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  • 1
    Electronic Resource
    Electronic Resource
    Bacterial diversity in a subseafloor habitat following a deep-sea volcanic eruption (2003)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 43 (2003), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The bacterial diversity in a diffuse flow hydrothermal vent habitat at Axial Volcano, Juan de Fuca Ridge was examined shortly after an eruptive event in 1998 and again in 1999 and 2000 using PCR-amplified 16S rRNA gene sequence analyses. While considerable overlap with deep-sea background seawater was found within the α- and γ-proteobacteria, unique subseafloor phylotypes were distinguishable. These included diverse members of the ε-proteobacteria, high temperature groups such as Desulfurobacterium, Gram-positive bacteria, and members of novel candidate divisions WS6 and ABY1. Phylotype richness was highest in the particle-attached populations from all three sampling periods, and diversity appeared to increase over that time, particularly among the ε-proteobacteria. A preliminary model of the subseafloor is presented that relates microbial diversity to temperature, chemical characteristics of diffuse flow fluids and the degree of mixing with seawater.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.2003.tb01080.x
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  • 2
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    Accuracy and quality of massively parallel DNA pyrosequencing (2007)
    BioMed Central
    Details
    Publication Date: 2022-05-25
    Description: © 2007 Huse et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The definitive version was published in Genome Biology 8 (2007): R143, doi:10.1186/gb-2007-8-7-r143.
    Description: Additional data file 1 is a fasta file of the 43 known sequences used. Additional data file 2 is a gzip-compressed fasta file of the sequences output by the GS20. These sequences correspond to those included in Additional data files 3, 4, 5 but include only the final sequence information. Additional data files 3, 4, 5 are three compressed text files representing the text translations of the original GS20 binary output (sff) files for all of the sequencing used in the analysis, including sequence, flowgram and other run information. GS20 data are reported by region of the PicoTiterPlate™; we sequenced three plate regions.
    Description: Massively parallel pyrosequencing systems have increased the efficiency of DNA sequencing, although the published per-base accuracy of a Roche GS20 is only 96%. In genome projects, highly redundant consensus assemblies can compensate for sequencing errors. In contrast, studies of microbial diversity that catalogue differences between PCR amplicons of ribosomal RNA genes (rDNA) or other conserved gene families cannot take advantage of consensus assemblies to detect and minimize incorrect base calls. We performed an empirical study of the per-base error rate for the Roche GS20 system using sequences of the V6 hypervariable region from cloned microbial ribosomal DNA (tag sequencing). We calculated a 99.5% accuracy rate in unassembled sequences, and identified several factors that can be used to remove a small percentage of low-quality reads, improving the accuracy to 99.75% or better. By using objective criteria to eliminate low quality data, the quality of individual GS20 sequence reads in molecular ecological applications can surpass the accuracy of traditional capillary methods.
    Description: This work was supported by National Aeronautics and Space Administration Astrobiology Institute Cooperative Agreement NNA04CC04A (to MLS), subcontracts from the Woods Hole Center for Oceans and Human Health from the National Institutes of Health and National Science Foundation (NIH/NIEHS 1 P50 ES012742-01 and NSF/OCE 0430724-J Stegeman PI to HGM and MLS), grants from the WM Keck Foundation and the G Unger Vetlesen Foundation (to MLS), and a National Research Council Research Associateship Award (to JAH).
    Repository Name: Woods Hole Open Access Server
    Type: Article
    Format: application/pdf
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  • 3
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    Effect of PCR amplicon size on assessments of clone library microbial diversity and community structure (2009)
    Details
    Publication Date: 2022-05-25
    Description: Author Posting. © The Authors, 2009. This is the author's version of the work. It is posted here by permission of Society for Applied Microbiology and Blackwell Publishing for personal use, not for redistribution. The definitive version was published in Environmental Microbiology 11 (2009): 1292-1302, doi:10.1111/j.1462-2920.2008.01857.x.
