Mok, Wen Jye; Wong, Li Lian] Univ Malaysia Terengganu, Inst Marine Biotechnol, Kuala Nerus, Malaysia. [Wang, Zongling] Minist Nat Resources, Inst Oceanog 1, Key Lab Marine Ecoenvironm Sci & Technol, Qingdao, Peoples R China. [McMinn, Andrew] Univ Tasmania, Inst Marine & Antarctic Studies, Hobart, Tas, Australia. [Wang, Min] Ocean Univ China, Haide Coll, Qingdao, Peoples R China. [Wang, Min] Qingdao Univ, Affiliated Hosp, Qingdao, Peoples R China. RP Liang, YT; McMinn, A; Wang, M (corresponding author), Ocean Univ China, Inst Evolut & Marine Biodivers, Coll Marine Life Sci, Key Lab Polar Oceanog & Global Ocean Change, Qingdao, Peoples R China.; Liang, YT; Wang, M (corresponding author), Univ Malaysia Terengganu Ocean Univ China Joint Ct, Qingdao, Peoples R China.; Wang, ZL (corresponding author), Minist Nat Resources, Inst Oceanog 1, Key Lab Marine Ecoenvironm Sci & Technol, Qingdao, Peoples R China.; McMinn, A (corresponding author), Univ Tasmania, Inst Marine & Antarctic Studies, Hobart, Tas, Australia.; Wang, M (corresponding author), Ocean Univ China, Haide Coll, Qingdao, Peoples R China.; Wang, M (corresponding author), Qingdao Univ, Affiliated Hosp, Qingdao, Peoples R China. 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However, little is known of the microbial ecology of the Andaman Sea and eastern Bay of Bengal (BoB) OMZs. In the present study, a total of 131 samples which from the Andaman Sea and eastern BoB epipelagic waters were analyzed. The microbial community distribution patterns across oxygen gradients, including oxygenic zones (OZs, dissolved oxygen [DO] >= 2 mg/L), oxygen limited zones (OLZs, 0.7 mg/L < DO < 2 mg/L), and OMZs (DO <= 0.7 mg/L), were investigated. Mantel tests and Spearman's correlation analysis revealed that DO was the most important driver of microbial community structures among several environmental factors. Microbial diversity, richness, and evenness were highest in the OLZs and lowest in the OZs. The microbial community compositions of OZ and OMZ waters were significantly different. Random forest analysis revealed 24 bioindicator taxa that differentiated OZ, OLZ, and OMZ water communities. These bioindicator taxa included Burkholderiaceae, HOC36, SAR11 Clade IV, Thioglobaceae, Nitrospinaceae, SAR86, and UBA10353. Further, co-occurrence network analysis revealed that SAR202, AEGEAN-169, UBA10353, SAR406, and Rhodobacteraceae were keystone taxa among the entire interaction network of the microbial communities. Functional prediction further indicated that the relative abundances of microbial populations involved in nitrogen and sulfur cycling were higher in OMZs. Several microbial taxa, including the Thioglobaceae, Nitrospinaceae, SAR202, SAR406, WPS-2, UBA10353, and Woeseiaceae, may be involved in nitrogen and/or sulfur cycling, while also contributing to oxygen consumption in these waters. This study consequently provides new insights into the microbial community structures and potentially important taxa that contribute to oxygen consumption in the Andaman Sea and eastern BoB OMZ. C1 [Guo, Ruoyu; Zhang, Jingjing; Liu, Chenggang; Li, Hongliang; Wang, Pengbin] Minist Nat Resources, Inst Oceanog 2, Key Lab Marine Ecosyst Dynam, Hangzhou, Peoples R China. [Guo, Ruoyu; Ma, Xiao; Zhou, Feng; Wang, Pengbin] Minist Nat Resources, Observat & Res Stn Yangtze River Delta Marine Ecos, Zhoushan, Peoples R China. [Ma, Xiao; Zhou, Feng] Minist Nat Resources, Inst Oceanog 2, State Key Lab Satellite Ocean Environm Dynam, Hangzhou, Peoples R China. [Thu, Chit Aung] Res & Dev Sect, Dept Fisheries, Naypyidaw, Myanmar. [Win, Tun Naing] Minist Transport & Commun, Dept Meteorol & Hydrol, Naypyidaw, Myanmar. [Aung, Nyan Lin] Minist Nat Resources & Environm Conservat, Environm Conservat Dept, Naypyidaw, Myanmar. [Win, Hlaing Swe] Minist Educ, Dept Res Innovat, Natl Analyt Lab, Naypyidaw, Myanmar. [Naing, Sanda] Myanmar Maritime Univ, Port & Harbour Engn Dept, Thanlyin, Myanmar. 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Kuypers, Marcel M. M. Amann, Rudolf TI Potential interactions of particle-associated anammox bacteria with bacterial and archaeal partners in the Namibian upwelling system SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB Recent studies have shown that the anaerobic oxidation of ammonium by anammox bacteria plays an important role in catalyzing the loss of nitrogen from marine oxygen minimum zones (OMZ). However, in situ oxygen concentrations of up to 25 mu M and ammonium concentrations close to or below the detection limit in the layer of anammox activity are hard to reconcile with the current knowledge of the physiology of anammox bacteria. We therefore investigated samples from the Namibian OMZ by comparative 16S rRNA gene analysis and fluorescence in situ hybridization. Our results showed that "Candidatus Scalindua" spp., the typical marine anammox bacteria, colonized microscopic particles that were likely the remains of either macroscopic marine snow particles or resuspended particles. These particles were slightly but significantly (P < 0.01) enriched in Gammaproteobacteria (11.8% +/- 5.0%) compared to the free-water phase (8.1% +/- 1.8%). No preference for the attachment to particles could be observed for members of the Alphaproteobacteria and Bacteroidetes, which were abundant (12 to 17%) in both habitats. The alphaproteobacterial SAR11 clade, the Euryarchaeota, and group I Crenarchaeota, were all significantly depleted in particles compared to their presence in the free-water phase (16.5% +/- 3.5% versus 2.6% +/- 1.7%, 2.7% +/- 1.9% versus < 1%, and 14.9% +/- 4.6% versus 2.2% +/- 1.8%, respectively, all P < 0.001). Sequence analysis of the crenarchaeotal 16S rRNA genes showed a 99% sequence identity to the nitrifying "Nitrosopumilus maritimus." Even though we could not observe conspicuous consortium-like structures of anammox bacteria with particle-enriched bacterioplankton groups, we hypothesize that members of Gammaproteobacteria, Alphaproteobacteria, and Bacteroidetes play a critical role in extending the anammox reaction to nutrient-depleted suboxic water layers in the Namibian upwelling system by creating anoxic, nutrient-enriched microniches. C1 Max Planck Inst Marine Microbiol, D-28359 Bremen, Germany. RP Woebken, D (corresponding author), Max Planck Inst Marine Microbiol, Celsiusstr 1, D-28359 Bremen, Germany. 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TI Characterization of lysogens in bacterioplankton assemblages of the Southern California Borderland SO MICROBIAL ECOLOGY DT Article AB Viruses cause significant mortality of marine microorganisms; however, their role in shaping the composition of microbial assemblages has not been fully elucidated. Because viruses may form lysogenic relationships with their hosts, temperate viruses may influence bacterial assemblage structures through direct lysis of hosts when induced by environmental stimuli or by homoimmunity (i.e., immunity to closely related viruses). We investigated the components of bacterioplankton assemblages that bore prophage using the lysogenic induction agent mitomycin C. Seawater was collected at two locations (the San Pedro Ocean Time Series Station and in the Santa Barbara Channel) in the Southern California Borderland and amended with mitomycin C. 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W. Andersen, Robert A. Gifford, Dian J. Incze, Lewis S. Martin, Jennifer L. Pilskaln, Cynthia H. Rooney-Varga, Juliette N. Sieracki, Michael E. Wilson, William H. Wolff, Nicholas H. TI Planktonic Microbes in the Gulf of Maine Area SO PLOS ONE DT Article AB In the Gulf of Maine area (GoMA), as elsewhere in the ocean, the organisms of greatest numerical abundance are microbes. Viruses in GoMA are largely cyanophages and bacteriophages, including podoviruses which lack tails. There is also evidence of Mimivirus and Chlorovirus in the metagenome. Bacteria in GoMA comprise the dominant SAR11 phylotype cluster, and other abundant phylotypes such as SAR86-like cluster, SAR116-like cluster, Roseobacter, Rhodospirillaceae, Acidomicrobidae, Flavobacteriales, Cytophaga, and unclassified Alphaproteobacteria and Gammaproteobacteria clusters. Bacterial epibionts of the dinoflagellate Alexandrium fundyense include Rhodobacteraceae, Flavobacteriaceae, Cytophaga spp., Sulfitobacter spp., Sphingomonas spp., and unclassified Bacteroidetes. Phototrophic prokaryotes in GoMA include cyanobacteria that contain chlorophyll (mainly Synechococcus), aerobic anoxygenic phototrophs that contain bacteriochlorophyll, and bacteria that contain proteorhodopsin. Eukaryotic microalgae in GoMA include Bacillariophyceae, Dinophyceae, Prymnesiophyceae, Prasinophyceae, Trebouxiophyceae, Cryptophyceae, Dictyochophyceae, Chrysophyceae, Eustigmatophyceae, Pelagophyceae, Synurophyceae, and Xanthophyceae. There are no records of Bolidophyceae, Aurearenophyceae, Raphidophyceae, and Synchromophyceae in GoMA. In total, there are records for 665 names and 229 genera of microalgae. Heterotrophic eukaryotic protists in GoMA include Dinophyceae, Alveolata, Apicomplexa, amoeboid organisms, Labrynthulida, and heterotrophic marine stramenopiles (MAST). Ciliates include Strombidium, Lohmaniella, Tontonia, Strobilidium, Strombidinopsis and the mixotrophs Laboea strobila and Myrionecta rubrum (ex Mesodinium rubra). An inventory of selected microbial groups in each of 14 physiographic regions in GoMA is made by combining information on the depth-dependent variation of cell density and the depth-dependent variation of water volume. Across the entire GoMA, an estimate for the minimum abundance of cell-based microbes is 1.7x10(25) organisms. By one account, this number of microbes implies a richness of 10 5 to 10 6 taxa in the entire water volume of GoMA. Morphological diversity in microplankton is well-described but the true extent of taxonomic diversity, especially in the femtoplankton, picoplankton and nanoplankton - whether autotrophic, heterotrophic, or mixotrophic, is unknown. C1 [Li, William K. W.] Fisheries & Oceans Canada, Bedford Inst Oceanog, Dartmouth, NS B2Y 4A2, Canada. [Andersen, Robert A.; Sieracki, Michael E.; Wilson, William H.] Bigelow Lab Ocean Sci, West Boothbay Harbor, ME USA. [Gifford, Dian J.] Univ Rhode Isl, Grad Sch Oceanog, Narragansett, RI 02882 USA. [Incze, Lewis S.; Wolff, Nicholas H.] Univ So Maine, Aquat Syst Grp, Portland, ME 04103 USA. [Martin, Jennifer L.] Fisheries & Oceans Canada, Biol Stn, St Andrews, NB E0G 2X0, Canada. [Pilskaln, Cynthia H.] Univ Massachusetts, Sch Marine Sci & Technol, N Dartmouth, MA USA. [Rooney-Varga, Juliette N.] Univ Massachusetts, Dept Biol Sci, Lowell, MA USA. RP Li, WKW (corresponding author), Fisheries & Oceans Canada, Bedford Inst Oceanog, POB 1006, Dartmouth, NS B2Y 4A2, Canada. 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Harris, Adeline Ryan, John P. Roman, Brent Marin, Roman, III Jensen, Scott Everlove, Cheri Birch, James Dzenitis, John M. Pargett, Douglas Adachi, Masao Turk, Kendra Zehr, Jonathon P. Scholin, Christopher A. TI Underwater Application of Quantitative PCR on an Ocean Mooring SO PLOS ONE DT Article AB The Environmental Sample Processor (ESP) is a device that allows for the underwater, autonomous application of DNA and protein probe array technologies as a means to remotely identify and quantify, in situ, marine microorganisms and substances they produce. Here, we added functionality to the ESP through the development and incorporation of a module capable of solid-phase nucleic acid extraction and quantitative PCR (qPCR). Samples collected by the instrument were homogenized in a chaotropic buffer compatible with direct detection of ribosomal RNA (rRNA) and nucleic acid purification. From a single sample, both an rRNA community profile and select gene abundances were ascertained. To illustrate this functionality, we focused on bacterioplankton commonly found along the central coast of California and that are known to vary in accordance with different oceanic conditions. DNA probe arrays targeting rRNA revealed the presence of 16S rRNA indicative of marine crenarchaea, SAR11 and marine cyanobacteria; in parallel, qPCR was used to detect 16S rRNA genes from the former two groups and the large subunit RuBisCo gene (rbcL) from Synecchococcus. The PCR-enabled ESP was deployed on a coastal mooring in Monterey Bay for 28 days during the spring-summer upwelling season. The distributions of the targeted bacterioplankon groups were as expected, with the exception of an increase in abundance of marine crenarchaea in anomalous nitrate-rich, low-salinity waters. The unexpected co-occurrence demonstrated the utility of the ESP in detecting novel events relative to previously described distributions of particular bacterioplankton groups. The ESP can easily be configured to detect and enumerate genes and gene products from a wide range of organisms. This study demonstrated for the first time that gene abundances could be assessed autonomously, underwater in near real-time and referenced against prevailing chemical, physical and bulk biological conditions. C1 [Preston, Christina M.; Harris, Adeline; Ryan, John P.; Roman, Brent; Marin, Roman, III; Jensen, Scott; Everlove, Cheri; Birch, James; Pargett, Douglas; Scholin, Christopher A.] Monterey Bay Aquarium Res Inst, Moss Landing, CA 95039 USA. [Dzenitis, John M.] Lawrence Livermore Natl Lab, Livermore, CA USA. [Adachi, Masao] Kochi Univ, Lab Aquat Environm Sci, Kochi 780, Japan. [Turk, Kendra; Zehr, Jonathon P.] Univ Calif Santa Cruz, Dept Ocean Sci & Earth & Marine Sci, Santa Cruz, CA 95064 USA. RP Preston, CM (corresponding author), Monterey Bay Aquarium Res Inst, Moss Landing, CA 95039 USA. 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However, it is unclear how microbial taxa cooccurr and how the inter-taxa networks respond to the strong environmental gradients in the estuaries. Results Here, we constructed co-occurrence networks on prokaryotic microbial communities in the CB, which included seasonal samples from seven spatial stations along the salinity gradients for three consecutive years. Our results showed that spatiotemporal variations of planktonic microbiomes promoted differentiations of the characteristics and stability of prokaryotic microbial networks in the CB estuary. Prokaryotic microbial networks exhibited a clear seasonal pattern where microbes were more closely connected during warm season compared to the associations during cold season. In addition, microbial networks were more stable in the lower Bay (ocean side) than those in the upper Bay (freshwater side). Multivariate regression tree (MRT) analysis and piecewise structural equation modeling (SEM) indicated that temperature, salinity and total suspended substances along with nutrient availability, particulate carbon and Chl a, affected the distribution and co-occurrence of microbial groups, such as Actinobacteria, Bacteroidetes, Cyanobacteria, Planctomycetes, Proteobacteria, and Verrucomicrobia. Interestingly, compared to the abundant groups (such as SAR11, Saprospiraceae and Actinomarinaceae), the rare taxa including OM60 (NOR5) clade (Gammaproteobacteria), Micrococcales (Actinobacteria), and NS11-12 marine group (Bacteroidetes) contributed greatly to the stability of microbial co-occurrence in the Bay. Modularity and cluster structures of microbial networks varied spatiotemporally, which provided valuable insights into the 'small world' (a group of more interconnected species), network stability, and habitat partitioning/preferences. Conclusion Our results shed light on how estuarine gradients alter the spatiotemporal variations of prokaryotic microbial networks in the estuarine ecosystem, as well as their adaptability to environmental disturbances and co-occurrence network complexity and stability. C1 [Wang, Hualong; Wang, Min] Ocean Univ China, Coll Marine Life Sci, Qingdao, Peoples R China. [Wang, Hualong; Wang, Min] Ocean Univ China, Frontiers Sci Ctr Deep Ocean Multi Spheres & Eart, Qingdao, Peoples R China. [Wang, Hualong; Chen, Feng] Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA. [Zhang, Chuanlun] Southern Univ Sci & Technol, Dept Ocean Sci & Engn, Shenzhen, Peoples R China. [Zhang, Chuanlun] Southern Univ Sci & Technol, Shenzhen Key Lab Marine Archaea Geoom, Shenzhen, Peoples R China. [Zhang, Chuanlun] Southern Marine Sci & Engn Guangdong Lab, Guangzhou, Peoples R China. [Kan, Jinjun] Stroud Water Res Ctr, Div Microbiol, Avondale, PA 19311 USA. 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To better understand its influence on microbial community structure and function, we compared the bacterial, archaeal and microeukaryotic communities in the cold water mass area (CWMA) and the southern area (SA) during the summer using amplicon and metagenomic sequencings. The habitat environment in the deep waters of the CWMA was characterized by higher salinity/DO/PO4-P, greater depth/distance to the coast, and lower levels of temperature/chlorophyll a/DIN/SiO3-Si/N:P ratio compared to that of the SA. Pure depth or distance to the coast explained a small portion of the microbial community variance, while environment explained a significant fraction of the variance when partialling the effects of depth and distance to the coast. Oligotrophic taxa (e.g. SAR11 clade Ia, Nitrosopumilus, Chloropicophyceae) dominated the deepwater communities in the CWMA, while the common coastal taxa (e.g. Roseobacter strain HIMB11, Bacillariophyta, Noctilucophyceae) were more dominant in the deep waters of the SA, suggesting the great impact of the oligotrophic condition in the YSCWM on microbial communities. The microbial co-occurrence networks in the CWMA were less complex but contained a higher proportion of mutual exclusion relationship among prokaryotes; the prokaryotic a-diversity in the CWMA was significantly lower than in the SA while the microeukaryotic a-diversity was significantly higher in the CWMA, implying that prokaryotes and microeukaryotes respond to the cold water mass differently and the competition among prokaryotes was intensified under the impact of the YSCWM. 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Gradual microbial community changes induced by the storage and operation of reservoirs were investigated in this study. Microbial 16S rRNA (16S ribosomal RNA) and ITS (Internal Transcribed Spacer) genes were analyzed by high-throughput sequencing to explore the microbial diversity and interaction between microbial species and environmental factors inside and outside reservoirs. The results showed Chao parameters outside the reservoirs were significantly higher than those inside reservoirs, indicating that there are more species outside the reservoir. After the water enters the reservoir, alpha-, beta-, gamma-Proteobacteria decreased in abundance, but the species of Cyanobacteria, Verrucomicrobia and Planctomycete were enriched along with the water flow of the reservoirs. The rising trend of Cyanobacteria in the lower reaches of the reservoir proves the aggravation of eutrophication in the reservoir. Eukaryotic algae of high abundance were also found in the reservoir: Bacillariophyta (diatom), Chlorophyta (green algae), and Cryptomonadaceae. Less amount of these eukaryotic algae observed outside the reservoir further confirmed the eutrophication tendency inside the reservoir. Therefore, as the prolongation of residence time in the reservoir, the degree of eutrophication was enhanced accompanied by the directional transition of community structures. It was found that the decrease abundance of methylophilaceae (aerobic denitrification), pelagibacter and xylophilus (aerobic oxidation) occurred simultaneously with the increasing concentrations of CODMn, TOC and NO3-N after entering the reservoir. Actinobacteria was found to be the most dominant population. Some Fungi enriched in the reservoir such as Chytridiomycota, Sclerotiniaceae, and Rhizophydium are related to algae proliferations, which may increase the risk of forming water odor. The degree of decay in the reservoir is considered to be increasing due to rapid growth of both Bacteroidia and Chytridiomycota, which are supposed to be indicator bacteria for decaying. The detention area in the reservoir is likely to be an important driving factor. The research confirmed the influence factors of eutrophication and the ecological risk of the reservoir. The discovery of some functional microorganisms in the reservoir help understand the relationship between the ecological effect and the safety of drinking water source. C1 [Guo, Jun; Zheng, Yuanyuan; Teng, Jinhao; Wang, Xumin; Song, Jiaxiu] Shanghai Normal Univ, Sch Environm & Geog Sci, Dept Environm Engn, Shanghai 200234, Peoples R China. [Guo, Jun; Zheng, Yuanyuan; Teng, Jinhao; Wang, Xumin; Song, Jiaxiu] Wetland Ecosyst Observat & Res Field Stn, Shanghai 201799, Peoples R China. RP Song, JX (corresponding author), Shanghai Normal Univ, Sch Environm & Geog Sci, Dept Environm Engn, Shanghai 200234, Peoples R China. 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Gross and net growth rates of different bacterioplankton groups targeted by group-specific CARD-FISH probes and infrared microscopy (for aerobic anoxygenic phototrophs, AAP), were calculated from changes in cell abundances. Maximal group-specific growth rates were achieved when both predation pressure and nutrient limitation were experimentally minimized, while only a minimal effect of viral pressure on growth rates was observed; nevertheless, the response to predation removal was more remarkable in winter, when the bacterial community was not subjected to nutrient limitation. Although all groups showed increases in their growth rates when resource competition as well as grazers and viral pressure were reduced, Alteromonadaceae consistently presented the highest rates in all seasons. The response to light availability was generally weaker than that to the other factors, but it was variable between seasons. In summer and spring, the growth rates of AAP were stimulated by light whereas the growth of the SAR11 clade (likely containing proteorhodopsin) was enhanced by light in all seasons. Overall, our results set thresholds on bacterioplankton group-specific growth and mortality rates and contribute to estimate the seasonally changing contribution of various bacterioplankton groups to the function of microbial communities. Our results also indicate that the least abundant groups display the highest growth rates, contributing to the recycling of organic matter to a much greater extent than what their abundances alone would predict. C1 [Sanchez, Olga; Mabrito, Isabel; Gazulla, Carlota R.] Univ Autonoma Barcelona, Dept Genet & Microbiol, Bellaterra 08193, Catalunya, Spain. [Ferrera, Isabel] Inst Espanol Oceanog, Ctr Oceanog Malaga, Malaga 29640, Spain. [Ferrera, Isabel; Gazulla, Carlota R.; Sebastian, Marta; Auladell, Adria; Marin-Vindas, Carolina; Cardelus, Clara; Sanz-Saez, Isabel; Pernice, Massimo C.; Marrase, Celia; Sala, M. Montserrat; Gasol, Josep M.] ICM CSIC, Dept Biol Marina & Oceanog, Inst Ciencies Mar, Barcelona 08003, Catalunya, Spain. [Sebastian, Marta] Univ Las Palmas de Gran Canaria ULPGC, Inst Oceanog & Cambio Global IOCAG, Telde 35214, Spain. [Marin-Vindas, Carolina] Univ Nacional, Escuela Ciencias Biol, Heredia 40101, Costa Rica. RP Sánchez, O (corresponding author), Univ Autonoma Barcelona, Dept Genet & Microbiol, Bellaterra 08193, Catalunya, Spain.; Ferrera, I (corresponding author), Inst Espanol Oceanog, Ctr Oceanog Malaga, Malaga 29640, Spain.; Ferrera, I (corresponding author), ICM CSIC, Dept Biol Marina & Oceanog, Inst Ciencies Mar, Barcelona 08003, Catalunya, Spain. 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Zhao, Liang Stemple, Brooke DeLong, Kimberly Medeiros, Patricia M. Seim, Harvey Marchetti, Adrian TI Microbial Niche Diversification in the Galapagos Archipelago and Its Response to El Nino SO FRONTIERS IN MICROBIOLOGY DT Article AB The Galapagos Archipelago is located at the intersection of several major oceanographic features that produce diverse environmental conditions around the islands, and thus has the potential to serve as a natural laboratory for discerning the underlying environmental factors that structure marine microbial communities. Here we used quantitative metagenomics to characterize microbial communities in relation to archipelago marine habitats, and how those populations shift due to substantial environmental changes brought on by El Nino. Environmental conditions such as temperature, salinity, inorganic dissolved nutrients, and dissolved organic carbon (DOC) concentrations varied throughout the archipelago, revealing a diversity of potential microbial niches arising from upwelling, oligotrophic to eutrophic gradients, physical isolation, and potential island mass effects. The volumetric abundances of microbial community members shifted with these environmental changes and revealed several taxonomic indicators of different water masses. This included a transition from a Synechococcus dominated system in the west to an even mix of Synechococcus and Prochlorococcus in the east, mirroring the archipelago's mesotrophic to oligotrophic and productivity gradients. Several flavobacteria groups displayed characteristic habitat distributions, including enrichment of Polaribacter and Tenacibaculum clades in the relatively nutrient rich western waters, Leeuwenhoekiella spp. that were enriched in the more nutrient-deplete central and eastern sites, and the streamlined MS024-2A group found to be abundant across all sites. During the 2015/16 El Nino event, both environmental conditions and microbial community composition were substantially altered, primarily on the western side of the archipelago due to the reduction of upwelling from the Equatorial Undercurrent. When the upwelling resumed, concentrations of inorganic nutrients and DOC at the western surface sites were more typical of mesopelagic depths. Correspondingly, Synechococcus abundances decreased by an order of magnitude, while groups associated with deeper water masses were enriched, including streamlined roseobacters HTCC2255 and HIMB11, Thioglobacaceae, methylotrophs (Methylophilaceae), archaea (Nitrosopumilaceae), and distinct subpopulations of Pelagibaceriales (SAR11 clade). These results provide a quantitative framework to connect community-wide microbial volumetric abundances to their environmental drivers, and thus incorporation into biogeochemical and ecological models. C1 [Gifford, Scott M.; Zhao, Liang; Seim, Harvey; Marchetti, Adrian] Univ N Carolina, Dept Marine Sci, Chapel Hill, NC 27515 USA. [Stemple, Brooke] Univ Notre Dame, Dept Civil & Environm Engn & Earth Sci, Notre Dame, IN 46556 USA. [DeLong, Kimberly] Univ Calif Santa Cruz, Dept Ocean Sci, Santa Cruz, CA 95064 USA. [Medeiros, Patricia M.] Univ Georgia, Dept Marine Sci, Athens, GA 30602 USA. 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We additionally demonstrated that modifications to the 515f/806r (variable region 4) 16S primer pair, which improves detection of Thaumarchaeota and clade SAR11 in marine samples, do not degrade performance on taxa already amplified effectively by the original primer set. Alterations to the fungal ITS primers did result in differential but overall improved performance compared to the original primers. In both cases, the improved primers should be widely adopted for amplicon studies. IMPORTANCE We continue to uncover a wealth of information connecting microbes in important ways to human and environmental ecology. As our scientific knowledge and technical abilities improve, the tools used for microbiome surveys can be modified to improve the accuracy of our techniques, ensuring that we can continue to identify groundbreaking connections between microbes and the ecosystems they populate, from ice caps to the human body. It is important to confirm that modifications to these tools do not cause new, detrimental biases that would inhibit the field rather than continue to move it forward. We therefore demonstrated that two recently modified primer pairs that target taxonomically discriminatory regions of bacterial and fungal genomic DNA do not introduce new biases when used on a variety of sample types, from soil to human skin. This confirms the utility of these primers for maintaining currently recommended microbiome research techniques as the state of the art. C1 [Walters, William] Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY USA. [Hyde, Embriette R.; Ackermann, Gail; Humphrey, Greg; Knight, Rob] Univ Calif San Diego, Dept Pediat, La Jolla, CA 92093 USA. [Berg-Lyons, Donna] Univ Colorado, BioFrontiers Inst, Boulder, CO 80309 USA. [Parada, Alma; Fuhrman, Jed A.] Univ Southern Calif, Dept Biol Sci, Los Angeles, CA 90089 USA. [Gilbert, Jack A.] Argonne Natl Lab, Biosci Div BIO, 9700 S Cass Ave, Argonne, IL 60439 USA. [Gilbert, Jack A.] Univ Chicago, Dept Ecol & Evolut, 940 E 57th St, Chicago, IL 60637 USA. [Gilbert, Jack A.] Univ Chicago, Dept Surg, 5841 S Maryland Ave, Chicago, IL 60637 USA. [Gilbert, Jack A.] Univ Chicago, Inst Genom & Syst Biol, Chicago, IL 60637 USA. [Gilbert, Jack A.] Marine Biol Lab, Woods Hole, MA 02543 USA. [Gilbert, Jack A.] Field Museum Nat Hist, Chicago, IL 60605 USA. [Jansson, Janet K.] Pacific Northwest Natl Lab, Earth & Biol Sci Directorate, Richland, WA 99352 USA. [Caporaso, J. Gregory] No Arizona Univ, Dept Biol Sci, Box 5640, Flagstaff, AZ 86011 USA. [Apprill, Amy] Woods Hole Oceanog Inst, Dept Marine Chem & Geochem, Woods Hole, MA 02543 USA. [Knight, Rob] Univ Calif San Diego, Dept Comp Sci, La Jolla, CA 92093 USA. [Knight, Rob] Univ Calif San Diego, Dept Engn, La Jolla, CA 92093 USA. RP Knight, R (corresponding author), Univ Calif San Diego, Dept Pediat, La Jolla, CA 92093 USA.; Knight, R (corresponding author), Univ Calif San Diego, Dept Comp Sci, La Jolla, CA 92093 USA.; Knight, R (corresponding author), Univ Calif San Diego, Dept Engn, La Jolla, CA 92093 USA. 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This article reviews marine fungal diversity in the light of current knowledge, citing examples of how progress in understanding marine prokaryotes has often contributed to this new approach. Both 'true fungi' (termed mycenaean fungi in this review) and straminipilan fungi are considered. Molecular phylogenetic studies of prokaryotes has resulted in their redefinition as belonging to the Kingdoms Bacteria and Archaea. Likewise, major refinements have taken place in the phylogenetic classification of eukaryotes. In the case of fungi, it has now been realized that they are polyphyletic, belonging to the Kingdom Mycenae (Fungi), as well as the Kingdom Strairtinipila or Chromista. Although the total number of fungi on earth is estimated to be about 1.5 million, only a meagre number of obligate marine fungi, about 450 mycenaean and 50 straminipilan fungi have been described so far. It is likely that most of the true marine fungi have not yet been discovered. These are likely to have evolved between 1,500 million years ago (Ma) when fungi probably evolved in the sea and 900 Ma when they conquered land together with green plants. It now appears that most of the true marine fungi have not been cultured so far, similar to the 'great plate count anomaly' of bacteria. Thraustochytrids, which are abundant in the water column, but not easily culturable from that source is an example. Intelligent and novel culture methods might bring forth unusual and new marine fungi, as happened in the case of Pelagibacter ubique belonging to the SAR 11 group of bacteria. Molecular techniques might bring to light novel marine fungi, as is happening with bacteria. Such fungi may defy our conventional wisdom regarding these organisms in terms of morphology. Thus, several recent studies using 18S rRNA gene community profiles have discovered picoplanktonic marine fungi in the water column. Studies such as those on molecular diversity of eukaryotes in permanently anoxic habitats have also indicated that fungi may be abundant in exotic habitats and possess unusual physiology. A search for fungi in biodiversity-rich habitats, such as the coral reefs and the deep-sea, using a combination of molecular and novel culture methods is likely to reveal a fascinating diversity of marine fungi. C1 Myko Tech Pvt Ltd, Panaji 403004, Goa, India. RP Raghukumar, S (corresponding author), Myko Tech Pvt Ltd, 313 Vainguinnim Valley, Panaji 403004, Goa, India. 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Due to the unique hydrological, geological and geomorphological features, the basin is a biodiversity hot-spot of multicellular organisms, and likewise can harbor peculiar assemblages of microbes if compared to other oceans. However, we still know little about the diversity of Mediterranean microbes, due to past methodological constraints only recently conquerable with next generation sequencing of DNA and metagenomics. This review aims at summarizing the knowledge on microbial diversity in the basin, by focusing on prokaryotes (bacteria and archaea) in pelagic and benthic habitats. Richness of bacterioplankton and archaeoplankton assemblages is high in surface and deep waters, and shows consistent horizontal and vertical patterns. 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S. van de Poll, Willem H. Eveleth, Rachel Rich, Jeremy J. Ducklow, Hugh W. Rozema, Patrick D. Luria, Catherine M. Bolhuis, Henk Meredith, Michael P. Amaral-Zettler, Linda A. Engelmann, Julia C. TI Spatial and temporal variation of Antarctic microbial interactions: a study around the west Antarctic Peninsula SO ENVIRONMENTAL MICROBIOME DT Article AB Background The west Antarctic Peninsula (WAP) is a region of rapid environmental changes, with regional differences in climate warming along the north-south axis of the peninsula. Along the WAP, Palmer corresponds to a warmer region with lesser sea ice extent in the north compared to Rothera similar to 400 km to the south. Comprehensive and comparative, year-round assessments of the WAP microbial community dynamics in coastal surface waters at these two locations are imperative to understand the effects of regional climate warming variations on microbial community dynamics, but this is still lacking. Results We report on the seasonal diversity, taxonomic overview, as well as predicted inter-and intra-domain causal effects (interactions) of the bacterial and microbial eukaryotic communities close to the Palmer station and at the Rothera time-series site between July 2013 and April 2014. Our 16S- and 18S-rRNA gene amplicon sequencing data showed that across all seasons, both bacteria and microbial eukaryotic communities were considerably different between the two sites which could be attributed to seawater temperature, and sea ice coverage in combination with sea ice type differences. Overall, in terms of biotic drivers, causal-effect modelling suggests that bacteria were stronger drivers of ecosystem dynamics at Palmer, while microbial eukaryotes played a stronger role at Rothera. The parasitic taxa Syndiniales persevered at both sites across the seasons, with Palmer and Rothera harbouring different key groups. Up to 62.3% of the negative causal effects were driven by Syndiniales at Rothera compared to only 13.5% at Palmer, suggesting that parasitism drives community dynamics at Rothera more strongly than at Palmer. Conversely, SAR11 Clade II, which was less abundant but persistent year-round at both sites, was the dominant driver at Palmer, evidenced by many (28.2% and 37.4% of positive and negative effects respectively) strong causal effects. Article note: Kindly check first page article notes are correct. Conclusions Our research has shed light on the dynamics of microbial community composition and correlative interactions at two sampling locations that represent different climate regimes along the WAP. C1 [Sow, Swan L. S.; Bolhuis, Henk; Amaral-Zettler, Linda A.; Engelmann, Julia C.] NIOZ Royal Netherlands Inst Sea Res, Dept Marine Microbiol & Biogeochem, POB 59, NL-1790 AB Den Burg, Netherlands. [van de Poll, Willem H.; Rozema, Patrick D.] Univ Groningen, CIO Oceans Energy & Sustainabil Res Inst Groningen, Fac Sci & Engn, Groningen, Netherlands. [Eveleth, Rachel] Oberlin Coll, Dept Geosci, Oberlin, OH USA. [Rich, Jeremy J.] Univ Maine, Darling Marine Ctr, Sch Marine Sci, Walpole, ME USA. [Ducklow, Hugh W.] Columbia Univ, Dept Earth & Environm Sci, New York, NY USA. [Ducklow, Hugh W.] Columbia Univ, Lamont Doherty Earth Observ, Palisades, NY USA. [Luria, Catherine M.] Harvard Med Sch, Lab Syst Pharmacol, Boston, MA USA. [Meredith, Michael P.] British Antarctic Survey, Cambridge, England. [Amaral-Zettler, Linda A.] Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, Dept Freshwater & Marine Ecol, Amsterdam, Netherlands. [Amaral-Zettler, Linda A.] Josephine Bay Paul Ctr, Marine Biol Lab, Woods Hole, MA 02543 USA. [Sow, Swan L. S.] Nantes Univ, Ecole Cent Nantes, CNRS, UMR 6004,LS2N, F-44000 Nantes, France. RP Sow, SLS; Amaral-Zettler, LA; Engelmann, JC (corresponding author), NIOZ Royal Netherlands Inst Sea Res, Dept Marine Microbiol & Biogeochem, POB 59, NL-1790 AB Den Burg, Netherlands.; Amaral-Zettler, LA (corresponding author), Univ Amsterdam, Inst Biodivers & Ecosyst Dynam, Dept Freshwater & Marine Ecol, Amsterdam, Netherlands.; Amaral-Zettler, LA (corresponding author), Josephine Bay Paul Ctr, Marine Biol Lab, Woods Hole, MA 02543 USA.; Sow, SLS (corresponding author), Nantes Univ, Ecole Cent Nantes, CNRS, UMR 6004,LS2N, F-44000 Nantes, France. 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Xia, Li C. TI Explore mediated co-varying dynamics in microbial community using integrated local similarity and liquid association analysis SO BMC GENOMICS DT Article; Proceedings Paper CT 17th Asia Pacific Bioinformatics Conference (APBC) - Genomics CY JAN 16, 2019 CL Wuhan, PEOPLES R CHINA AB BackgroundDiscovering the key microbial species and environmental factors of microbial community and characterizing their relationships with other members are critical to ecosystem studies. The microbial co-occurrence patterns across a variety of environmental settings have been extensively characterized. However, previous studies were limited by their restriction toward pairwise relationships, while there was ample evidence of third-party mediated co-occurrence in microbial communities.MethodsWe implemented and applied the triplet-based liquid association analysis in combination with the local similarity analysis procedure to microbial ecology data. We developed an intuitive scheme to visualize those complex triplet associations along with pairwise correlations. Using a time series from the marine microbial ecosystem as example, we identified pairs of operational taxonomic units (OTUs) where the strength of their associations appeared to relate to the values of a third mediator variable. These mediator variables appear to modulate the associations between pairs of bacteria.ResultsUsing this analysis, we were able to assess the OTUs' ability to regulate its functional partners in the community, typically not manifested in the pairwise correlation patterns. For example, we identified Flavobacteria as a multifaceted player in the marine microbial ecosystem, and its clades were involved in mediating other OTU pairs. By contrast, SAR11 clades were not active mediators of the community, despite being abundant and highly correlated with other OTUs. Our results suggested that Flavobacteria are more likely to respond to situations where particles and unusual sources of dissolved organic material are prevalent, such as after a plankton bloom. On the other hand, SAR11s are oligotrophic chemoheterotrophs with inflexible metabolisms, and their relationships with other organisms may be less governed by environmental or biological factors.ConclusionsBy integrating liquid association with local similarity analysis to explore the mediated co-varying dynamics, we presented a novel perspective and a useful toolkit to analyze and interpret time series data from microbial community. Our augmented association network analysis is thus more representative of the true underlying dynamic structure of the microbial community. The analytic software in this study was implemented as new functionalities of the ELSA (Extended local similarity analysis) tool, which is available for free download (http://bitbucket.org/charade/elsa). C1 [Ai, Dongmei; Li, Xiaoxin; Pan, Hongfei] Univ Sci & Technol Beijing, Sch Math & Phys, Xueyuan Rd, Beijing 100001, Peoples R China. [Chen, Jiamin; Xia, Li C.] Stanford Univ, Sch Med, Dept Med, 269 Campus Dr, Stanford, CA 94305 USA. [Cram, Jacob A.] Univ Maryland, Ctr Environm Sci, Cambridge, MA 21613 USA. RP Xia, LC (corresponding author), Stanford Univ, Sch Med, Dept Med, 269 Campus Dr, Stanford, CA 94305 USA.; Cram, JA (corresponding author), Univ Maryland, Ctr Environm Sci, Cambridge, MA 21613 USA. 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The majority of microorganisms were confined to discreet horizontal layers of no more than 30 m (stenobathic). Only a few such as members of the SAR11 clade appeared at all depths (eurybathic). During the winter mixing period, only some groups of bloomers such as Pseudomonas were favored. Although most microbes appeared in both seasons, some groups like the SAR116 clade and some Bacteroidetes and Verrucomicrobia seemed to disappear during the mixing period. Furthermore, we found that some microbes previously considered seasonal (e.g., Archaea or Actinobacteria) were living in deeper layers within the photic zone during the stratification period. A strong depth-related specialization was detected, not only at the taxonomic level but also at the functional level, even within the different clades, for the manipulation and uptake of specific polysaccharides. 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McGovern, Maeve Gradinger, Rolf Praebel, Kim Poste, Amanda E. TI Terrestrial Inputs Shape Coastal Bacterial and Archaeal Communities in a High Arctic Fjord (Isfjorden, Svalbard) SO FRONTIERS IN MICROBIOLOGY DT Article AB The Arctic is experiencing dramatic changes including increases in precipitation, glacial melt, and permafrost thaw, resulting in increasing freshwater runoff to coastal waters. During the melt season, terrestrial runoff delivers carbon- and nutrient-rich freshwater to Arctic coastal waters, with unknown consequences for the microbial communities that play a key role in determining the cycling and fate of terrestrial matter at the land-ocean interface. To determine the impacts of runoff on coastal microbial (bacteria and archaea) communities, we investigated changes in pelagic microbial community structure between the early (June) and late (August) melt season in 2018 in the Isfjorden system (Svalbard). Amplicon sequences of the 16S rRNA gene were generated from water column, river and sediment samples collected in Isfjorden along fjord transects from shallow river estuaries and glacier fronts to the outer fjord. Community shifts were investigated in relation to environmental gradients, and compared to river and marine sediment microbial communities. We identified strong temporal and spatial reorganizations in the structure and composition of microbial communities during the summer months in relation to environmental conditions. Microbial diversity patterns highlighted a reorganization from rich communities in June toward more even and less rich communities in August. In June, waters enriched in dissolved organic carbon (DOC) provided a niche for copiotrophic taxa including Sulfitobacter and Octadecabacter. 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Autotrophic and heterotrophic picoplankton abundance and community composition with a focus on picoeukaryotes (PEs) were investigated in the winter of 2016 at three stations along a coast-to-offshore transect in the southern Adriatic Sea. Abundances were estimated by flow cytometry, while community composition by Illumina High Throughput Sequencing (HTS) of 16S and 18S rRNA genes. The photosynthetic picoplankton diversity was also investigated by High-Performance Liquid Chromatography (HPLC) of liposoluble pigments. Heterotrophic bacteria and cyanobacteria (Prochlorococcus and Synechococcus) accounted for up to 7 x 10(5); 2.3 x 10(4) and 2.5 x 10(4) cells mL(-1), respectively, while photosynthetic picoeukaryotes peaked with 3 x 10(3) cells mL(-1). Prokaryotes, as revealed by HTS were dominated by Aiphaproteobacteria (mainly SAR11, 44.91% of total 16S sequence reads), followed by Gammaproteobacteria (Oceanospirillales and Pseudomonadales, 14.96%), Bacteroidetes (mainly Flavobacteriales, 13%), Cyanobacteria (Prochlorococcus and Synechococcus, 9.52%), Marinimicrobia (SAR406, 7.97%), Deltaproteobacteria (SAR324, 3.83%), Actinobacteria (2.24%) and Chloroflezi (SAR202, 1.90%). Photosynthetic pigment concentrations were very low (12.12 mu gL(-1) at the most) and taxonomic pigments could be attributed to Prochlorococcus, Synechococcus, Prymnesiophyceae, Bacillariophyceae, Chrysophyceae, and Prasinophyceae. HTS data revealed that PEs were dominated by heterotrophs, such as Syndiniophyceae, parasitic dinoflagellates (79.67% of total 18S sequence reads), Dinophyceae (8.7%) and the radiolarians Collodaria belonging to Sphaerozoidae (22.1%) and Spumellaria (5.0%). 