    Description: PCR-based surveys of microbial communities commonly use regions of the small subunit ribosomal RNA (SSU rRNA) gene to determine taxonomic membership and estimate total diversity. Here we show that the length of the target amplicon has a significant effect on assessments of microbial richness and community membership. Using OTU- and taxonomy-based tools, we compared the V6 hypervariable region of the bacterial SSU rRNA gene of three amplicon libraries of ca. 100 base pair (bp), 400bp, and 1000bp from each of two hydrothermal vent fluid samples. We found that the smallest amplicon libraries contained more unique sequences, higher diversity estimates, and a different community structure than the other two libraries from each sample. We hypothesize that a combination of polymerase dissociation, cloning bias, and mis-priming due to secondary structure accounts for the differences. While this relationship is not linear, it is clear that the smallest amplicon libraries contained more different types of sequences, and accordingly, more diverse members of the community. Because divergent and lower abundant taxa can be more readily detected with smaller amplicons, they may provide better assessments of total community diversity and taxonomic membership than longer amplicons in molecular studies of microbial communities.
    Description: This work was supported by a National Research Council Research Associateship Award and L'Oréal USA Fellowship (J.A.H.), NASA Astrobiology Institute Cooperative Agreement NNA04CC04A (M.L.S.), the Alfred P. Sloan Foundation's ICoMM field project, and a subcontract from the Woods Hole Center for Oceans and Human Health from the National Institutes of Health and the National Science Foundation (NIH/NIEHS 1 P50 ES012742-01 and NSF/OCE 0430724; J.Stegeman, PI to HGM and MLS).
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 4
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    Seasonal succession and spatial patterns of Synechococcus microdiversity in a salt marsh estuary revealed through 16S rRNA gene oligotyping (2017)
    Frontiers Media
    Details
    Publication Date: 2022-05-25
    Description: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Frontiers in Microbiology 8 (2017): 1496, doi:10.3389/fmicb.2017.01496.
    Description: Synechococcus are ubiquitous and cosmopolitan cyanobacteria that play important roles in global productivity and biogeochemical cycles. This study investigated the fine scale microdiversity, seasonal patterns, and spatial distributions of Synechococcus in estuarine waters of Little Sippewissett salt marsh (LSM) on Cape Cod, MA. The proportion of Synechococcus reads was higher in the summer than winter, and higher in coastal waters than within the estuary. Variations in the V4–V6 region of the bacterial 16S rRNA gene revealed 12 unique Synechococcus oligotypes. Two distinct communities emerged in early and late summer, each comprising a different set of statistically co-occurring Synechococcus oligotypes from different clades. The early summer community included clades I and IV, which correlated with lower temperature and higher dissolved oxygen levels. The late summer community included clades CB5, I, IV, and VI, which correlated with higher temperatures and higher salinity levels. Four rare oligotypes occurred in the late summer community, and their relative abundances more strongly correlated with high salinity than did other co-occurring oligotypes. The analysis revealed that multiple, closely related oligotypes comprised certain abundant clades (e.g., clade 1 in the early summer and clade CB5 in the late summer), but the correlations between these oligotypes varied from pair to pair, suggesting they had slightly different niches despite being closely related at the clade level. Lack of tidal water exchange between sampling stations gave rise to a unique oligotype not abundant at other locations in the estuary, suggesting physical isolation plays a role in generating additional microdiversity within the community. Together, these results contribute to our understanding of the environmental and ecological factors that influence patterns of Synechococcus microbial community composition over space and time in salt marsh estuarine waters.
    Description: This work was supported through a subcontract from the Woods Hole Center for Oceans and Human Health, from the National Institutes of Health and the National Science Foundation (NIH/NIEHS 1 P50 ES012742-01 and NSF/OCE 0430724), a National Research Council Research Associateship Award and L'Oreal USA Fellowship (JH), an Alfred P. Sloan Research Fellowship in Ocean Sciences and the Clare Boothe Luce Program (KM), NASA Astrobiology Institute Cooperative Agreement NNA04CC04A (MS), the Alfred P. Sloan Foundation's ICoMM field project, and the W. M. Keck Foundation.
    Repository Name: Woods Hole Open Access Server
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  • 5
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    Salt marsh sediment bacterial communities maintain original population structure after transplantation across a latitudinal gradient (2018)
    PeerJ
    Details
    Publication Date: 2022-05-25
    Description: © The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PeerJ 6 (2018): e4735, doi:10.7717/peerj.4735.