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and Yellow Seas SO JOURNAL OF GEOPHYSICAL RESEARCH-OCEANS DT Article AB Dimethyl sulfide (DMS) and its precursor dimethylsulfoniopropionate (DMSP) are ubiquitous sulfur compounds in the ocean. DMS is emitted into the atmosphere and has potential climatic effects. The distributions of DMS and DMSP are affected by various biological factors (i.e., bacterial catabolism and phytoplankton and zooplankton community composition). The horizontal and vertical distributions of DMSP, DMSP lyase activity (DLA), DMS, and the abundances of bacteria, DMSP-consuming bacteria, dimethyl sulfoxide-consuming bacteria, and picophytoplankton were investigated in the Bohai Sea (BS) and Yellow Sea (YS) during autumn 2020. DLA was significantly correlated with chlorophyll a, DMS, and dissolved DMSP concentrations. Our data show that bacteria Clade_I SAR11 was a significant contributor to DLA. A dilution experiment indicated that the highest microzooplankton grazing rate coincided with the highest DMS concentration and DMS production rate. A proportion of 16%-62% of the DMSPt was converted to DMS in the dilution experiment. Copepods dominated the mesozooplankton community. Calanus sinicus was the predominant copepod in the BS and YS. C. sinicus grazing stimulated DMS production. DMS concentration increased 299% after C. sinicus grazing on physically broken algal cells for 48 hr. These results will help with a better understanding of the control of DMS and DMSP concentrations by zooplankton and the DMS release mechanisms that occur through zooplankton grazing.Plain Language Summary Dimethylsulfoniopropionate (DMSP) and dimethyl sulfide (DMS) are transferred and transformed in the food web and are affected by zooplankton, phytoplankton, and bacteria. This study investigated spatial distributions of DMS, DMSP, DMSP lyase activity (DLA), the abundances of bacteria, DMSP-consuming bacteria, and dimethyl sulfoxide-consuming bacteria in the surface seawater of the Bohai Sea and Yellow Sea. As results, the distributions of DMS and dissolved DMSP were significantly correlated with DLA. A deck incubation experiment indicated that grazing by microzooplankton promoted the conversion of DMSP to DMS and that mesozooplankton copepod grazing stimulated DMS production. These results provide a more comprehensive way to understand the possible DMS release mechanisms via zooplankton grazing. C1 [Yu, Juan; Wang, Su; Lai, Jing-Guang; Zhang, Hao-Quan; Yang, Gui-Peng; Chen, Rong] Ocean Univ China, Coll Chem & Chem Engn, Frontiers Sci Ctr Deep Ocean Multispheres & Earth, Key Lab Marine Chem Theory & Technol, Qingdao, Peoples R China. [Yu, Juan; Yang, Gui-Peng] Qingdao Natl Lab Marine Sci & Technol, Lab Marine Ecol & Environm Sci, Qingdao, Peoples R China. [Tian, Ji-Yuan] Qingdao Agr Univ, Coll Food Sci & Engn, Qingdao, Peoples R China. 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In this study, we present the results of a 4-year study conducted in a well-characterized coral reef ecosystem (Paopao Bay, Moorea, French Polynesia) where changes in bacterioplankton abundance and dissolved organic carbon (DOC) concentrations were quantified and bacterial community structure variation was examined along spatial gradients of the reef: ocean interface. Our results illustrate that the reef is consistently depleted in concentrations of both DOC and bacterioplankton relative to offshore waters (averaging 79 mu mol l(-1) DOC and 5.5 x 10(8) cells l(-1) offshore and 68 lmol l(-1) DOC and 3.1 x 10(8) cells l(-1) over the reef, respectively) across a 4-year time period. In addition, using a suite of culture-independent measures of bacterial community structure, we found consistent differentiation of reef bacterioplankton communities from those offshore or in a nearby embayment across all taxonomic levels. Reef habitats were enriched in Gamma-, Delta-, and Betaproteobacteria, Bacteriodetes, Actinobacteria and Firmicutes. Specific bacterial phylotypes, including members of the SAR11, SAR116, Flavobacteria, and Synechococcus clades, exhibited clear gradients in relative abundance among nearshore habitats. Our observations indicate that this reef system removes oceanic DOC and exerts selective pressures on bacterioplankton community structure on timescales approximating reef water residence times, observations which are notable both because fringing reefs do not exhibit long residence times (unlike those characteristic of atoll lagoons) and because oceanic DOC is generally recalcitrant to degradation by ambient microbial assemblages. Our findings thus have interesting implications for the role of oceanic DOM and bacterioplankton in the ecology and metabolism of reef ecosystems. The ISME Journal (2011) 5, 1374-1387; doi:10.1038/ismej.2011.12; published online 10 March 2011 C1 [Nelson, Craig E.; Alldredge, Alice L.; Carlson, Craig A.] Univ Calif Santa Barbara, Inst Marine Sci, Santa Barbara, CA 93106 USA. [Alldredge, Alice L.; Carlson, Craig A.] Univ Calif Santa Barbara, Dept Ecol Evolut & Marine Biol, Santa Barbara, CA 93106 USA. [McCliment, Elizabeth A.; Amaral-Zettler, Linda A.] Marine Biol Lab, Josephine Bay Paul Ctr Comparat Mol Biol & Evolut, Woods Hole, MA 02543 USA. [Amaral-Zettler, Linda A.] Brown Univ, Dept Geol Sci, Providence, RI 02912 USA. RP Nelson, CE (corresponding author), Univ Calif Santa Barbara, Inst Marine Sci, Santa Barbara, CA 93106 USA. 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Near the vent orifice, representing the initial stage of plume formation, microbial populations were characterized by abundant and diverse putative vent-associated communities including (hyper)thermophiles such as Aquificaceae and Hydrogenothermaceae within the phylum Aquificae, and some epsilonproteobacterial chemolithoautotrophs such as Sulfurovum, Sulfurimonas, and Caminibacter. By contrast, in the rising buoyant plumes and adjacent seawaters, most vent-associated microbial taxa were still present but made only minor contributions to community composition. Some microbial taxa that are common in seawater columns such as alphaproteobacterial Sphingomonadaceae and SAR11 clade, deltaproteobacterial SAR324 clade, and gammaproteobacterial Pseudomonas, together with Sulfurimonas and SUP05 clade, became predominant. 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10.3389/fmars.2020.581381 UT WOS:000591538300001 DA 2025-07-30 ER PT J AU Coutinho, FH Rosselli, R Rodríguez-Valera, F AF Hernandes Coutinho, Felipe Rosselli, Riccardo Rodriguez-Valera, Francisco TI Trends of Microdiversity Reveal Depth-Dependent Evolutionary Strategies of Viruses in the Mediterranean SO MSYSTEMS DT Article AB The evolutionary interactions between viruses and their prokaryotic hosts remain a little-known aspect of microbial evolution. Most studies on this topic were carried out in pure cultures that challenge one virus with one bacterial clone at a time, which is very removed from real-life situations. Few studies have addressed trends of microdiversity in marine viral communities throughout depth gradients. We analyzed metagenomes from both the cellular and viral fractions of Mediterranean seawater samples spanning the epipelagic to the bathypelagic zones at depths of 15, 45, 60, and 2,000 m during the summer stratification of the water column. We evaluated microdiversity patterns by measuring the accumulation of synonymous and nonsynonymous mutations in viral genes. Our results demonstrated clear depth-dependent trends in the frequency of polymorphic sites and nonsynonymous mutations among genes encoding metabolic, structural, and replication proteins. These differences were linked to changes in energy availability, host and viral densities, and the proportions of actively replicating viruses. We propose the hypothesis that in the energy-rich, high-host-density, euphotic depths, selection acts to favor diversity of the host recognition machinery to increase host range, while in energy-depleted aphotic waters, selection acts on viral replication fitness, enhancing diversity in auxiliary metabolic genes. IMPORTANCE Viruses are extremely abundant and diverse biological entities that contribute to the functioning of marine ecosystems. Despite their recognized importance, few studies have addressed trends of mutation accumulation in marine viral communities across depth gradients. By investigating these trends, we show that mutation frequencies differ among viral genes according to their molecular functions, with the highest microdiversity occurring among proteins related to host metabolism, followed by structural proteins and, lastly, genome replication proteins. This is in agreement with evolutionary theory that postulates that housekeeping genes are under strong purifying selection. We also observed a positive association between depth and microdiversity. One exception to this trend was the host recognition proteins from the deep chlorophyll maximum, which displayed strikingly high microdiversity, which we hypothesize to be associated with intraspecies competition for hosts. Finally, our data allowed us to propose a theoretical model for viral micro-diversity across the depth gradient. These discoveries are of special relevance because many of the viral genomic sequences discovered here were predicted to infect some of the most abundant bacteria in marine ecosystems, such as "Candidatus Pelagibacter," Puniceispirillum, and Prochlorococcus. C1 [Hernandes Coutinho, Felipe; Rosselli, Riccardo; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Dept Prod Vegetal & Microbiol, Evolutionary Genom Grp, Campus San Juan, Alicante, Spain. [Rodriguez-Valera, Francisco] Moscow Inst Phys & Technol, Dolgoprudnyi, Russia. RP Coutinho, FH (corresponding author), Univ Miguel Hernandez, Dept Prod Vegetal & Microbiol, Evolutionary Genom Grp, Campus San Juan, Alicante, Spain. 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Bacterial denitrifiers (3.7 +/- 6.9 bacterial nirK gene mL(-1)) and anaerobic ammonia oxidizers (78 +/- 322 anammox 16S rRNA genes L-1) were low in abundance under the oxygen conditions in the Gulf of Alaska throughout the water column. The widespread distribution of bacterial denitrifiers and anaerobic ammonia oxidizers in low abundances reveals a reservoir of genetic and metabolic potential ready to colonize the environment under the predicted increase of OMZs in the ocean. Taken together, our results reinforce the niche partitioning of archaeal ammonia oxidizers based on their distinct metabolic characteristics resulting in the dominance of LAC-AOA in a high latitude deep OMZ. Considering the different ecological roles and functions of the two archaeal ecotypes, the expansion of the zones dominated by the LAC-ecotype might have implications for the nitrogen cycle in the future ocean. 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This study investigates the role of the organic and inorganic nitrogen and phosphorus compounds released by copepods on biogeochemical processes and on the microbial community composition during the OUTPACE cruise (18 February-3 April 2015) at three longduration stations (LD). Two LD stations were located in the Melanesian Archipelago region (MA; LD A and LD B) and one in the South Pacific Gyre (SG; LD C), which represent oligotrophic and ultra-oligotrophic regions respectively. At each station, on-board microcosm experiments were performed with locally sampled organisms, comprising a mix of epipelagic copepods fed with their natural food and then incubated along with wild microbial assemblages. In the presence of copepods, ammonium and dissolved organic nitrogen showed a significant increase compared to a control in two situations: in ammonium concentration (rate: 0.29 mu mol L-1 h(-1) after 4 h of incubation) in LD C and in dissolved organic nitrogen concentration (rate: 2.13 mu mol L-1 h(-1) after 0.5 h of incubation) in LD A. In addition, during the three experiments, an enhanced remineralization (ammonification and nitrification) was observed when adding copepods compared to the controls. A shift in the composition of the active bacterial community was observed for the experiments in LD A and LD B, which were mainly characterized by an increase in Alteromonadales and SAR11, respectively, and linked with changes in nutrient concentrations. In the experiment performed in LD C, both groups increased but at different periods of incubation. Alteromonadales increased between 1 and 2 h after the beginning of the experiment, and SAR 11 at the end of incubation. Our results in near in situ conditions show that copepods can be a source of organic and inorganic compounds for bacterial communities, which respond to excretion pulses at different timescales, depending on the initial environmental conditions and on their community composition. These processes can significantly contribute to nutrient recycling and regenerated production in the photic zone of ultra-oligotrophic and oligotrophic oceanic regions. C1 [Valdes, Valentina] Univ Concepcion, Fac Ciencias Nat & Oceanog, Dept Oceanog, Programa Doctorado Oceanog, Concepcion, Chile. [Valdes, Valentina; Fernandez, Camila] UPMC Univ Paris 06, Sorbonne Univ, Lab Oceanog Microbienne LOMIC, Observ Oceanol Banyuls Mer, F-66650 Banyuls Sur Mer, France. [Carlotti, Francois; Donoso, Katty; Pagano, Marc] Univ Toulon & Var, Aix Marseille Univ, CNRS, IRD,OSU PYTHEAS,MIO,UM110, F-13288 Marseille 09, France. [Escribano, Ruben] Univ Concepcion, Dept Oceanog, Concepcion, Chile. [Escribano, Ruben] Univ Concepcion, Inst Milenio Oceanog, Concepcion, Chile. [Molina, Veronica] Univ Playa Ancha, Dept Biol, Fac Ciencias Nat & Exactas, Valparaiso, Chile. [Molina, Veronica] Univ Playa Ancha, Fac Ciencias Nat & Exactas, Programa Biodiversidad, Valparaiso, Chile. [Fernandez, Camila] Univ Concepcion, FONDAP INCAR Ctr 15110027, Concepcion, Chile. [Fernandez, Camila] Univ Concepcion, Dept Oceanog, COPAS Austral Program, PFB 31, Concepcion, Chile. [Fernandez, Camila] Ctr FONDAP, Invest Dinam Ecosistemas Marinos Altas Latitudes, Valdivia, Chile. RP Valdés, V (corresponding author), Univ Concepcion, Fac Ciencias Nat & Oceanog, Dept Oceanog, Programa Doctorado Oceanog, Concepcion, Chile.; Valdés, V (corresponding author), UPMC Univ Paris 06, Sorbonne Univ, Lab Oceanog Microbienne LOMIC, Observ Oceanol Banyuls Mer, F-66650 Banyuls Sur Mer, France. 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Bakenhus, Insa Balmonte, John Paul Billerbeck, Sara Dlugosch, Leon Henkel, Rohan Kuerzel, Birgit Meyerjuergens, Jens Milke, Felix Voss, Daniela Wienhausen, Gerrit Wietz, Matthias Winkler, Holger Wolterink, Mathias Simon, Meinhard TI Microbial Growth and Organic Matter Cycling in the Pacific Ocean Along a Latitudinal Transect Between Subarctic and Subantarctic Waters SO FRONTIERS IN MARINE SCIENCE DT Article AB The Pacific Ocean constitutes about half of the global oceans and thus microbial processes in this ocean have a large impact on global elemental cycles. Despite several intensely studied regions large areas are still greatly understudied regarding microbial activities, organic matter cycling and biogeography. Refined information about these features is most important to better understand the significance of this ocean for global biogeochemical and elemental cycles. Therefore we investigated a suite of microbial and geochemical variables along a transect from the subantarctic to the subarctic Pacific in the upper 200 m of the water column. The aim was to quantify rates of organic matter processing, identify potential controlling factors and prokaryotic key players. The assessed variables included abundance of heterotrophic prokaryotes and cyanobacteria, heterotrophic prokaryotic production (HPP), turnover rate constants of amino acids, glucose, and acetate, leucine aminopeptidase and beta-glucosidase activities, and the composition of the bacterial community by fluorescence in situ hybridization (FISH). The additional quantification of nitrate, dissolved amino acids and carbohydrates, chlorophyll a, particulate organic carbon and nitrogen (POC, PON) provided a rich environmental context. The oligotrophic gyres exhibited the lowest prokaryotic abundances, rates of HPP and substrate turnover. Low nucleic acid prokaryotes dominated in these gyres, whereas in temperate and subpolar regions further north and south, high nucleic acid prokaryotes dominated. Turnover rate constants of glucose and acetate, as well as leucine aminopeptidase activity, increased from (sub)tropical toward the subpolar regions. In contrast, HPP and bulk growth rates were highest near the equatorial upwelling and lowest in the central gyres and subpolar regions. The SAR11 clade, the Roseobacter group and Flavobacteria constituted the majority of the prokaryotic communities. Vertical profiles of the biogeochemical and microbial variables markedly differed among the different regions and showed close covariations of the microbial variables and chlorophyll a, POC and PON. The results show that hydrographic, microbial, and biogeochemical properties exhibited distinct patterns reflecting the biogeographic provinces along the transect. The microbial variables assessed contribute to a better and refined understanding of the scales of microbial organic matter processing in large areas of the epipelagic Pacific beyond its well-studied regions. C1 [Giebel, Helge-Ansgar; Badewien, Thomas H.; Bakenhus, Insa; Billerbeck, Sara; Dlugosch, Leon; Henkel, Rohan; Kuerzel, Birgit; Meyerjuergens, Jens; Milke, Felix; Voss, Daniela; Wienhausen, Gerrit; Wietz, Matthias; Winkler, Holger; Wolterink, Mathias; Simon, Meinhard] Carl von Ossietzky Univ Oldenburg, Inst Chem & Biol Marine Environm, Oldenburg, Germany. [Arnosti, Carol; Balmonte, John Paul] Univ N Carolina, Dept Marine Sci, Chapel Hill, NC 27515 USA. [Simon, Meinhard] Univ Oldenburg HIFMB, Helmholtz Inst Funct Marine Biodivers, Oldenburg, Germany. [Balmonte, John Paul] Univ Southern Denmark, Dept Biol, HADAL & NordCEE, Odense, Denmark. [Wietz, Matthias] Helmholtz Ctr Polar & Marine Res, Alfred Wegener Inst, Bremerhaven, Germany. RP Simon, M (corresponding author), Carl von Ossietzky Univ Oldenburg, Inst Chem & Biol Marine Environm, Oldenburg, Germany.; Simon, M (corresponding author), Univ Oldenburg HIFMB, Helmholtz Inst Funct Marine Biodivers, Oldenburg, Germany. 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Amin, Shady A. TI Rhodobacteraceae are key players in microbiome assembly of the diatom Asterionellopsis glacialis SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB The complex interactions between bacterioplankton and phytoplankton have prompted numerous studies that investigate phytoplankton microbiomes with the aim of characterizing beneficial or opportunistic taxa and elucidating core bacterial members. Oftentimes, this knowledge is garnered through 16S rRNA gene profiling of microbiomes from phytoplankton isolated across spatial and temporal scales, yet these studies do not offer insight into microbiome assembly and structuring. In this study, we aimed to identify taxa central to structuring and establishing the microbiome of the ubiquitous diatom Asterionellopsis glacialis. We introduced a diverse environmental bacterial community to A. glacialis in nutrient-rich or nutrient-poor media in a continuous dilution culture setup and profiled the bacterial community over 7 days. 16S rRNA amplicon sequencing showed that cyanobacteria (Coleofasciculaceae) and Rhodobacteraceae dominate the microbiome early on and maintain a persistent association throughout the experiment. Differential abundance, co-abundance networks, and differential association analyses revealed that specific members of the family Rhodobacteraceae, particularly Sulfitobacter amplicon sequence variants, become integral members in microbiome assembly. In the presence of the diatom, Sulfitobacter species and other Rhodobacteraceae developed positive associations with taxa that are typically in high abundance in marine ecosystems (Pelagibacter and Synechococcus), leading to restructuring of the microbiome compared to diatom-free controls. These positive associations developed predominantly under oligotrophic conditions, highlighting the importance of investigating phytoplankton microbiomes in as close to natural conditions as possible to avoid biases that develop under routine laboratory conditions. These findings offer further insight into phytoplankton-bacteria interactions and illustrate the importance of Rhodobacteraceae, not merely as phytoplankton symbionts but as key taxa involved in microbiome assembly. IMPORTANCE Most, if not all, microeukaryotic organisms harbor an associated microbial community, termed the microbiome. The microscale interactions that occur between these partners have global-scale consequences, influencing marine primary productivity, carbon cycling, and harmful algal blooms to name but a few. Over the last decade, there has been a growing interest in the study of phytoplankton microbiomes, particularly within the context of bloom dynamics. However, long-standing questions remain regarding the process of phytoplankton microbiome assembly. The significance of our research is to tease apart the mechanism of microbiome assembly with a particular focus on identifying bacterial taxa, which may not merely be symbionts but architects of the phytoplankton microbiome. Our results strengthen the understanding of the ecological mechanisms that underpin phytoplankton-bacteria interactions in order to accurately predict marine ecosystem responses to environmental perturbations. C1 [Isaac, Ashley; Mohamed, Amin R.; Amin, Shady A.] New York Univ Abu Dhabi, Biol Program, Marine Microbi Lab, Abu Dhabi, U Arab Emirates. [Isaac, Ashley] Max Planck Inst Marine Microbiol, Dept Mol Ecol, Bremen, Germany. [Amin, Shady A.] New York Univ Abu Dhabi, Ctr Genom & Syst Biol, Abu Dhabi, U Arab Emirates. [Amin, Shady A.] New York Univ Abu Dhabi, Mubadala ACCESS Ctr, Abu Dhabi, U Arab Emirates. RP Amin, SA (corresponding author), New York Univ Abu Dhabi, Biol Program, Marine Microbi Lab, Abu Dhabi, U Arab Emirates.; Amin, SA (corresponding author), New York Univ Abu Dhabi, Ctr Genom & Syst Biol, Abu Dhabi, U Arab Emirates.; Amin, SA (corresponding author), New York Univ Abu Dhabi, Mubadala ACCESS Ctr, Abu Dhabi, U Arab Emirates. 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Wiltshire, Karen H. Schweder, Thomas Fuchs, Bernhard M. Bengtsson, Mia M. Teeling, Hanno Amann, Rudolf I. TI Particle-attached bacteria act as gatekeepers in the decomposition of complex phytoplankton polysaccharides SO MICROBIOME DT Article AB Background Marine microalgae (phytoplankton) mediate almost half of the worldwide photosynthetic carbon dioxide fixation and therefore play a pivotal role in global carbon cycling, most prominently during massive phytoplankton blooms. Phytoplankton biomass consists of considerable proportions of polysaccharides, substantial parts of which are rapidly remineralized by heterotrophic bacteria. We analyzed the diversity, activity, and functional potential of such polysaccharide-degrading bacteria in different size fractions during a diverse spring phytoplankton bloom at Helgoland Roads (southern North Sea) at high temporal resolution using microscopic, physicochemical, biodiversity, metagenome, and metaproteome analyses. Results Prominent active 0.2-3 mu m free-living clades comprised Aurantivirga, "Formosa", Cd. Prosiliicoccus, NS4, NS5, Amylibacter, Planktomarina, SAR11 Ia, SAR92, and SAR86, whereas BD1-7, Stappiaceae, Nitrincolaceae, Methylophagaceae, Sulfitobacter, NS9, Polaribacter, Lentimonas, CL500-3, Algibacter, and Glaciecola dominated 3-10 mu m and > 10 mu m particles. Particle-attached bacteria were more diverse and exhibited more dynamic adaptive shifts over time in terms of taxonomic composition and repertoires of encoded polysaccharide-targeting enzymes. In total, 305 species-level metagenome-assembled genomes were obtained, including 152 particle-attached bacteria, 100 of which were novel for the sampling site with 76 representing new species. Compared to free-living bacteria, they featured on average larger metagenome-assembled genomes with higher proportions of polysaccharide utilization loci. The latter were predicted to target a broader spectrum of polysaccharide substrates, ranging from readily soluble, simple structured storage polysaccharides (e.g., laminarin, alpha-glucans) to less soluble, complex structural, or secreted polysaccharides (e.g., xylans, cellulose, pectins). In particular, the potential to target poorly soluble or complex polysaccharides was more widespread among abundant and active particle-attached bacteria. Conclusions Particle-attached bacteria represented only 1% of all bloom-associated bacteria, yet our data suggest that many abundant active clades played a pivotal gatekeeping role in the solubilization and subsequent degradation of numerous important classes of algal glycans. The high diversity of polysaccharide niches among the most active particle-attached clades therefore is a determining factor for the proportion of algal polysaccharides that can be rapidly remineralized during generally short-lived phytoplankton bloom events. C1 [Wang, Feng-Qing; Sidhu, Chandni; Lu, De-Chen; Fuchs, Bernhard M.; Teeling, Hanno; Amann, Rudolf I.] Max Planck Inst Marine Microbiol, Celsiusstr 1, D-28359 Bremen, Germany. [Bartosik, Daniel; Schweder, Thomas] Univ Greifswald, Inst Pharm, Felix Hausdorff Str 3, D-17489 Greifswald, Germany. [Bartosik, Daniel; Schweder, Thomas] Inst Marine Biotechnol, Walther Rathenau Str 49a, D-17489 Greifswald, Germany. [Siebers, Robin; Trautwein-Schult, Anke; Becher, Doerte; Bengtsson, Mia M.] Univ Greifswald, Inst Microbiol, Felix Hausdorff Str 8, D-17489 Greifswald, Germany. [Lu, De-Chen] Shandong Univ, Marine Coll, Weihai 264209, Peoples R China. [Huettel, Bruno] Max Planck Genome Ctr Cologne, Carl von Linne Weg 10, D-50829 Cologne, Germany. [Rick, Johannes; Kirstein, Inga V.; Wiltshire, Karen H.] Alfred Wegener Inst Polar & Marine Res, Biol Anstalt Helgoland, D-27483 Helgoland, Germany. 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Levi, Noah J. Resley, Matthew Main, Kevan TI Effect of copper sulfate on the external microbiota of adult common snook (Centropomus undecimalis) SO ANIMAL MICROBIOME DT Article AB Background The environment exerts a strong influence on the fish external microbiota, with lower diversity and increased abundances of opportunistic bacterial groups characterizing cultured fish compared to their wild counterparts. Deviation from a healthy external microbiota structure has been associated with increased susceptibility to bacterial pathogens. Treatment of wild-caught broodstock with copper sulfate for the removal of external parasites is a common aquaculture practice. Despite the microbiota's importance to fish health, the effects of copper sulfate on mucosal bacterial communities and their ability to recover following this chemical treatment have not been examined. The skin microbiota of adult common snook was characterized from wild individuals (Wild), and wild-caught fish maintained in recirculating aquaculture systems (RAS) immediately following a month-long copper sulfate treatment (Captive-1), and then two-weeks (Captive-2) and 2 years (Captive-3) after cessation of copper treatment. Results The skin microbiota of wild fish were characterized by high diversity and taxa including Synechocococcus, SAR11, and a member of the Roseobacter clade. Bacterial diversity decreased in Captive individuals during the 2-year sampling period. Captive fish harbored greater abundances of Firmicutes, which may reflect glycan differences between aquaculture and natural feeds. Bacterial taxa with copper resistance mechanisms and indicative of metal contamination were enriched in Captive-1 and Captive-2 fish. Vibrionaceae were dominant in Captive fish, particularly immediately and 2 weeks following copper treatment. Based on our observations and previous literature, our results suggest putatively beneficial taxa amass over time in captivity. Within 2 years, Captive individuals harbored Bacillus which contains numerous probiotic candidates and the complex carbon degraders of the family Saprospiraceae. Predicted butanoate metabolism exceeded that of Wild fish, and its reported roles in immunity and energy provision suggest a prebiotic effect for fishes. Conclusions The mucosal microbiota contains bacterial taxa that may act as bioindicators of environmental pollution. Increases in mutualistic groups indicate a return to a beneficial skin microbiota following copper sulfate treatment. Our data also suggests that vastly different taxa, influenced by environmental conditions, can be associated with adult fish without noticeable health impairment, perhaps due to establishment of various mutualists to maintain fish mucosal health. C1 [Tarnecki, Andrea M.] Mote Marine Lab, Marine Immunol Program, 1600 Ken Thompson Pkwy, Sarasota, FL 34236 USA. [Levi, Noah J.] Wabash Coll, Dept Biol, 301 West Wabash Ave, Crawfordsville, IN 47933 USA. [Levi, Noah J.] Univ Miami, Med Scientist Training Program, Miller Sch Med, 1600 NW 10th Ave, Miami, FL 33101 USA. [Resley, Matthew; Main, Kevan] Mote Aquaculture Res Pk, Directorate Fisheries & Aquaculture, 874 WR Mote Way, Sarasota, FL 34240 USA. RP Tarnecki, AM (corresponding author), Mote Marine Lab, Marine Immunol Program, 1600 Ken Thompson Pkwy, Sarasota, FL 34236 USA. 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Polaribacter belonging to Bacteroidetes was found to be dominant in the peak phase, and its transcriptional activity was high (48.9% of the total mRNA reads). Two dominant Polaribacter bins had the potential to utilize major polymers in P. antarctica, chrysolaminarin and xylan, with a distinct set of glycosyl hydrolases. In the decline phase, Gammaproteobacteria (Ant4D3, SUP05, and SAR92), with the potential to utilize low molecular weight-dissolved organic matter (LMW-DOM) including compatible solutes, was increased. The versatility of Gammaproteobacteria may contribute to their abundance in organic carbon-rich polynya waters, while the SAR11 clade was found to be predominant in the sea ice-covered oligotrophic ocean. SAR92 clade showed transcriptional activity for utilization of both polysaccharides and LMW-DOM; this may account for their abundance both in the peak and decline phases. Ant4D3 clade was dominant in all phases of the polynya bloom, implicating the crucial roles of this clade in LMW-DOM remineralization in the Antarctic polynyas. Genomic reconstruction and in situ gene expression analyses revealed the unique metabolic potential of dominant bacteria of the Antarctic polynya at a finer taxonomic level. The information can be used to predict temporal community succession linked to the availability of substrates derived from the P. antarctica bloom. Global warming has resulted in compositional changes in phytoplankton from P. antarctica to diatoms, and thus, repeated parallel studies in various polynyas are required to predict global warming-related changes in carbon remineralization. C1 [Kim, So-Jeong] Korea Inst Geosci & Mineral Resources, Geol Environm Res Div, Daejeon 34132, South Korea. [Kim, Jong-Geol; Gwak, Joo-Han; Rhee, Sung-Keun] Chungbuk Natl Univ, Dept Microbiol, Cheongju 28644, South Korea. [Lee, Sang-Hoon; Yang, Eun-Jin; Park, Jisoo; Jung, Jinyoung] Korea Polar Res Inst, Div Polar Ocean Environm, Incheon 21990, South Korea. [Park, Soo-Je] Jeju Natl Univ, Dept Biol, Jeju 63243, South Korea. [Jung, Man-Young] Univ Vienna, Dept Microbial Ecol, A-1090 Vienna, Austria. [Chung, Won-Hyung] Korea Food Res Inst, Res Grp Gut Microbiome, Sungnam 13539, South Korea. [Hahn, Yoonsoo] Chung Ang Univ, Dept Life Sci, Seoul 06974, South Korea. [Cho, Jang-Cheon] Inha Univ, Dept Biol Sci, Incheon 22212, South Korea. [Madsen, Eugene L.] Cornell Univ, Dept Microbiol, Ithaca, NY 14853 USA. [Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Div Microbiol, Evolutionary Genom Grp, Apartado 18, Alicante, Spain. [Hyun, Jung-Ho] Hanyang Univ, Dept Marine Sci & Convergence Engn, ERICA Campus, Ansan 15588, South Korea. RP Rhee, SK (corresponding author), Chungbuk Natl Univ, Dept Microbiol, Cheongju 28644, South Korea. 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It is essential to understand the structure and metabolic function of microbial communities. The Indian Ocean is the third largest ocean in the world, and it possesses unique hydrographical properties. So far, assessments of microbial diversity and metabolism need to be improved in the Indian Ocean. Therefore, we carried out a series of investigations in the equatorial eastern Indian Ocean in order to clarify the local microbial communities and detect the genetic potential for microbial functions. The obtained results suggested Cyanobacteria was the dominant microbial group, and predicted the Calvin cycle and the assimilatory nitrate and nitrite reduction played important role in the pathway of carbon fixation and nitrogen metabolism respectively. This study provides insights into microbial community structures as well as the metabolic potential that may be active in the local environment, and lays the groundwork for understanding the roles of microbes in energy and resource cycling in this habitat. Currently, there is scant information about the biodiversity and functional diversity of microbes in the eastern Indian Ocean (EIO). Here, we used a combination of high-throughput sequencing of 16S rRNA genes and a metagenomic approach to investigate the microbial population structure and its metabolic function in the equatorial EIO. Our results show that Cyanobacterial Prochlorococcus made up the majority of the population. Interestingly, there were fewer contributions from clades SAR11 (Alphaproteobacteria) and SAR86 (Gammaproteobacteria) to microbial communities than contributions from Prochlorococcus. Based on functional gene analysis, functional genes rbcL, narB, and nasA were relatively abundant among the relevant genes. 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These studies, however, have shown that at least some of the "alphas" differ fundamentally in the ways in which they regulate their genes in response to Fe availability. In this paper, we build on our own work on Rhizobium leguminosarum (the symbiont of peas, beans and clovers) and on Bradyrhizobium japonicum, which nodulates soybeans and which has been studied in Buffalo and Zurich. In the former species, the predominant Fe-responsive regulator is not Fur, but RirA, a member of the Rrf2 protein family and which likely has an FeS cluster cofactor. In addition, there are several R. leguminosarum genes that are expressed at higher levels in Fe-replete conditions and at least some of these are regulated by Irr, a member of the Fur superfamily and which has the unusual property of being degraded by the presence of heme. In silico analyses of the genome sequences of other bacteria indicate that Irr occurs in all members of the Rhizobiales and the Rhodobacterales and that RirA is found in all but one branch of these two lineages, the exception being the clade that includes B. japonicum. Nearly all the Rhizobiales and the Rhodobacterales contain a gene whose product resembles bona fide Fur. However, direct genetic studies show that in most of the Rhizobiales and in the Rhodobacterales it is a "Mur" (a manganese responsive repressor of a small number of genes involved in Mn uptake) or, in Bradyrhizobium, it recognizes the operator sequences of only a few genes that are involved in Fe metabolism. We propose that the Rhizobiales and the Rhodobacterales have relegated Fur to a far more minor role than in (say) E. coli and that they employ Irr and, in the Rhizobiales, RirA as their global Fe-responsive transcriptional regulators. In contrast to the direct interaction between Fe2+ and conventional Fur, we suggest that these bacteria sense Fe more indirectly as functions of the intracellular concentrations of FeS clusters and of heme. Thus, their "iron-omes" may be more accurately linked to the real-time needs for the metal and not just to its absolute concentration in the environment. C1 Univ E Anglia, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England. Russian Acad Sci, Inst Informat Transmiss Problems, Moscow 127994, Russia. Moscow MV Lomonosov State Univ, Dept Bioengn & Bioinformat, Moscow 119992, Russia. Burnham Inst Med Res, La Jolla, CA 92037 USA. RP Johnston, AWB (corresponding author), Univ E Anglia, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England. 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Alarcon-Schumacher, Tomas Tamayo-Leiva, Javier Pedros-Alio, Carlos Farias, Laura Diez, Beatriz TI Proteorhodopsin Phototrophy in Antarctic Coastal Waters SO MSPHERE DT Article AB Microbial proton-pumping rhodopsins are considered the simplest strategy among phototrophs to conserve energy from light. Proteorhodopsins are the most studied rhodopsins thus far because of their ubiquitous presence in the ocean, except in Antarctica, where they remain understudied. We analyzed proteorhodopsin abundance and transcriptional activity in the Western Antarctic coastal seawaters. Combining quantitative PCR (qPCR) and metagenomics, the relative abundance of proteorhodopsin-bearing bacteria accounted on average for 17, 3.5, and 29.7% of the bacterial community in Chile Bay (South Shetland Islands) during 2014, 2016, and 2017 summer-autumn, respectively. The abundance of proteorhodopsin-bearing bacteria changed in relation to environmental conditions such as chlorophyll a and temperature. Alphaproteobacteria, Gammaproteobacteria, and Flavobacteriia were the main bacteria that transcribed the proteorhodopsin gene during day and night. Although green light-absorbing proteorhodopsin genes were more abundant than blue-absorbing ones, the latter were transcribed more intensely, resulting in .50% of the proteorhodopsin transcripts during the day and night. Flavobacteriia were the most abundant proteorhodopsin-bearing bacteria in the metagenomes; however, Alphaproteobacteria and Gammaproteobacteria were more represented in the metatranscriptomes, with qPCR quantification suggesting the dominance of the active SAR11 clade. Our results show that proteorhodopsin-bearing bacteria are prevalent in Antarctic coastal waters in late austral summer and early autumn, and their ecological relevance needs to be elucidated to better understand how sunlight energy is used in this marine ecosystem. IMPORTANCE Proteorhodopsin-bearing microorganisms in the Southern Ocean have been overlooked since their discovery in 2000. The present study identify taxonomy and quantify the relative abundance of proteorhodopsin-bearing bacteria and proteorhodopsin gene transcription in the West Antarctic Peninsula's coastal waters. This information is crucial to understand better how sunlight enters this marine environment through alternative ways unrelated to chlorophyll-based strategies. The relative abundance of proteorhodopsin-bearing bacteria seems to be related to environmental parameters (e.g., chlorophyll a, temperature) that change yearly at the coastal water of the West Antarctic Peninsula during the austral late summers and early autumns. Proteorhodopsin-bearing bacteria from Antarctic coastal waters are potentially able to exploit both the green and blue spectrum of sunlight and are a prevalent group during the summer in this polar environment. 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Denitrification genes were enriched in >30 mu m particles, even in the oxycline, while anammox bacteria were not abundant on particles. Many steps in denitrification were encoded by multiple phylotypes with different distributions. Notably three N2O reductases (nosZ), each with no cultured relative, inhabited distinct niches; one was free-living, one dominant on particles and one had a C terminal extension found in autotrophic S-oxidizing bacteria. At some depths > 30% of the community possessed nitrite reductase nirK. A nirK OTU linked to SAR11 explained much of this abundance. The only bacterial gene found for NO reduction to N2O in the ODZ was a form of qnor B related to the previously postulated "nitric oxide dismutase," hypothesized to produce N-2 directly while oxidizing methane. 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V. Goodwin, K. D. TI Long-Read Sequencing Improves Recovery of Picoeukaryotic Genomes and Zooplankton Marker Genes from Marine Metagenomes SO MSYSTEMS DT Article AB Ocean microbes provide critical ecosystem services, but most remain uncultivated. Their communities can be studied through shotgun metagenomic sequencing and bioinformatic analyses, including binning draft microbial genomes. Long-read sequencing offers the potential to improve metagenome assemblies and provide more robust assessments of microbial community composition and function than short-read sequencing. We applied Pacific Biosciences (PacBio) CCS (circular consensus sequencing) HiFi shotgun sequencing to 14 marine water column samples and compared the results with those for short-read metagenomes from the corresponding environmental DNA samples. We found that long-read metagenomes varied widely in quality and biological information. The community compositions of the corresponding long- and short-read metagenomes were frequently dissimilar, suggesting higher stochasticity and/or bias associated with PacBio sequencing. Long reads provided few improvements to the assembly qualities, gene annotations, and prokaryotic metagenome-assembled genome (MAG) binning results. However, only long reads produced high-quality eukaryotic MAGs and contigs containing complete zooplankton marker gene sequences. These results suggest that high-quality long-read metagenomes can improve marine community composition analyses and provide important insight into eukaryotic phyto- and zooplankton genetics, but the benefits may be outweighed by the inconsistent data quality.IMPORTANCE Ocean microbes provide critical ecosystem services, but most remain uncultivated. Their communities can be studied through shotgun metagenomic sequencing and bioinformatic analyses, including binning draft microbial genomes. However, most sequencing to date has been done using short-read technology, which rarely yields genome sequences of key microbes like SAR11. Long-read sequencing can improve metagenome assemblies but is hampered by technological shortcomings and high costs. In this study, we compared long- and short-read sequencing of marine metagenomes. We found a wide range of long-read metagenome qualities and minimal improvements to microbiome analyses. However, long reads generated draft genomes of eukaryotic algal species and provided full-length marker gene sequences of zooplankton species, including krill and copepods. These results suggest that long-read sequencing can provide greater genetic insight into the wide diversity of eukaryotic phyto- and zooplankton that interact as part of and with the marine microbiome. C1 [Patin, N. V.; Goodwin, K. D.] NOAA, Ocean Chem & Ecosyst Div, Atlantic Oceanog & Meteorol Lab, Miami, FL 33149 USA. [Patin, N. 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Coutinho, Felipe H. Gregoracci, Gustavo B. Leomil, Luciana de Oliveira, Louisi S. Froes, Adriana Tschoeke, Diogo Soares, Ana Carolina Cabral, Anderson S. Ward, Nicholas D. Richey, Jeffrey E. Krusche, Alex V. Yager, Patricia L. de Rezende, Carlos Eduardo Thompson, Cristiane C. Thompson, Fabiano L. TI Virioplankton Assemblage Structure in the Lower River and Ocean Continuum of the Amazon SO MSPHERE DT Article AB The Amazon River watershed and its associated plume comprise a vast continental and oceanic area. The microbial activities along this continuum contribute substantially to global carbon and nutrient cycling, and yet there is a dearth of information on the diversity, abundance, and possible roles of viruses in this globally important river. The aim of this study was to elucidate the diversity and structure of virus assemblages of the Amazon River-ocean continuum. Environmental viral DNA sequences were obtained for 12 locations along the river's lower reach (n = 5) and plume (n = 7). Sequence assembly yielded 29,358 scaffolds, encoding 82,546 viral proteins, with 15 new complete viral genomes. Despite the spatial connectivity mediated by the river, virome analyses and physical-chemical water parameters clearly distinguished river and plume ecosystems. Bacteriophages were ubiquitous in the continuum and were more abundant in the transition region. Eukaryotic viruses occurred mostly in the river, while the plume had more viruses of autotrophic organisms (Prochlorococcus, Synechococcus) and heterotrophic bacteria (Pelagibacter). The viral families Microviridae and Myoviridae were the most abundant and occurred throughout the continuum. The major functions of the genes in the continuum involved viral structures and life cycles, and viruses from plume locations and Tapajos River showed the highest levels of functional diversity. The distribution patterns of the viral assemblages were defined not only by the occurrence of possible hosts but also by water physical and chemical parameters, especially salinity. The findings presented here help to improve understanding of the possible roles of viruses in the organic matter cycle along the river-ocean continuum. IMPORTANCE The Amazon River forms a vast plume in the Atlantic Ocean that can extend for more than 1,000 km. Microbial communities promote a globally relevant carbon sink system in the plume. Despite the importance of viruses for the global carbon cycle, the diversity and the possible roles of viruses in the Amazon are poorly understood. The present work assesses, for the first time, the abundance and diversity of viruses simultaneously in the river and ocean in order to elucidate their possible roles. DNA sequence assembly yielded 29,358 scaffolds, encoding 82,546 viral proteins, with 15 new complete viral genomes from the 12 river and ocean locations. 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This study investigated the marine prokaryotic communities across 22 stations spanning the Arctic Ocean, the Chukchi Sea, the Bering Sea, and the Sea of Japan, with an emphasis on how environmental factors shape these communities. Results showed that the microbial alpha diversity generally declines with increasing latitude, though Arctic Ocean stations exhibited higher Chao 1 indices compared to the Bering Sea. Beta diversity analyses revealed that temperature and salinity were key factors associated with community composition variation across latitudes. Proteobacteria and Cyanobacteria were the dominant phyla showing opposite distribution trends across sampling stations. Cold-adapted oligotrophs such as Planktomarina and the SAR11 clade thrived in Arctic waters, while Sphingomonas, known for pollutant degradation, was more abundant in the Sea of Japan. Temperature was positively correlated to the relative abundance of Sphingomonas. At broad spatial scales, stochastic processes dominated community assembly of microbial phylogenetic diversity, while in specific regions like the Arctic Ocean, deterministic homogeneous selection appeared to shape microbial communities; and temperature showed a pronounced influence on phylogenetic turnover across all samples. Co-occurrence networks identified several key taxa, such as Polaribacter_1, Candidatus_Aquiluna, and NS5_marine_group. Overall, the study underscores temperature's role in shaping microbial community diversity, composition, and assembly processes across latitudinal gradients, highlighting unique community adaptations to extreme environments.IMPORTANCEMicrobes are the invisible engines of ocean health, recycling nutrients and sustaining marine life. This research helps us understand how climate factors like temperature shape these microscopic communities, which differ starkly between icy Arctic waters and warmer seas. As oceans warm due to climate change, microbial populations and their critical roles in cleaning pollutants or supporting food webs could shift dramatically. The study suggests Arctic microbes are uniquely adapted to cold, low-nutrient conditions, making them vulnerable to warming. By linking temperature to microbial diversity, this work provides clues to predict how marine ecosystems might respond to climate shifts, informing efforts to protect ocean biodiversity and processes vital to Earth's carbon and nutrient cycles. C1 [Pan, Ying; Tao, Ye; Yang, Xian; Du, Siyi; Ding, Hongguang; Li, Jiaxin; Jia, Hanwen; Chen, Huaihai] Sun Yat Sen Univ, Sch Ecol, State Key Lab Biocontrol, Shenzhen Campus, Shenzhen, Guangdong, Peoples R China. RP Chen, HH (corresponding author), Sun Yat Sen Univ, Sch Ecol, State Key Lab Biocontrol, Shenzhen Campus, Shenzhen, Guangdong, Peoples R China. 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However, comprehensive evaluations of viral infections in natural prokaryotic communities are still technically difficult. Viruses infecting marine prokaryotes have a large impact on the diversity and dynamics of their hosts. Model systems suggest that viral infection is frequency dependent and constrained by the virus-host encounter rate. However, it is unclear whether frequency-dependent infection is pervasive among the abundant prokaryotic populations with different temporal dynamics. To address this question, we performed a comparison of prokaryotic and viral communities using 16S rRNA amplicon and virome sequencing based on samples collected monthly for 2 years at a Japanese coastal site, Osaka Bay. Concurrent seasonal shifts observed in prokaryotic and viral community dynamics indicated that the abundance of viruses correlated with that of their predicted host phyla (or classes). 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An imbalance between high demand and limited production, in concert with abiotic processes, may explain the low availability of these vitamins in marine systems. Natural microbial communities from surface shelf water in the productive area off northwestern Spain were enclosed in mesocosms in winter, spring, and summer 2016. In order to explore the impact of B-vitamin availability on microbial community composition (16S and 18S rRNA gene sequence analysis) and bacterial function (metatranscriptomics analysis) in different seasons, enrichment experiments were conducted with seawater from the mesocosms. Our findings revealed that significant increases in phytoplankton or prokaryote biomass associated with vitamin B-12 and/or B-1 amendments were not accompanied by significant changes in community composition, suggesting that most of the microbial taxa benefited from the external B-vitamin supply. Metatranscriptome analysis suggested that many bacteria were potential consumers of vitamins B-12 and B-1, although the relative abundance of reads related to synthesis was ca. 3.6-fold higher than that related to uptake. Alteromonadales and Oceanospirillales accounted for important portions of vitamin B-1 and B-12 synthesis gene transcription, despite accounting for only minor portions of the bacterial community. Flavobacteriales appeared to be involved mostly in vitamin B-12 and B-1 uptake, and Pelagibacterales expressed genes involved in vitamin B-1 uptake. Interestingly, the relative expression of vitamin B-12 and B-1 synthesis genes among bacteria strongly increased upon inorganic nutrient amendment. Collectively, these findings suggest that upwelling events intermittently occurring during spring and summer in productive ecosystems may ensure an adequate production of these cofactors to sustain high levels of phytoplankton growth and biomass. IMPORTANCE B vitamins are essential growth factors for practically all living organisms on Earth that are produced by a selection of microorganisms. An imbalance between high demand and limited production may explain the low concentration of these compounds in marine systems. In order to explore the impact of B-vitamin availability on bacteria and algae in the coastal waters off northwestern Spain, six experiments were conducted with natural surface water enclosed in winter, spring, and summer. Our findings revealed that increases in phytoplankton or bacterial growth associated with B-12 and/or B-1 amendments were not accompanied by significant changes in community composition, suggesting that most microorganisms benefited from the B-vitamin supply. Our analyses confirmed the role of many bacteria as consumers of vitamins B-12 and B-1, although the relative abundance of genes related to synthesis was ca. 3.6-fold higher than that related to uptake. Interestingly, prokaryote expression of B-12 and B-1 synthesis genes strongly increased when inorganic nutrients were added. Collectively, these findings suggest that upwelling of cold and nutrient-rich waters occurring during spring and summer in this coastal area may ensure an adequate production of B vitamins to sustain high levels of algae growth and biomass. C1 [Joglar, Vanessa; Martinez-Garcia, Sandra; Fuentes-Lema, Antonio; Perez-Lorenzo, Maria; Fernandez, Emilio; Teira, Eva] Univ Vigo, Ctr Invest Marina Univ Vigo, Dept Ecol & Biol Anim, Vigo, Spain. [Pontiller, Benjamin; Lundin, Daniel; Pinhassi, Jarone] Linnaeus Univ, Ctr Ecol & Evolut Microbial Model Syst, Kalmar, Sweden. [Pontiller, Benjamin] GEOMAR Helmholtz Ctr Ocean Res, Kiel, Germany. RP Joglar, V (corresponding author), Univ Vigo, Ctr Invest Marina Univ Vigo, Dept Ecol & Biol Anim, Vigo, Spain. 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Piwosz, Kasia Santic, Danijela Sestanovic, Stefanija Tomas, Ana Vrdoljak Hanusova, Martina Solic, Mladen Koblizek, Michal TI Lineage-Specific Growth Curves Document Large Differences in Response of Individual Groups of Marine Bacteria to the Top-Down and Bottom-Up Controls SO MSYSTEMS DT Article AB Marine bacterioplankton represent a diverse assembly of species differing largely in their abundance, physiology, metabolic activity, and role in microbial food webs. To analyze their sensitivity to bottom-up and top-down controls, we performed a manipulation experiment where grazers were removed, with or without the addition of phosphate. Using amplicon-reads normalization by internal standard (ARNIS), we reconstructed growth curves for almost 300 individual phylotypes. Grazer removal caused a rapid growth of most bacterial groups, which grew at rates of 0.6 to 3.5 day(-1), with the highest rates (>4 day(-1)) recorded among Rhodobacteraceae, Oceanospirillales, Alteromonadaceae, and Arcobacteraceae. Based on their growth response, the phylotypes were divided into three basic groups. Most of the phylotypes responded positively to both grazer removal as well as phosphate addition. The second group (containing, e.g., Rhodobacterales and Rhizobiales) responded to the grazer removal but not to the phosphate addition. Finally, some clades, such as SAR11 and Flavobacteriaceae, responded only to phosphate amendment but not to grazer removal. Our results show large differences in bacterial responses to experimental manipulations at the phylotype level and document different life strategies of marine bacterioplankton. In addition, growth curves of 130 phylogroups of aerobic anoxygenic phototrophs were reconstructed based on changes of the functional pufM gene. The use of functional genes together with rRNA genes may significantly expand the scientific potential of the ARNIS technique. IMPORTANCE Growth is one of the main manifestations of life. It is assumed generally that bacterial growth is constrained mostly by nutrient availability (bottom-up control) and grazing (top-down control). Since marine bacteria represent a very diverse assembly of species with different metabolic properties, their growth characteristics also largely differ accordingly. Currently, the growth of marine microorganisms is typically evaluated using microscopy in combination with fluorescence in situ hybridization (FISH). However, these laborious techniques are limited in their throughput and taxonomical resolution. Therefore, we combined a classical manipulation experiment with next-generation sequencing to resolve the growth dynamics of almost 300 bacterial phylogroups in the coastal Adriatic Sea. 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This study first evaluated the impact of perturbations caused by sequential protozoan blooms of the heterotrophic dinoflagellate Noctiluca scintillans and the mixotrophic ciliate Mesodinium rubrum on the structuring of these two bacterial communities. During a series of blooms of Noctiluca scintillans and Mesodinium rubrum, we applied high-throughput sequencing of the 16S rRNA gene to investigate the population dynamics of free-living (FL) and particle-attached (PA) bacteria in an attempt to evaluate the influence of protozoan bloom-induced disturbances on the structuring of these two communities. Our findings revealed that the FL and PA bacterial community compositions (BCCs) displayed distinct profiles during sequential blooms, and the PA flora responded more dynamically to these pulse perturbations. The dominant bacterial groups (e.g., Flavobacteriaceae, Rhodobacteraceae, Vibrionaceae, and SAR11 subclade I) in these two communities displayed different levels of connectivity with the bloom-causative species and environmental factors. In addition, more FL bacterial groups were associated with M. rubrum, while more PA bacterial groups were related to N. scintillans. Potential endocytic bacteria of N. scintillans, particularly Vibrionaceae and Rickettsiaceae, opportunistically thrived at the peak of the bloom, suggesting that they could be important players influencing the dynamics and biogeochemical cycling of the blooms. Overall, disparities in the substrate preferences and thermal niches of various bacterial taxa as well as the short duration of the blooms (1 to 3 days) contributed to the diverse responses of the FL and PA bacterial communities to these protozoan blooms. Our research provides insight into the responses of FL and PA bacterial communities to blooms caused by protozoa like N. scintillans and M. rubrum and highlights the ecological significance of certain keystone bacterial groups during this kind of cosmopolitan protozoan bloom. IMPORTANCE Shifts in the bacterioplankton community composition during phytoplankton blooms have been studied extensively; however, investigations on protozoan blooms are rare. This study first evaluated the impact of perturbations caused by sequential protozoan blooms of the heterotrophic dinoflagellate Noctiluca scintillans and the mixotrophic ciliate Mesodinium rubrum on the structuring of these two bacterial communities. Our findings shed light on the responses of these two bacterial communities to such cosmopolitan protozoan blooms and highlight the possible ecological significance of certain keystone bacterial groups during these blooms. This research prepares the way for more focused studies that will help in understanding the roles that bacteria play during protozoan blooms and their impact on environmental health. C1 [Zhang, Shuwen] South China Normal Univ, Sch Life Sci, Guangzhou Key Lab Subtrop Biodivers & Biomonitori, Guangdong Prov Key Lab Hlth & Safe Aquaculture, Guangzhou, Peoples R China. [Liu, Hongbin] Hong Kong Univ Sci & Technol, Div Life Sci, Clear Water Bay, Hong Kong, Peoples R China. [Liu, Hongbin] Hong Kong Univ Sci & Technol, Dept Ocean Sci, Clear Water Bay, Hong Kong, Peoples R China. [Xia, Xiaomin] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangdong Prov Key Lab Appl Marine Biol, Guangzhou, Peoples R China. RP Zhang, SW (corresponding author), South China Normal Univ, Sch Life Sci, Guangzhou Key Lab Subtrop Biodivers & Biomonitori, Guangdong Prov Key Lab Hlth & Safe Aquaculture, Guangzhou, Peoples R China.; Xia, XM (corresponding author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangdong Prov Key Lab Appl Marine Biol, Guangzhou, Peoples R China. 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Eppley, John M. Romano, Anna E. Luo, Elaine DeLong, Edward F. Karl, David M. TI Microbial Sources of Exocellular DNA in the Ocean SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB With advances in metagenomic sequencing, the microbial composition of diverse environmental systems has been investigated, providing new perspectives on potential ecological dynamics and dimensions for experimental investigations. Here, we characterized exocellular free DNA via metagenomics, using a newly developed method that separates free DNA from cells, viruses, and vesicles, and facilitated the independent characterization of each fraction. Exocellular DNA is operationally defined as the fraction of the total DNA pool that passes through a membrane filter (0.1 mu m). It is composed of DNA-containing vesicles, viruses, and free DNA and is ubiquitous in all aquatic systems, although the sources, sinks, and ecological consequences are largely unknown. Using a method that provides separation of these three fractions, we compared open ocean depth profiles of DNA associated with each fraction. Pelagibacter-like DNA dominated the vesicle fractions for all samples examined over a depth range of 75 to 500 m. Viral DNA consisted predominantly of myovirus-like and podovirus-like DNA and contained the highest proportion of unannotated sequences. Euphotic zone free DNA (75 to 125 m) contained primarily bacterial and viral sequences, with bacteria dominating samples from the mesopelagic zone (500 to 1,000 m). A high proportion of mesopelagic zone free DNA sequences appeared to originate from surface waters, including a large amount of DNA contributed by high-light Prochlorococcus ecotypes. Throughout the water column, but especially in the mesopelagic zone, the composition of free DNA sequences was not always reflective of cooccurring microbial communities that inhabit the same sampling depth. These results reveal the composition of free DNA in different regions of the water column (euphotic and mesopelagic zones), with implications for dissolved organic matter cycling and export (by way of sinking particles and/or migratory zooplankton) as a delivery mechanism. IMPORTANCE With advances in metagenomic sequencing, the microbial composition of diverse environmental systems has been investigated, providing new perspectives on potential ecological dynamics and dimensions for experimental investigations. Here, we characterized exocellular free DNA via metagenomics, using a newly developed method that separates free DNA from cells, viruses, and vesicles, and facilitated the independent characterization of each fraction. The fate of this free DNA has both ecological consequences as a nutrient (N and P) source and potential evolutionary consequences as a source of genetic transformation. Here, we document different microbial sources of free DNA at the surface (0 to 200 m) versus depths of 250 to 1,000 m, suggesting that distinct free DNA production mechanisms may be present throughout the oligotrophic water column. Examining microbial processes through the lens of exocellular DNA provides insights into the production of labile dissolved organic matter (i.e., free DNA) at the surface (likely by viral lysis) and processes that influence the fate of sinking, surface-derived organic matter. C1 [Linney, Morgan D.; Eppley, John M.; Romano, Anna E.; Luo, Elaine; DeLong, Edward F.; Karl, David M.] Univ Hawaii Manoa, Daniel K Inouye Ctr Microbial Oceanog Res & Educ, Dept Oceanog, Honolulu, HI 96822 USA. [Linney, Morgan D.] MIT, Dept Civil & Environm Engn, 77 Massachusetts Ave, Cambridge, MA 02139 USA. [Luo, Elaine] Woods Hole Oceanog Inst, Woods Hole, MA 02543 USA. 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10.1128/AEM.02807-16 Zhong X., 2016, AM GEOPH UN OC SCI M NR 70 TC 7 Z9 7 PD APR 12 PY 2022 VL 88 IS 7 AR e02093-21 DI 10.1128/aem.02093-21 UT WOS:000782461400034 DA 2025-07-30 ER PT B AU Okafor, N AF Okafor, Nduka BA Okafor, N BF Okafor, N TI Ecology of Microorganisms in Saline Waters (Seas and Oceans) SO ENVIRONMENTAL MICROBIOLOGY OF AQUATIC AND WASTE SYSTEMS DT Article; Book Chapter AB The waters of the seas and oceans of the world contain large amounts of solutes; mainly, salt of about 3.5 g/L, occupy about 71% of the earth's surface and have an average depth of 3.8 km. The photic zone of seas and oceans, about 200 m deep, is the region permeated by sunlight where photosynthesis can take place. It has the greatest biodiversity, and all food for the marine population arises from the photic zone; such food includes marine snow which consists of globules of mucopolyssacharides containing dead and living microorganisms floating downward toward the deep ocean. Marine organisms are adapted to the unique conditions found in the marine open sea (pelagic zone) environment: high salinity (3.5 g/L), low temperature (about 4 degrees C), and high barometric pressure of up to 500 bar depending on the depth. Thermophilic organisms grow near the occasional hot thermal vents where hot magma spews out onto the ocean floor. Using the technique of 16S rRNA, it has been found that over 70% of marine bacteria have not been cultured and hence have no counterparts among known bacteria. Microscopic cyanobacteria (picophytoplankton) make up 15% of all the bacteria. Among them, Synechoccus and Prochlorococcus, predominate and constitute the most abundant photosynthetic microbes on earth, contributing more than 50% of the total marine photosynthesis. Of the cultivated bacteria, Roseobacter spp. form about 15% of the total bacteria, while green non sulfur bacteria make up about 6%. C1 Clemson Univ, Dept Biol Sci, Clemson, SC 29634 USA. RP Okafor, N (corresponding author), Clemson Univ, Dept Biol Sci, Clemson, SC 29634 USA. 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TI Biogeographical and Biodiversity Patterns of Marine Planktonic Bacteria Spanning from the South China Sea across the Gulf of Bengal to the Northern Arabian Sea SO MICROBIOLOGY SPECTRUM DT Article AB Understanding the biogeographical and biodiversity patterns of bacterial communities is essential in unraveling their responses to future environmental changes. However, the relationships between marine planktonic bacterial biodiversity and seawater chlorophyll a are largely understudied. Here, we used high-throughput sequencing to study the biodiversity patterns of marine planktonic bacteria across a broad chlorophyll a gradient spanning from the South China Sea across the Gulf of Bengal to the northern Arabian Sea. We found that the biogeographical patterns of marine planktonic bacteria complied with the scenario of homogeneous selection, with chlorophyll a concentration being the key environmental selecting variable of bacteria taxa. The relative abundance of Prochlorococcus, the SAR11 clade, the SAR116 clade, and the SAR86 clade significantly decreased in habitats with high chlorophyll a concentrations (>0.5 mu g/L). Free-living bacteria (FLB) and particle-associated bacteria (PAB) displayed contrasting alpha diversity and chlorophyll a relationships with a positive linear correlation for FLB but a negative correlation for PAB. We further found that PAB had a narrower niche breadth of chlorophyll a than did FLB, with far fewer bacterial taxa being favored at higher chlorophyll a concentrations. Higher chlorophyll a concentrations were linked to the enhanced stochastic drift and reduced beta diversity of PAB but to the weakened homogeneous selection, enhanced dispersal limitation, and increased beta diversity of FLB. Taken together, our findings might broaden our knowledge about the biogeography of marine planktonic bacteria and advance the understanding of bacterial roles in predicting ecosystem functioning under future environmental changes that are derived from eutrophication.IMPORTANCE One of the long-standing interests of biogeography is to explore diversity patterns and uncover their underlying mechanisms. Despite intensive studies on the responses of eukaryotic communities to chlorophyll a concentrations, we know little about how changes in seawater chlorophyll a concentrations affect free-living bacteria (FLB) and particle-associated bacteria (PAB) diversity patterns in natural systems. Our biogeography study demonstrated that marine FLB and PAB displayed contrasting diversity and chlorophyll a relationships and exhibited completely different assembly mechanisms. Our findings broaden our knowledge about the biogeographical and biodiversity patterns of marine planktonic bacteria in nature systems and suggest that PAB and FLB should be considered independently in predicting marine ecosystem functioning under future frequent eutrophication. One of the long-standing interests of biogeography is to explore diversity patterns and uncover their underlying mechanisms. Despite intensive studies on the responses of eukaryotic communities to chlorophyll a concentrations, we know little about how changes in seawater chlorophyll a concentrations affect free-living bacteria (FLB) and particle-associated bacteria (PAB) diversity patterns in natural systems. C1 [Ren, Lijuan; Wu, Chuangfeng; Zhang, Xiufeng; Han, Bo-Ping] Jinan Univ, Dept Ecol, Guangzhou, Peoples R China. [Ren, Lijuan; Wu, Chuangfeng; Zhang, Xiufeng; Han, Bo-Ping] Jinan Univ, Inst Hydrobiol, Guangzhou, Peoples R China. [Ren, Lijuan; Song, Xingyu; Li, Gang; Xia, Xiaomin; Xiang, Chenhui] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou, Peoples R China. [Ren, Lijuan; Song, Xingyu; Li, Gang; Xia, Xiaomin; Xiang, Chenhui] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Sci & Technol Operat Oceanog, Guangzhou, Peoples R China. [Jeppesen, Erik] Univ Chinese Acad Sci, Sino Danish Ctr Educ & Res, Beijing, Peoples R China. [Jeppesen, Erik] Aarhus Univ, Dept Biosci, Silkeborg, Denmark. [Jeppesen, Erik] Middle East Tech Univ, Dept Biol Sci, Limnol Lab, Ankara, Turkiye. [Jeppesen, Erik] Middle East Tech Univ, Ctr Ecosyst Res & Implementat, Ankara, Turkiye. [Wu, Qinglong L.] Southern Marine Sci & Engn Guangdong Lab Guangzho, Ctr Evolut & Conservat Biol, Guangzhou, Peoples R China. [Wu, Qinglong L.] Chinese Acad Sci, Nanjing Inst Geog & Limnol, State Key Lab Lake Sci & Environm, Nanjing, Peoples R China. RP Ren, LJ (corresponding author), Jinan Univ, Dept Ecol, Guangzhou, Peoples R China.; Ren, LJ (corresponding author), Jinan Univ, Inst Hydrobiol, Guangzhou, Peoples R China.; Ren, LJ (corresponding author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou, Peoples R China.; Ren, LJ (corresponding author), Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Sci & Technol Operat Oceanog, Guangzhou, Peoples R China. 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Reintjes, Greta Schattenhofer, Martha Ellrott, Andreas Wulf, Joerg Zeder, Michael Fuchs, Bernhard M. TI Modification of a High-Throughput Automatic Microbial Cell Enumeration System for Shipboard Analyses SO APPLIED AND ENVIRONMENTAL MICROBIOLOGY DT Article AB In the age of ever-increasing "-omics" studies, the accurate and statistically robust determination of microbial cell numbers within often-complex samples remains a key task in microbial ecology. Microscopic quantification is still the only method to enumerate specific subgroups of microbial clades within complex communities by, for example, fluorescence in situ hybridization (FISH). In this study, we improved an existing automatic image acquisition and cell enumeration system and adapted it for usage at high seas on board an oceanographic research ship. The system was evaluated by testing settings such as minimal pixel area and image exposure times ashore under stable laboratory conditions before being brought on board and tested under various wind and wave conditions. The system was robust enough to produce high-quality images even with ship heaves of up to 3m and pitch and roll angles of up to 6.3 degrees. On board the research ship, on average, 25% of the images acquired from plankton samples on filter membranes could be used for cell enumeration. Automated enumeration was highly correlated with manual counts (r(2)>0.9). Even the smallest of microbial cells in the open ocean, members of the alphaproteobacterial SAR11 clade, could be confidently detected and enumerated. The automated image acquisition and cell enumeration system developed here enables an accurate and reproducible determination of microbial cell counts in planktonic samples and allows insight into the abundance and distribution of specific microorganisms already on board within a few hours. IMPORTANCE In this research article, we report on a new system and software pipeline, which allows for an easy and quick image acquisition and the subsequent enumeration of cells in the acquired images. We put this pipeline through vigorous testing and compared it to manual microscopy counts of microbial cells on membrane filters. Furthermore, we tested this system at sea on board a marine research vessel and counted bacteria on board within a few hours after the retrieval of water samples. The imaging and counting system described here has been successfully applied to a number of laboratory-based studies and allowed the quantification of thousands of samples and FISH preparations (see, e.g., H. 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During the multidisciplinary voyage, surface water samples were collected from 19 stations spanning temperate to tropical waters of the EIO (39.5 degrees S to 11.5 degrees S, 110 degrees E). Somewhat surprisingly, a trend of greater DMSPd was measured in ultraoligotrophic (<0.02 mu mol L-1 of nitrate/nitrite), low latitude waters compared to relatively nutrient-rich high latitudes, which contradicts global DMSPd patterns of high concentrations at high latitudes. Additionally, the average DMSPd concentration in EIO samples (17.2 +/- 18.64 nM) was an order of magnitude greater than concentrations previously reported at similar latitudes in the Pacific and Atlantic Oceans, which suggests DMSPd is a readily available food source for microbes in a region that is often considered an ocean desert. The abundances of the bacterial DMSP production gene (dsyB), the DMSP lyase gene (dddP) and phylogenetically diverse DMSP demethylation genes (dmdA subclade A/1, D/all and E/2) were reported for the first time in the EIO region, demonstrating significant shifts in all genes with latitude. The SAR11 dmdA (D/all) gene was the dominant DMSP degradation gene across the transect (3.4 +/- 0.94% of bacteria) and was notably positively correlated to DMSPd, demonstrating a tight coupling between the variables across the 30 degrees transect. Our results also showed greater DMSPd and relative abundance of genes encoding both DMSP degradation pathways (dddP, dmdA A/1 and D/all) within a Leeuwin Current meander when compared to adjacent stations outside of the meander, providing evidence that mesoscale perturbations from the Leeuwin Current can greatly influence the EIO sulfur cycle. 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Coll-Llado, Montserrat Gourdon, Pontus Pascher, Torbjoern Neutze, Richard Pedros-Alio, Carlos Pinhassi, Jarone TI Light stimulates growth of proteorhodopsin-containing marine Flavobacteria SO NATURE DT Article AB Proteorhodopsins are bacterial light-dependent proton pumps. Their discovery within genomic material from uncultivated marine bacterioplankton caused considerable excitement because it indicated a potential phototrophic function within these organisms, which had previously been considered strictly chemotrophic(1). Subsequent studies established that sequences encoding proteorhodopsin are broadly distributed throughout the world's oceans(2-5). Nevertheless, the role of proteorhodopsins in native marine bacteria is still unknown(6). Here we show, from an analysis of the complete genomes of three marine Flavobacteria, that cultivated bacteria in the phylum Bacteroidetes, one of the principal components of marine bacterioplankton, contain proteorhodopsin. Moreover, growth experiments in both natural and artificial seawater ( low in labile organic matter, which is typical of the world's oceans) establish that exposure to light results in a marked increase in the cell yield of one such bacterium (Dokdonia sp. strain MED134) when compared with cells grown in darkness. Thus, our results show that the phototrophy conferred by proteorhodopsin can provide critical amounts of energy, not only for respiration and maintenance but also for active growth of marine bacterioplankton in their natural environment. C1 Univ Kalmar, Dept Biol & Environm Sci, SE-39182 KAlmar, Sweden. Univ La Laguna, Dept Microbiol & Cell Biol, ES-38206 Tenerife, Spain. CSIC, CMIMA, Inst Ciencies Mar, ES-08003 Barcelona, Spain. Chalmers, Dept Biol & Chem Engn, SE-41296 Gothenburg, Sweden. Lund Univ, Kemictr, SE-22100 Lund, Sweden. Univ Gothenburg, Dept Chem Biochem & Biophys, SE-40530 Gothenburg, Sweden. RP Pinhassi, J (corresponding author), Univ Kalmar, Dept Biol & Environm Sci, SE-39182 KAlmar, Sweden. 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Martin, Andrew Maas, Elizabeth W. Ryan, Ken G. TI In situ light responses of the proteorhodopsin-bearing Antarctic sea- ice bacterium, Psychroflexus torques SO ISME JOURNAL DT Article AB Proteorhodopsin (PR) is a wide-spread protein found in many marine prokaryotes. PR allows for the potential conversion of solar energy to ATP, possibly assisting in cellular growth and survival during periods of high environmental stress. PR utilises either blue or green light through a single amino acid substitution. We incubated the PR-bearing bacterium Psychroflexus torquis 50 cm deep within Antarctic sea ice for 13 days, exposing cultures to diurnal fluctuations in light and temperature. Enhanced growth occurred most prominently in cultures incubated under irradiance levels of similar to 50 mu mol photons m(-2) s(-1), suggesting PR provides a strong selective advantage. In addition, cultures grown under blue light yielded over 5.5 times more live cells per photon compared to greenlight incubations. Because P. torquis expresses an apparently 'green-shifted' PR gene variant, this finding infers that the spectral tuning of PR is more complex than previously thought. This study supports the theory that PR provides additional energy to bacteria under sub-optimal conditions, and raises several points of interest to be addressed by future research. C1 [Burr, David J.; Martin, Andrew; Ryan, Ken G.] Victoria Univ Wellington, Sch Biol Sci, POB 600, Wellington 6140, New Zealand. [Burr, David J.; Maas, Elizabeth W.] Natl Inst Water & Atmospher Res NIWA, Wellington, New Zealand. [Martin, Andrew] Univ Tasmania, Inst Marine & Antarctic Sci, Antarctic Gateway Partnership, Hobart, Tas, Australia. [Maas, Elizabeth W.] Minist Primary Ind, Ahuriri, Napier, New Zealand. 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PRs are widely distributed in the oceanic environment and freshwater, but no bacteria with PRs have been isolated from freshwater so far. To facilitate isolation of the bacteria with PR genes, we constructed a vector system that can be used to clone potential PR genes and render color changes when overexpressed in Escherichia coli. Using this method, we successfully isolated a strain with PR gene from freshwater and identified it as Exiguobacterium sp. JL-3. The full length PR gene was then cloned using the SEFA PCR method. Protein sequence alignment showed that JL-3_PR shares high sequence identity (84-89%) with the PRs from Exiguobacterium strains, but low sequence identity (<38%) with other PRs. Surprisingly, we could not detect any proton-pumping activity in the native JL-3 cells and protoplasts, but the recombinant JL-3_PR do pump protons when overexpressed in E.coli. 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Le Huu Cuong Nguyen Dinh Luyen Tran Mai Duc Tran Hong Ha Le Mai Huong Gaerdes, Astrid TI Carrageenophyte-attached and planktonic bacterial communities in two distinct bays of Vietnam: Eutrophication indicators and insights on ice-ice disease SO ECOLOGICAL INDICATORS DT Article AB The composition of the bacterial community of carrageenophyte-attached and planktonic bacteria was studied in two bays of Vietnam with contrasting anthropogenic inputs to assess their role as ecological indicators. Clear differences (similar to 73% dissimilarity) between carrageenophyte-attached bacteria and bacterioplankton were detected in terms of genus composition: mainly Agaribacter, Ruegeria, Alteromonas, the Pir4 lineage and Vibrio for the carrageenophytes and Candidatus Actinomarina, HIMB 11, NS groups and SAR Glades for the bacterioplankton. The copiotrophic nature, potential for complex-polymer degradation, and ability to form and defend biofilms were common features inferred for the carrageenophyte-attached microbiome. Significant differences between the bays were detected in the concentration of most inorganic nutrients. More eutrophic conditions and presumptive wastewater pollution in Cam Ranh (CR) bay were primarily indicated by the dominance of Rubripirellula, Leptobacterium, Hypnocyclicus and Porphyrobacter and their correlations with phosphate. In terms of bacterioplankton, the influence of intensive aquaculture in CR bay was suggested by the dominance of the N55 and N54 marine groups, the SUP05 cluster, Flavobacteriaceae unclassified and SAR 11 Glade III as well as their strong correlations with ammonium and phosphate. The link between silicate and other inorganic nutrients suggests freshwater input in CR bay. Arenicellaceae unclassified and Formosa were also potential indicators of eutrophication. Operational taxonomic units (OTUs) of Marinagarivorans, Cobetia, Vibrio, Alteromonas and Pseudoalteromonas were typical of the carrageenophytes showing ice-ice disease symptoms. Vibrio and Alteromonas were also common among healthy macroalgae, and differences at the OTU level suggested potential succession of species from the healthy to the diseased state. The probable beneficial roles of some bacteria, such as Ruegeria, Cutibacterium and unidentified members of the family Rhizobiaceae, were discussed. This study provides pioneering insights into the bacterial community composition of carrageenophytes and highlights their ecological value as strong indicators of the sources of organic matter, anthropogenic impacts and health status of marine systems. C1 [Kopprio, German A.; Gaerdes, Astrid] Leibniz Ctr Trop Marine Res, Fahrenheitstr 6, D-28359 Bremen, Germany. 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A total of 267 samples were collected from July to December 2016 in the dynamic and shallow southern coastal water of Korea. The variations in BCC mostly depended on planktonic size fraction. Network analysis revealed water depth-dependent co-occurrence patterns of coastal bacterial communities. Higher interspecies connectivity was observed within FL bacteria than NP/MP bacteria, suggesting that FL bacteria with a streamlined genome may need other bacterial metabolites for survival, while the NP/MP copiotrophs may have the self-supporting capacity to produce the vital nutrients. The analysis of topological roles of individual OTUs in the network revealed that several groups of metabolically versatile bacteria (the marine Roseobacters, Flavobacteriales, Desulfobacterales, and SAR406 clade) acted as module hubs in different water depth. 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Bacteria within this layer were sampled using a membrane filter technique and bacterial diversity was compared with that in the underlying pelagic coastal seawater using molecular ecological techniques. 16S rRNA gene libraries of approximate to 500 clones were constructed from both bacterioneuston and the pelagic water samples and representative clones from each library were sequenced for comparison of bacterial diversity. The bacterioneuston was found to have a significantly lower bacterial diversity than the pelagic seawater, with only nine clone types (ecotaxa) as opposed to 46 ecotaxa in the pelagic seawater library. Surprisingly, the bacterioneuston clone library was dominated by 16S rRNA gene sequences affiliated to two groups of organisms, Vibrio spp. which accounted for over 68% of clones and Pseudoalteromonas spp. accounting for 21% of the library. 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Dupont, Christopher L. BE Carlson, CA Giovannoni, SJ TI Microbial Metagenomics: Beyond the Genome SO ANNUAL REVIEW OF MARINE SCIENCE, VOL 3 SE Annual Review of Marine Science DT Review; Book Chapter AB Metagenomics literally means "beyond the genome." Marine microbial metagenomic databases presently comprise similar to 400 billion base pairs of DNA, only similar to 3% of that found in 1 ml of seawater. Very soon a trillion-base-pair sequence run will be feasible, so it is time to reflect on what we have learned from metagenomics. We review the impact of metagenomics on our understanding of marine microbial communities. We consider the studies facilitated by data generated through the Global Ocean Sampling expedition, as well as the revolution wrought at the individual laboratory level through next generation sequencing technologies. We review recent studies and discoveries since 2008, provide a discussion of bioinformatic analyses, including conceptual pipelines and sequence annotation and predict the future of metagenomics, with suggestions of collaborative community studies tailored toward answering some of the fundamental questions in marine microbial ecology. C1 [Gilbert, Jack A.] Plymouth Marine Lab, Plymouth PL1 3DH, Devon, England. [Gilbert, Jack A.] Argonne Natl Lab, Argonne, IL 60439 USA. [Gilbert, Jack A.] Univ Chicago, Dept Ecol & Evolut, Chicago, IL 60637 USA. [Dupont, Christopher L.] J Craig Venter Inst, San Diego, CA 92121 USA. RP Gilbert, JA (corresponding author), Plymouth Marine Lab, Citadel Hill, Plymouth PL1 3DH, Devon, England. 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We revealed novel alternative specificities in MTases that are highly conserved in Alphaproteobacteria, which may enhance our understanding of the co-evolutionary history of the methylation systems and the genomes. Our findings highlight diverse unexplored DNA modifications that potentially affect the ecology and evolution of prokaryotes and viruses in nature. C1 [Hiraoka, Satoshi; Sumida, Tomomi; Nunoura, Takuro] Japan Agcy Marine Earth Sci & Technol JAMSTEC, Res Inst Marine Resources Utilizat, Res Ctr Biosci & Nanosci CeBN, Yokosuka, Kanagawa 2370061, Japan. [Hirai, Miho; Kawagucci, Shinsuke; Yokokawa, Taichi] Japan Agcy Marine Earth Sci & Technol JAMSTEC, Inst Extra Cutting Edge Sci & Technol Avant Garde, Yokosuka, Kanagawa 2370061, Japan. [Toyoda, Atsushi] Natl Inst Genet, Adv Genom Ctr, Mishima, Shizuoka 4118540, Japan. [Kawagucci, Shinsuke] Japan Agcy Marine Earth Sci & Technol JAMSTEC, Res Inst Global Change RIGC, Marine Biodivers & Environm Assessment Res Ctr Bi, Yokosuka, Kanagawa 2370061, Japan. RP Hiraoka, S (corresponding author), Japan Agcy Marine Earth Sci & Technol JAMSTEC, Res Inst Marine Resources Utilizat, Res Ctr Biosci & Nanosci CeBN, Yokosuka, Kanagawa 2370061, Japan. 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Gasol, Josep M. TI Prokaryotic functional gene diversity in the sunlit ocean: Stumbling in the dark SO CURRENT OPINION IN MICROBIOLOGY DT Review AB Prokaryotes are extremely abundant in the ocean where they drive biogeochemical cycles. The recent development and application of -omics techniques has provided an astonishing amount of information revealing the existence of a vast diversity of functional genes and a large heterogeneity within each gene. The big challenge for microbial ecologists is now to understand the ecological relevance of this variability for ecosystem functioning, a question that remains largely understudied. This brief review highlights some of the latest advances in the study of the diversity of biogeochemically relevant functional genes in the sunlit ocean. C1 [Ferrera, Isabel; Sebastian, Marta; Acinas, Silvia G.; Gasol, Josep M.] CSIC, Dept Biol Marina & Oceanog, Inst Ciencies Mar, E-08003 Barcelona, Catalunya, Spain. 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The influence of anthropogenic pressures on plankton communities in well-flushed estuaries has been overlooked. Owing to this ecosystem's short water-residence time, they are foreseen to be mainly composed of microorganisms from neritic-oceanic sources, more resilient to anthropogenic impacts.Aims. This study characterises the bacterioplankton of Urdaibai, a small-drainage estuary located at a UNESCO biosphere reserve, to unravel its annual dynamic and ecological status.Methods. Amplicon sequencing was used to assess the euhaline bacterioplankton shifts and microgAMBI index depicted the ecological status of the estuary.Key results. A differentiated community was identified in upstream waters throughout the year. In addition, organisms commonly associated with faecal waste were detected. The ecological status of the estuary was poor or moderate along the annual cycle, but surprisingly worsened during winter.Conclusions. The above suggests that the bacterial community dynamic in this well-fluxed small-drainage estuary is more complex than anticipated. Importantly, the analysis of pollution-indicative bacteria proved that the water policy of this estuary needs to be further evaluated.Implications. The completion of an efficient sewage treatment system for this protected and touristic estuary is strongly recommended. Incorporating bacterial information to its current monitoring system will constitute a useful and valuable tool. C1 [Aguirre, Mikel; Abad, David; Gutierrez-Muto, Mikel; Langa, Jorge; Estonba, Andone; Zarraonaindia, Iratxe] Univ Basque Country, UPV EHU, Fac Sci & Technol, Dept Genet Phys Anthropol & Anim Physiol, Leioa, Bizkaia, Spain. [Aguirre, Mikel] Anbiolab Anbiotek Biotechnol SL, Derio, Bizkaia, Spain. [Abad, David] Agr Tech Inst Castilla & Leon ITACyL, Mol Biol & Microbiol Lab, Valladolid, Spain. 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Tupper, Ben Sczyrba, Alexander Lauro, Federico M. Martinez-Garcia, Manuel Gonzalez, Jose M. Luo, Haiwei Wright, Jody J. Landry, Zachary C. Hanson, Niels W. Thompson, Brian P. Poulton, Nicole J. Schwientek, Patrick Acinas, Silvia G. Giovannoni, Stephen J. Moran, Mary Ann Hallam, Steven J. Cavicchioli, Ricardo Woyke, Tanja Stepanauskas, Ramunas TI Prevalent genome streamlining and latitudinal divergence of planktonic bacteria in the surface ocean SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA DT Article AB Planktonic bacteria dominate surface ocean biomass and influence global biogeochemical processes, but remain poorly characterized owing to difficulties in cultivation. Using large-scale single cell genomics, we obtained insight into the genome content and biogeography of many bacterial lineages inhabiting the surface ocean. We found that, compared with existing cultures, natural bacterioplankton have smaller genomes, fewer gene duplications, and are depleted in guanine and cytosine, noncoding nucleotides, and genes encoding transcription, signal transduction, and noncytoplasmic proteins. These findings provide strong evidence that genome streamlining and oligotrophy are prevalent features among diverse, free-living bacterioplankton, whereas existing laboratory cultures consist primarily of copiotrophs. The apparent ubiquity of metabolic specialization and mixotrophy, as predicted from single cell genomes, also may contribute to the difficulty in bacterioplankton cultivation. Using metagenome fragment recruitment against single cell genomes, we show that the global distribution of surface ocean bacterioplankton correlates with temperature and latitude and is not limited by dispersal at the time scales required for nucleotide substitution to exceed the current operational definition of bacterial species. Single cell genomes with highly similar small subunit rRNA gene sequences exhibited significant genomic and biogeographic variability, highlighting challenges in the interpretation of individual gene surveys and metagenome assemblies in environmental microbiology. Our study demonstrates the utility of single cell genomics for gaining an improved understanding of the composition and dynamics of natural microbial assemblages. C1 [Swan, Brandon K.; Tupper, Ben; Thompson, Brian P.; Poulton, Nicole J.; Stepanauskas, Ramunas] Bigelow Lab Ocean Sci, East Boothbay, ME 04544 USA. [Sczyrba, Alexander] Univ Bielefeld, Ctr Biotechnol, D-33615 Bielefeld, Germany. [Lauro, Federico M.; Cavicchioli, Ricardo] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia. [Martinez-Garcia, Manuel] Univ Alicante, Dept Physiol Genet & Microbiol, E-03080 Alicante, Spain. [Gonzalez, Jose M.] Univ La Laguna, Dept Microbiol, ES-38206 Tenerife, Spain. 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Despite its importance, the specific influence of viral lysis on seasonal dynamics within aquatic ecosystems remains poorly understood. In this study, we investigated the seasonal dynamics of prokaryotes in the river-estuary-coastal ecosystem surrounding Xiamen Island and explored the environmental factors and virus-mediated cell lysis driving microbial seasonal successions across spatiotemporal scales. A taxon-specific cell lysis was applied to evaluate the contribution of viral lysis to seasonal variations in prokaryotes. Our findings revealed distinct spatiotemporal successions within the prokaryotic community structure, where temporal-related factors, spatial-related factors, and virus-mediated cell lysis contribute comparably to the seasonal variation of prokaryotes. The viral lysis controls on prokaryotic structures were determined by a significant negative correlation between the total microbial community and the cell lysis index (CLI) from amplicon sequence variant (ASV) to order levels. Viral lytic shaping on prokaryotic communities was more pronounced in the estuary-coastal compared to the river region, with similar seasonal variations noted. Specific ASVs, such as ASV3 (Nitrosopumilales), ASV2 (Synechococcales), ASV16 (Nitrosopumilales), and ASV17 (Oceanospirillales) were significantly correlated with CLI, highlighting the pivotal role of viral lysis in their seasonal succession. This study highlights the intricate interplay between microbial populations and viral lysis across spatiotemporal scales, enhancing our understanding of how top-down (virus-mediated cell lysis) and bottom-up (environmental factors) controls drive the seasonal variations in prokaryotic communities. 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Warwick, Phillip E. Zubkov, Mikhail V. TI Low microbial respiration of leucine at ambient oceanic concentration in the mixed layer of the central Atlantic Ocean SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Bacterioplankton are the primary consumers of dissolved organic matter in the ocean, thus the quantification of bacterioplankton production (BP) is essential to our understanding of carbon cycling in the largest ecosystems on Earth. We compared BP, measured as the rate of C-14-leucine or H-3-leucine uptake at close to saturating concentration (20 nmol L-1), with ambient uptake measured from dilution bioassays. We hypothesized that saturation with leucine would lead to its respiration as a carbon source, thereby not truly representing ambient BP. Seawater was collected from the photic zone throughout the Atlantic Ocean. Respiration as a proportion of total consumption (uptake + respiration) of close to ambient (0.4 nmol L-1) and close to saturating (20 nmol L-1) C-14-leucine concentrations were compared. Saturating H-3-leucine additions overestimated ambient leucine uptake at low rates (200% +/- 100% ambient) and underestimated uptake at high rates (90% + 20% ambient). The proportion of total leucine uptake that was respired was threefold higher for 20 nmol L-1 C-14-leucine additions than 0.4 nmol L-1 C-14-leucine additions (15% +/- 8% and 5% +/- 4%, respectively). Consequently, microbial efficiency of leucine assimilation-an indicator of bacterioplankton growth efficiency-was significantly higher and more stable at close to ambient C-14-leucine additions than at saturating concentrations (95% +/- 4% and 85% +/- 8%, respectively). 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TI Prokaryotic responses to a warm temperature anomaly in northeast subarctic Pacific waters SO COMMUNICATIONS BIOLOGY DT Article AB Recent studies on marine heat waves describe water temperature anomalies causing changes in food web structure, bloom dynamics, biodiversity loss, and increased plant and animal mortality. However, little information is available on how water temperature anomalies impact prokaryotes (bacteria and archaea) inhabiting ocean waters. This is a nontrivial omission given their integral roles in driving major biogeochemical fluxes that influence ocean productivity and the climate system. Here we present a time-resolved study on the impact of a large-scale warm water surface anomaly in the northeast subarctic Pacific Ocean, colloquially known as the Blob, on prokaryotic community compositions. Multivariate statistical analyses identified significant depth- and season-dependent trends that were accentuated during the Blob. 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[Ross, Tetjana; Pena, Angelica; Robert, Marie] Inst Ocean Sci Fisheries & Ocean Canada, Sidney, BC, Canada. [McLaughlin, Ryan; Hallam, Steven J.] Univ British Columbia, Grad Program Bioinformat, Vancouver, BC V6T 1Z4, Canada. [Krzywinski, Martin] BC Canc Agcy, Genome Sci Ctr, Vancouver, BC V5Z 4S6, Canada. [Hallam, Steven J.] Univ British Columbia, Genome Sci & Technol Program, 2329 West Mall, Vancouver, BC V6T 1Z4, Canada. [Hallam, Steven J.] Univ British Columbia, Life Sci Inst, Vancouver, BC V6T 1Z3, Canada. [Hallam, Steven J.] Univ British Columbia, ECOSCOPE Training Program, Vancouver, BC V6T 1Z3, Canada. [Traving, Sachia J.] Univ Southern Denmark, HADAL & Nordcee, Dept Biol, Campusvej 55, DK-5230 Odense M, Denmark. [Ho, Grace Y.] Max Planck Inst Marine Microbiol, Celsiusstr 1, D-28359 Bremen, Germany. 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Walsh, David A. TI Linking metagenomics to aquatic microbial ecology and biogeochemical cycles SO LIMNOLOGY AND OCEANOGRAPHY DT Article AB Microbial communities are essential components of aquatic ecosystems through their contribution to food web dynamics and biogeochemical processes. Aquatic microbial diversity is immense and a general challenge is to understand how metabolism and interactions of single organisms shape microbial community dynamics and ecosystem-scale biogeochemical transformations. Metagenomic approaches have developed rapidly, and proven to be powerful in linking microbial community dynamics to biogeochemical processes. In this review, we provide an overview of metagenomic approaches, followed by a discussion on some recent insights they have provided, including those in this special issue. These include the discovery of new taxa and metabolisms in aquatic microbiomes, insights into community assembly and functional ecology as well as evolutionary processes shaping microbial genomes and microbiomes, and the influence of human activities on aquatic microbiomes. Given that metagenomics can now be considered a mature technology where data generation and descriptive analyses are relatively routine and informative, we then discuss metagenomic-enabled research avenues to further link microbial dynamics to biogeochemical processes. These include the integration of metagenomics into well-designed ecological experiments, the use of metagenomics to inform and validate metabolic and biogeochemical models, and the pressing need for ecologically relevant model organisms and simple microbial systems to better interpret the taxonomic and functional information integrated in metagenomes. 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Murdock, Sheryl A. Ji, Qixing Stevens, Catherine J. Grundle, Damian S. Kim Juniper, S. TI Network analysis of 16S rRNA sequences suggests microbial keystone taxa contribute to marine N2O cycling SO COMMUNICATIONS BIOLOGY DT Article AB The mechanisms by which large-scale microbial community function emerges from complex ecological interactions between individual taxa and functional groups remain obscure. We leveraged network analyses of 16S rRNA amplicon sequences obtained over a seven-month timeseries in seasonally anoxic Saanich Inlet (Vancouver Island, Canada) to investigate relationships between microbial community structure and water column N2O cycling. Taxa separately broadly into three discrete subnetworks with contrasting environmental distributions. Oxycline subnetworks were structured around keystone aerobic heterotrophs that correlated with nitrification rates and N2O supersaturations, linking N2O production and accumulation to taxa involved in organic matter remineralization. Keystone taxa implicated in anaerobic carbon, nitrogen, and sulfur cycling in anoxic environments clustered together in a low-oxygen subnetwork that correlated positively with nitrification N2O yields and N2O production from denitrification. Close coupling between N2O producers and consumers in the anoxic basin is indicated by strong correlations between the low-oxygen subnetwork, PICRUSt2-predicted nitrous oxide reductase (nosZ) gene abundances, and N2O undersaturation. This study implicates keystone taxa affiliated with common ODZ groups as a potential control on water column N2O cycling and provides a theoretical basis for further investigations into marine microbial interaction networks. 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We investigated their composition and functional profiles across depths (surface waters 0, 29, and 63 m and bottom waters 100, 150, and 200 m) in the central-eastern Arabian Sea (CEAS) using next-generation sequencing. It was hypothesized that the composition and functional diversity of these communities would be influenced by depth and environmental parameters. Our research showed that microbial communities vary with depth and are shaped by environmental factors like irradiance, temperature, dissolved oxygen, suspended particulate matter, chlorophyll a , and ammonia concentrations. Cyanobacteria ( Prochlorococcus sp) and Mamiellaceae, belonging to picoeukaryotes, exhibited distinct depth- specific distributions up to subsurface chlorophyll maxima (SCM) at 63 m. On the other hand, a community shift in the microbial communities comprising Firmicutes, Bacteroidetes, and Actinobacteria phyla was observed at the deeper water depths. The profiling of functional genes pointed out the expression of carbon fixation by photosynthetic organisms at the surface (0, 29, and 63 m), which shifted to prokaryotic carbon fixation in deeper waters (0, 150, and 200 m). Microcosm experiments (mixing of surface water with water from the SCM) carried out simulating disturbances such as climate change forced mixing (cyclones), revealed shifts in microbial structure and function. It was observed that within 48 h, the carbon fixation activity changed from photosynthetic organisms to prokaryotes and indicated an increase in stress-related biosynthetic pathways such as expression of quorum sensing, biosynthesis of antibiotics, lipopolysaccharides, and secondary metabolites. These findings have implications for predictive modelling of food web dynamics and fisheries management in the context of climate change. 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Donachie, Stuart P. TI Evidence for tropical endemicity in the Deltaproteobacteria Marine Group B/SAR324 bacterioplankton clade SO AQUATIC MICROBIAL ECOLOGY DT Article AB The fine-scale phylogeny of the deeply branching Deltaproteobacteria Marine Group B/SAR324 (MGB/SAR324) bacterioplankton clade was analyzed using partial 16S rRNA gene sequence and internal transcribed spacer (ITS) sequences. Both sets of analyses revealed considerable sequence variation, which, along with bootstrap calculations, strongly support the discrimination of 3 'species' level clusters (i.e. displaying < 98% within-cluster and > 97% between-cluster 16S rRNA gene sequence identity) within this clade, which are here designated as MGB/SAR324 clade groups I and II and the MGB/SAR276 clade. The biogeographical ranges of these 3 clades were analyzed using newly acquired 16S and ITS sequences from Stn ALOHA (22.45 degrees N, 158 degrees W), along with sequences available in the public domain. MGB/SAR324 clade group I and the MBG/SAR276 clade display a clearly restricted distribution, occurring only in tropical and subtropical waters at depths < 250 m. These groups represent the first reported species- and genus-level phylotypes displaying a 'tropical-only' global distribution. MGB/SAR324 clade group II, which has been previously described as ubiquitous, occurs across the latitudinal gradient, but is generally restricted to deep, colder waters in the lower latitudes. Enzyme sequences associated at least with MGB/SAR324 clade group II appear to indicate a role for this group in dissolved organic phosphate cycling. C1 Univ Hawaii, NASA, Astrobiol Inst, Honolulu, HI 96822 USA. Univ Hawaii, Dept Microbiol, Honolulu, HI 96822 USA. RP Brown, MV (corresponding author), Univ Hawaii, NASA, Astrobiol Inst, PSB211,2565 McCarthy Hall, Honolulu, HI 96822 USA. 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The influence of a mesoscale cyclonic eddy on the spatial distribution of bacterioplankton was investigated in a semi-enclosed, oxygen-depleted basin in the Gulf of California, Mexico. Most of the bacterial taxa showed a strong vertical distribution from oxygen-rich surface waters to anoxic bottom waters, but also a horizontal distribution pattern in the upper ocean associated with the eddy presence. Synechococcocales, Flavobacteriales, SAR86, and Actinomarinales were abundant in the euphotic zone within the Gulf of California water mass, whereas SAR324, SAR406, SAR202, SUP05, Arctic97B-4, and Thioglobaceae dominated the bottom layer of this basin within the Subtropical Subsurface water mass. In contrast, bacterial taxa with a preference for mesopelagic waters (Thiomicrospirales, SAR324, SAR202, HOC36, UBA10353 marine group, and Nitrospinales) dominated surface waters of the eddy center where common surface taxa (Synechococcus, SAR86, and Actinomarinales) were scarce. 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Layered distributions were also observed at BATS, although winter deep mixing events disrupted these patterns each year and produced large variations in ecotype abundance. Interestingly, the layered ecotype distributions were regularly reestablished each year after deep mixing subsided at BATS. In addition, Prochlorococcus ecotypes each responded differently to the strong seasonal changes in light, temperature and mixing at BATS, resulting in a reproducible annual succession of ecotype blooms. Patterns of ecotype abundance, in combination with physiological assays of cultured isolates, confirmed that the low-light adapted eNATL could be distinguished from other low-light adapted ecotypes based on its ability to withstand temporary exposure to high-intensity light, a characteristic stress of the surface mixed layer. Finally, total Prochlorococcus and Synechococcus dynamics were compared with similar time series data collected a decade earlier at each location. 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As part of the prokaryotic phototrophic microbial community, both oxygenic phototrophs (OP) and anoxygenic phototrophs (AP) are widely distributed in the ocean and may play a significant role in carbon flow and oxygen production. However, comparative studies of microbial OP and AP have received very little attention, even though their different roles might be important in various marine environments, especially in oxygen minimum zones (OMZ). We explored the spatial distribution of the microbial community in the Baltic Sea, including an OMZ region, with a particular focus on the distribution and activity of OP and AP. We used 16S rRNA amplicon sequencing in combination with a qPCR-based quantification of photosynthesis marker genes. We found that specific bacterial groups dominated surface and intermediate depths, the OMZ, and deep waters, respectively. Salinity, temperature, oxygen, and depth were significant factors explaining the microbial community composition and distribution. 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Gifford, Scott M. Repeta, Daniel J. DeLong, Edward F. TI High molecular weight dissolved organic matter enrichment selects for methylotrophs in dilution to extinction cultures SO ISME JOURNAL DT Article AB The role of bacterioplankton in the cycling of marine dissolved organic matter (DOM) is central to the carbon and energy balance in the ocean, yet there are few model organisms available to investigate the genes, metabolic pathways, and biochemical mechanisms involved in the degradation of this globally important carbon pool. To obtain microbial isolates capable of degrading semi-labile DOM for growth, we conducted dilution to extinction cultivation experiments using seawater enriched with high molecular weight (HMW) DOM. In total, 93 isolates were obtained. Amendments using HMW DOM to increase the dissolved organic carbon concentration 4x (280 mu M) or 10x (700 mu M) the ocean surface water concentrations yielded positive growth in 4-6% of replicate dilutions, whereas <1% scored positive for growth in non-DOM-amended controls. The majority (71%) of isolates displayed a distinct increase in cell yields when grown in increasing concentrations of HMW DOM. Whole-genome sequencing was used to screen the culture collection for purity and to determine the phylogenetic identity of the isolates. Eleven percent of the isolates belonged to the gammaproteobacteria including Alteromonadales (the SAR92 clade) and Vibrio. Surprisingly, 85% of isolates belonged to the methylotrophic OM43 clade of betaproteobacteria, bacteria thought to metabolically specialize in degrading C1 compounds. Growth of these isolates on methanol confirmed their methylotrophic phenotype. Our results indicate that dilution to extinction cultivation enrichede with natural sources of organic substrates has a potential to reveal the previously unsuspected relationships between naturally occurring organic nutrients and the microorganisms that consume them. C1 [Sosa, Oscar A.; DeLong, Edward F.] Univ Hawaii, Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. [Sosa, Oscar A.; Gifford, Scott M.; DeLong, Edward F.] MIT, Dept Civil & Environm Engn, Cambridge, MA 02139 USA. [Sosa, Oscar A.] Woods Hole Oceanog Inst, Dept Biol, Woods Hole, MA 02543 USA. [Repeta, Daniel J.] Woods Hole Oceanog Inst, Dept Marine Chem & Geochem, Woods Hole, MA 02543 USA. RP DeLong, EF (corresponding author), Univ Hawaii, Ctr Microbial Oceanog Res & Educ, Honolulu, HI 96822 USA. 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prosthetic rehabilitation-based gait analysis SO PLOS ONE DT Article AB Quantitative gait analysis is important for understanding the non-typical walking patterns associated with mobility impairments. Conventional linear statistical methods and machine learning (ML) models are commonly used to assess gait performance and related changes in the gait parameters. Nonetheless, explainable machine learning provides an alternative technique for distinguishing the significant and influential gait changes stemming from a given intervention. The goal of this work was to demonstrate the use of explainable ML models in gait analysis for prosthetic rehabilitation in both population- and sample-based interpretability analyses. Models were developed to classify amputee gait with two types of prosthetic knee joints. Sagittal plane gait patterns of 21 individuals with unilateral transfemoral amputations were video-recorded and 19 spatiotemporal and kinematic gait parameters were extracted and included in the models. Four ML models-logistic regression, support vector machine, random forest, and LightGBM-were assessed and tested for accuracy and precision. 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The nitrate maximum was characterized by a high abundance of Thaumarchaeota, the suboxic zone by Gammaproteobacteria Chromatiales, while Epsilonbacteraeota Campylobacterales were abundant at the onset of the sulfidic zone. Compared to the variance in the microbial community, the molecular composition of DOM was relatively uniform across the sampled depths. However, underlying differences in the oxidation state of the DOM molecular formulas showed distinct changes that were linked to the redox zones, possibly connecting autotrophic metabolisms to changes in the DOM composition. In addition, known heterotrophs like Planctomycetes Phycisphaerae and Chloroflexi Anaerolineales were linked to more oxidized molecular forms of DOM, and not to the identified redox zones, suggesting that these fermentative organisms are reliant on newly formed carbon molecules. Our study suggests that the metabolism of autotrophic microbes influences the composition of DOM across the Black Sea water column. 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However, the factors that shape the abundance and potential growth rate of the ecologically distinct prokaryotic subgroups [i.e., high nucleic acid (HNA) and low nucleic acid (LNA) cells] along varying trophic conditions in the ocean remain poorly understood. This study conducted a series of modified dilution experiments to investigate how the abundance and potential growth rate of HNA and LNA prokaryotes and their regulating factors (i.e., protozoan grazing and viral lysis) change along a cross-shore nutrient gradient in the northern South China Sea. The results showed that the abundance of both HNA and LNA cells was significantly positively correlated with the abundance of heterotrophic nanoflagellates and viruses, whereas only HNA abundance exhibited a significant positive correlation with nutrient level. With a decreasing nutrient concentration, the potential growth rate of the HNA subgroup declined significantly, while that of the LNA subgroup was significantly enhanced, leading to an elevated relative potential growth rate of the LNA to HNA subgroup under decreasing nutrient levels. Furthermore, our data revealed different regulatory roles of protozoan grazing and viral lysis on the HNA and LNA subgroups, with HNA suffering higher mortality pressure from grazing than from lysis in contrast to LNA, which experienced equivalent pressures. As the nutrient levels declined, the relative contribution of lysis to the mortality of the HNA subgroup increased significantly, in contrast to the insignificant change in that of the LNA subgroup. Our results indicated the elevated role of LNA cells in the prokaryotic community and the enhanced viral lysis pressure on the total prokaryotes under oligotrophic conditions. 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Although bacterial and archaeal communities at the Juan de Fuca canyon and northward had high similarity, those south of the Juan de Fuca canyon were well differentiated from each other. In contrast, eukaryotic microbial communities exhibited stronger geographic differentiation than bacterial and archaeal communities across the extent of the survey. Seawater parameters that were best predictors of bacterial and archaeal community structure were temperature, pH, and dissolved inorganic nutrients (nitrate, phosphate, silicate), while those that were best predictors of eukaryotic microbial community structure were salinity, dissolved oxygen, total alkalinity, and dissolved inorganic nutrients (nitrite, silicate). Although five bacterial and archaeal indicators for potentially corrosive waters were identified (Colwellia, Nitrosopumilus, Nitrosopelagicus, Sup05 cluster, Sva0996 marine group), no eukaryotic microbial indicators were found. Potentially pathogenic taxa detected in the survey included four disease-causing bacteria for mammals, finfish, and/or shellfish (Coxiella, Flavobacterium, Francisella, Tenacibaculum), sixteen genera of microalgae capable of producing biotoxins, and fifteen parasitic species. This study demonstrates the value of coordinating microbial sampling and analysis with broad-scale oceanographic surveys to generate insights into community structures of these important pelagic trophic levels. C1 [Rhodes, Linda D.; Adams, Nicolaus G.] Natl Ocean & Atmospher Adm NOAA Fisheries, Northwest Fisheries Sci Ctr, Seattle, WA 98115 USA. [Gallego Simon, Ramon] Univ Autonoma Madrid, Biol Dept Univ, Unidad Genet, Madrid, Spain. [Kavanaugh, Maria T.] Oregon State Univ, Coll Earth Ocean & Atmospher Sci, Ocean Ecol & Biogeochem, Corvallis, OR USA. [Alin, Simone R.; Feely, Richard A.] Natl Ocean & Atmospher Adm NOAA, Pacific Marine Environm Lab, Seattle, WA USA. [Rhodes, Linda D.] POB 249, Greenbank, WA USA. 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Bacteriophages play an important role in plankton population dynamics and biogeochemical cycling, but their community dynamics and diversity are still poorly known, especially in fresh waters. 2. We conducted a 1-year investigation of the T4-like bacteriophages in the surface waters of two Western European peri-alpine lakes (lakes Annecy and Bourget), using fingerprinting and cloning-sequencing approaches targeting the major capsid protein g23 gene. 3. Our results suggest that T4-like bacteriophages are diverse (we found several new sequences) and differed substantially between the two lakes. Further, these phages displayed seasonal patterns with marked shifts in community composition. 4. Examining the dynamics of some individual constituents (e. g. 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Schweitzer, Hannah D. Krsmanovic, Stina Campbell, Karley Bernstein, Hans C. TI Diversity and Selection of Surface Marine Microbiomes in the Atlantic-Influenced Arctic SO FRONTIERS IN MICROBIOLOGY DT Article AB Arctic marine environments are experiencing rapid changes due to the polar amplification of global warming. These changes impact the habitat of the cold-adapted microbial communities, which underpin biogeochemical cycles and marine food webs. We comparatively investigated the differences in prokaryotic and microeukaryotic taxa between summer surface water microbiomes sampled along a latitudinal transect from the ice-free southern Barents Sea and into the sea-ice-covered Nansen Basin to disentangle the dominating community (ecological) selection processes driving phylogenetic diversity. The community structure and richness of each site-specific microbiome were assessed in relation to the physical and biogeochemical conditions of the environment. 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Populations in the euphotic zone of this deep (589 m), oligotrophic caldera lake are dominated by two phylogenetic clusters of currently uncultivated bacteria: CL120-10, a newly identified cluster in the verrucomicrobiales, and ACK4 actinomycetes, known as a minor constituent of bacterioplankton in other lakes. Deep-water populations at 300 and 500 m are dominated by a different pair of uncultivated taxa: CL500-11, a novel cluster in the green nonsulfur bacteria, and group I marine crenarchaeota. beta -Proteobacteria, dominant in most other freshwater environments, are relatively rare in Crater Lake (less than or equal to 16% of nonchloroplast bacterial rRNA at all depths). Other taxa identified in Crater Lake libraries include a newly identified candidate bacterial division, ABY1, and a newly identified subcluster, CL0-1, within candidate division OP10. Probe analyses confirmed vertical stratification of several microbial groups, similar to patterns observed in open-ocean systems. 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A total of 4439 clones of 16S rRNA genes from ten 16S rRNA gene libraries were sequenced. More than 97.35% of the sequences represented four bacterial lineages i.e. Alphaproteobacteria, Gammaproteobacteria, Bacteroidetes, and Firmicutes and confirmed their role in scavenging of phytoplankton blooms induced following iron fertilization. The present study demonstrates the response of Firmicutes due to Iron fertilization which was not observed in previous southern ocean Iron fertilization studies. In addition, this study identifies three unique phylogenetic clusters LOHAFEX Cluster 1 (affiliated to Bacteroidetes), 2, and 3 (affiliated to Firmicutes) which were not detected in any of the earlier studies on iron fertilization. The relative abundance of these clusters in response to iron fertilization was different. The increase in abundance of LOHAFEX Cluster 2 and Papillibacter sp. another dominant Firmicutes may imply a role in phytoplankton degradation. 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It has been widely applied in aquatic microbial ecology as a tool to unveil key ecophysiological features, shedding light on relevant ecological issues such as bacterial biomass production, the role of different bacterioplankton groups in the global carbon and sulphur cycle, and, at the same time, providing insights into the life styles and niche differentiation of cosmopolitan members of the aquatic microbial communities. Despite its great potential, its application has remained restricted to a few laboratories around the world, in part due to its reputation as a "difficult technique". Therefore, the objective of this minireview is to highlight the impact of MAR-FISH application on aquatic microbial ecology, and also to provide basic concepts, as well as practical tips, for processing MAR-FISH preparations, thus aiming to contribute to a more widespread application of this powerful method. (c) 2012 Elsevier GmbH. All rights reserved. C1 CURE Univ Republ, Rocha, Uruguay. 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In an ongoing effort to cultivate the uncultured majority of freshwater bacteria, the most abundant freshwater Actinobacteria acI clade has recently been cultivated from Lake Soyang through catalase-supplemented high-throughput cultivation based on dilution-to-extinction. This method involves physical isolation of target microbes from mixed populations, culture media simulating natural habitats, and removal of toxic compounds. In this protocol, we describe detailed procedures for isolating freshwater oligotrophic microbes, as well as the essence of the dilution-to-extinction culturing. As a case study employing the catalase-supplemented dilution-to-extinction protocol, we also report a cultivation trial using a water sample collected from Lake Soyang. Of the 480 cultivation wells inoculated with a single lake-water sample, 75 new acI strains belonging to 8 acI tribes (acI-A1, A2, A4, A5, A6, A7, B1, B4, C1, and C2) were cultivated, and each representative strain per subclade could be revived from glycerol stocks. These cultivation results demonstrate that the protocol described in this study is efficient in isolating freshwater bacterioplankton harboring streamlined genomes. C1 [Kim, Suhyun; Park, Miri S.; Song, Jaeho; Kang, Ilnam; Cho, Jang-Cheon] Inha Univ, Dept Biol Sci, Incheon 22212, South Korea. RP Cho, JC (corresponding author), Inha Univ, Dept Biol Sci, Incheon 22212, South Korea. 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Viruses are also extraordinarily diverse, which is evident in the types of relationships they establish with their host, their strategies to store and replicate their genetic information and the enormous diversity of genes they contain. A viral population, especially if it corresponds to a virus with an RNA genome, can contain an array of sequence variants that greatly exceeds what is present in most cell populations. The fact that viruses always need cellular resources to multiply means that they establish very close interactions with cells. Although in the short term these relationships may appear to be negative for life, it is evident that they can be beneficial in the long term. Viruses are one of the most powerful selective pressures that exist, accelerating the evolution of defense mechanisms in the cellular world. They can also exchange genetic material with the host during the infection process, providing organisms with capacities that favor the colonization of new ecological niches or confer an advantage over competitors, just to cite a few examples. In addition, viruses have a relevant participation in the biogeochemical cycles of our planet, contributing to the recycling of the matter necessary for the maintenance of life. Therefore, although viruses have traditionally been excluded from the tree of life, the structure of this tree is largely the result of the interactions that have been established throughout the intertwined history of the cellular and the viral worlds. We do not know how other possible biospheres outside our planet could be, but it is clear that viruses play an essential role in the terrestrial one. Therefore, they must be taken into account both to improve our understanding of life that we know, and to understand other possible lives that might exist in the cosmos. 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Here, we assessed resource partitioning among freshwater bacterial groups from two oligotrophic lakes using four types of organic substrates as compound models. Substrate uptake patterns were examined by microautoradiography combined with catalysed reporter deposition fluorescent in situ hybridization. Four large taxonomic groups were found in the lakes, but Actinobacteria (AcI lineage) and Betaproteobacteria (R-BTcluster) dominated the bacterial assemblage. Monomers containing nitrogen and/or phosphorus were preferred over the ones containing only carbon. All groups were able to incorporate amino acids, adenosine triphosphate and glucose. However, acetate was only taken up by approximate to 10-12% of bacteria, and its uptake was not detected in Cytophaga-Flavobacteria. Apart from acetate, the contribution of a particular bacterial group to the uptake of a substrate was proportional to its relative abundance. 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The combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR) is currently the most widely applied tool for revealing physiological properties of microorganisms in their natural environment with single-cell resolution. For example, this technique has been used in wastewater treatment and marine systems to describe the functional properties of newly discovered species, and to identify microorganisms responsible for key physiological processes. Recently, the scope of FISH-MAR was extended by rendering it quantitative and by combining it with microelectrode measurements or stable isotope probing. Isotope arrays have also been developed that exploit the parallel detection offered by DNA microarrays to measure incorporation of labelled substrate into the rRNA of many community members in a single experiment. C1 Univ Vienna, Dept Microbial Ecol, A-1090 Vienna, Austria. Aalborg Univ, Dept Life Sci, DK-9000 Aalborg, Denmark. 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Gasol, Josep M. Pinhassi, Jarone TI Structuring of bacterioplankton communities by specific dissolved organic carbon compounds SO ENVIRONMENTAL MICROBIOLOGY DT Article AB The main role of microorganisms in the cycling of the bulk dissolved organic carbon pool in the ocean is well established. Nevertheless, it remains unclear if particular bacteria preferentially utilize specific carbon compounds and whether such compounds have the potential to shape bacterial community composition. Enrichment experiments in the Mediterranean Sea, Baltic Sea and the North Sea (Skagerrak) showed that different low-molecular-weight organic compounds, with a proven importance for the growth of marine bacteria (e.g. amino acids, glucose, dimethylsulphoniopropionate, acetate or pyruvate), in most cases differentially stimulated bacterial growth. Denaturing gradient gel electrophoresis fingerprints and 16S rRNA gene sequencing revealed that some bacterial phylotypes that became abundant were highly specific to enrichment with specific carbon compounds (e.g. Acinetobacter sp. B1-A3 with acetate or Psychromonas sp. B3-U1 with glucose). In contrast, other phylotypes increased in relative abundance in response to enrichment with several, or all, of the investigated carbon compounds (e.g. Neptuniibacter sp. M2-A4 with acetate, pyruvate and dimethylsulphoniopropionate, and Thalassobacter sp. M3-A3 with pyruvate and amino acids). Furthermore, different carbon compounds triggered the development of unique combinations of dominant phylotypes in several of the experiments. These results suggest that bacteria differ substantially in their abilities to utilize specific carbon compounds, with some bacteria being specialists and others having a more generalist strategy. Thus, changes in the supply or composition of the dissolved organic carbon pool can act as selective forces structuring bacterioplankton communities. C1 [Gomez-Consarnau, Laura; Lindh, Markus V.; Pinhassi, Jarone] Linnaeus Univ, Sch Nat Sci, Kalmar, Sweden. [Gasol, Josep M.] CSIC, Inst Ciencies Mar, Barcelona, Catalonia, Spain. RP Pinhassi, J (corresponding author), Linnaeus Univ, Sch Nat Sci, Kalmar, Sweden. 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Dithugoe, Choaro D. Bezuidt, Oliver K. Makhalanyane, Thulani P. TI Microbial ecology of the Southern Ocean SO FEMS MICROBIOLOGY ECOLOGY DT Review AB The Southern Ocean (SO) distributes climate signals and nutrients worldwide, playing a pivotal role in global carbon sequestration. Microbial communities are essential mediators of primary productivity and carbon sequestration, yet we lack a comprehensive understanding of microbial diversity and functionality in the SO. Here, we examine contemporary studies in this unique polar system, focusing on prokaryotic communities and their relationships with other trophic levels (i.e. phytoplankton and viruses). Strong seasonal variations and the characteristic features of this ocean are directly linked to community composition and ecosystem functions. Specifically, we discuss characteristics of SO microbial communities and emphasise differences from the Arctic Ocean microbiome. We highlight the importance of abundant bacteria in recycling photosynthetically derived organic matter. These heterotrophs appear to control carbon flux to higher trophic levels when light and iron availability favour primary production in spring and summer. Conversely, during winter, evidence suggests that chemolithoautotrophs contribute to prokaryotic production in Antarctic waters. We conclude by reviewing the effects of climate change on marine microbiota in the SO. C1 [Castillo, Diego J.; Dithugoe, Choaro D.; Bezuidt, Oliver K.; Makhalanyane, Thulani P.] Univ Pretoria, Dept Biochem Genet & Microbiol, Microbiome Res Grp, ZA-0028 Pretoria, South Africa. [Castillo, Diego J.; Dithugoe, Choaro D.; Bezuidt, Oliver K.; Makhalanyane, Thulani P.] Univ Pretoria, Dept Sci, ZA-0028 Pretoria, South Africa. [Castillo, Diego J.; Dithugoe, Choaro D.; Bezuidt, Oliver K.; Makhalanyane, Thulani P.] Univ Pretoria, Innovat South African Res Chair Marine Microbiom, Dept Biochem Genet & Microbiol, ZA-0028 Pretoria, South Africa. RP Makhalanyane, TP (corresponding author), Univ Pretoria, Dept Biochem Genet & Microbiol, Microbiome Res Grp, ZA-0028 Pretoria, South Africa.; Makhalanyane, TP (corresponding author), Univ Pretoria, Dept Sci, ZA-0028 Pretoria, South Africa.; Makhalanyane, TP (corresponding author), Univ Pretoria, Innovat South African Res Chair Marine Microbiom, Dept Biochem Genet & Microbiol, ZA-0028 Pretoria, South Africa. 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By high-throughput metagenomics, we were able to reconstruct the genomes of representative microbes. The gross community structure (at the phylum level) was different from the structure of typical marine and freshwater communities in temperate open oceans, with the Caspian Sea having freshwater-like amounts of Actinobacteria and Alphaproteobacteria, while Gammaproteobacteria and Betaproteobacteria were present at intermediate levels. We assembled the genomes of several groups and provide detailed descriptions of partial genomes from Actinobacteria, Thaumarchaea, and Alphaproteobacteria. Most belonged to hitherto unknown groups, although they were related to either marine or freshwater groups. The phylogenetic placement of the Caspian genomes indicates that the organisms have multiple and separate phylogenetic origins and that they are related to organisms with both freshwater and marine lineages. Comparative recruitment from global aquatic metagenomes indicated that most Caspian microbes are endemic. However, some Caspian genomes were recruited significantly from either marine water (a member of the Alphaproteobacteria) or freshwater (a member of the Actinobacteria). Reciprocally, some genomes of other origins, such as the marine thaumarchaeon "Candidatus Nitrosopelagicus" or the actinobacterium "Candidatus Actinomarina," were recruited from the Caspian Sea, indicating some degree of overlap with the microbiota of other water bodies. Some of these microbes seem to have a remarkably widespread geographic and environmental distribution. C1 [Mehrshad, Maliheh; Amoozegar, Mohammad Ali] Univ Tehran, Fac Biol, Dept Microbiol, Extremophiles Lab, Tehran, Iran. [Mehrshad, Maliheh; Amoozegar, Mohammad Ali] Univ Tehran, Ctr Excellence Phylogeny Living Organisms, Coll Sci, Tehran, Iran. [Ghai, Rohit; Rodriguez-Valera, Francisco] Univ Miguel Hernandez, Evolutionary Genom Grp, Alacant, Spain. [Fazeli, Seyed Abolhassan Shahzadeh] ACECR, Iranian Biol Resource Ctr IBRC, Microorganisms Bank, Tehran, Iran. [Fazeli, Seyed Abolhassan Shahzadeh] Univ Sci & Culture, Fac Basic Sci & Adv Technol Biol, Dept Mol & Cellular Biol, Tehran, Iran. [Ghai, Rohit] Acad Sci Czech Republic, Inst Hydrobiol, Ctr Biol, Vvi, Ceske Budejovice, Czech Republic. RP Amoozegar, MA (corresponding author), Univ Tehran, Fac Biol, Dept Microbiol, Extremophiles Lab, Tehran, Iran.; Amoozegar, MA (corresponding author), Univ Tehran, Ctr Excellence Phylogeny Living Organisms, Coll Sci, Tehran, Iran. 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Le Romancer, Marc Wauters, Nina Jacquet, Stephan TI Deciphering the virus-to-prokaryote ratio (VPR): insights into virus-host relationships in a variety of ecosystems SO BIOLOGICAL REVIEWS DT Review AB The discovery of the numerical importance of viruses in a variety of (aquatic) ecosystems has changed our perception of their importance in microbial processes. Bacteria and Archaea undoubtedly represent the most abundant cellular life forms on Earth and past estimates of viral numbers (represented mainly by viruses infecting prokaryotes) have indicated abundances at least one order of magnitude higher than that of their cellular hosts. Such dominance has been reflected most often by the virus-to-prokaryote ratio (VPR), proposed as a proxy for the relationship between viral and prokaryotic communities. VPR values have been discussed in the literature to express viral numerical dominance (or absence of it) over their cellular hosts, but the ecological meaning and interpretation of this ratio has remained somewhat nebulous or contradictory. We gathered data from 210 publications (and additional unpublished data) on viral ecology with the aim of exploring VPR. The results are presented in three parts: the first consists of an overview of the minimal, maximal and calculated average VPR values in an extensive variety of different environments. Results indicate that VPR values fluctuate over six orders of magnitude, with variations observed within each ecosystem. The second part investigates the relationship between VPR and other indices, in order to assess whether VPR can provide insights into virus-host relationships. A positive relationship was found between VPR and viral abundance (VA), frequency of visibly infected cells (FVIC), burst size (BS), frequency of lysogenic cells (FLC) and chlorophyll a (Chl a) concentration. An inverse relationship was detected between VPR and prokaryotic abundance (PA) (in sediments), prokaryotic production (PP) and virus-host contact rates (VCR) as well as salinity and temperature. No significant relationship was found between VPR and viral production (VP), fraction of mortality from viral lysis (FMVL), viral decay rate (VDR), viral turnover (VT) or depth. Finally, we summarize our results by proposing two scenarios in two contrasting environments, based on current theories on viral ecology as well as the present results. We conclude that since VPR fluctuates in every habitat for different reasons, as it is linked to a multitude of factors related to virus-host dynamics, extreme caution should be used when inferring relationships between viruses and their hosts. Furthermore, we posit that the VPR is only useful in specific, controlled conditions, e. g. for the monitoring of fluctuations in viral and host abundance over time. C1 [Parikka, Kaarle J.; Le Romancer, Marc] Inst Univ Europeen Mer, Lab Microbiol Extreme Environm, F-29280 Plouzane, France. [Parikka, Kaarle J.] Queen Astrid Mil Hosp, Belgian Dept Def, LabMCT, B-1120 Brussels, Belgium. [Wauters, Nina] Univ Libre Bruxelles, Biol Evolut & Ecol, B-1050 Brussels, Belgium. [Jacquet, Stephan] INRA CARRTEL, F-74200 Thonon Les Bains, France. RP Jacquet, S (corresponding author), INRA CARRTEL, F-74200 Thonon Les Bains, France. 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