    Description: Dispersal and environmental selection are two of the most important factors that govern the distributions of microbial communities in nature. While dispersal rates are often inferred by measuring the degree to which community similarity diminishes with increasing geographic distance, determining the extent to which environmental selection impacts the distribution of microbes is more complex. To address this knowledge gap, we performed a large reciprocal transplant experiment to simulate the dispersal of US East Coast salt marsh Spartina alterniflora rhizome-associated microbial sediment communities across a latitudinal gradient and determined if any shifts in microbial community composition occurred as a result of the transplantation. Using bacterial 16S rRNA gene sequencing, we did not observe large-scale changes in community composition over a five-month S. alterniflora summer growing season and found that transplanted communities more closely resembled their origin sites than their destination sites. Furthermore, transplanted communities grouped predominantly by region, with two sites from the north and three sites to the south hosting distinct bacterial taxa, suggesting that sediment communities transplanted from north to south tended to retain their northern microbial distributions, and south to north maintained a southern distribution. A small number of potential indicator 16S rRNA gene sequences had distributions that were strongly correlated to both temperature and nitrogen, indicating that some organisms are more sensitive to environmental factors than others. These results provide new insight into the microbial biogeography of salt marsh sediments and suggest that established bacterial communities in frequently-inundated environments may be both highly resistant to invasion and resilient to some environmental shifts. However, the extent to which environmental selection impacts these communities is taxon specific and variable, highlighting the complex interplay between dispersal and environmental selection for microbial communities in nature.
    Description: This research was conducted in the National Estuarine Research Reserve System under an award from the Estuarine Reserves Division, Office of Ocean and Coastal Resource Management, National Ocean Service, and National Oceanic and Atmospheric Administration. Support was also provided through funding to Julie Huber from a Brown-MBL Partnership SEED award, the Neal Cornell Endowed Research Fund, and the NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564). Additional funding was provided to Sarah Corman-Crosby by the National Park Service George Melendez Wright Climate Change Fellowship.
    Keywords: Microbial ecosystems ; Dispersal ; Bacteria ; Salt marsh ; Biogeography ; Reciprocal transplant
    Repository Name: Woods Hole Open Access Server
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  • 6
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    Exploring microbial diversity and taxonomy using SSU rRNA hypervariable tag sequencing (2009)
    Public Library of Science
    Details
    Publication Date: 2022-05-25
    Description: © 2008 Huse et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS Genetics 4 (2008): e1000255, doi:10.1371/journal.pgen.1000255.
    Description: Massively parallel pyrosequencing of hypervariable regions from small subunit ribosomal RNA (SSU rRNA) genes can sample a microbial community two or three orders of magnitude more deeply per dollar and per hour than capillary sequencing of full-length SSU rRNA. As with full-length rRNA surveys, each sequence read is a tag surrogate for a single microbe. However, rather than assigning taxonomy by creating gene trees de novo that include all experimental sequences and certain reference taxa, we compare the hypervariable region tags to an extensive database of rRNA sequences and assign taxonomy based on the best match in a Global Alignment for Sequence Taxonomy (GAST) process. The resulting taxonomic census provides information on both composition and diversity of the microbial community. To determine the effectiveness of using only hypervariable region tags for assessing microbial community membership, we compared the taxonomy assigned to the V3 and V6 hypervariable regions with the taxonomy assigned to full-length SSU rRNA sequences isolated from both the human gut and a deep-sea hydrothermal vent. The hypervariable region tags and full-length rRNA sequences provided equivalent taxonomy and measures of relative abundance of microbial communities, even for tags up to 15% divergent from their nearest reference match. The greater sampling depth per dollar afforded by massively parallel pyrosequencing reveals many more members of the “rare biosphere” than does capillary sequencing of the full-length gene. In addition, tag sequencing eliminates cloning bias and the sequences are short enough to be completely sequenced in a single read, maximizing the number of organisms sampled in a run while minimizing chimera formation. This technique allows the cost-effective exploration of changes in microbial community structure, including the rare biosphere, over space and time and can be applied immediately to initiatives, such as the Human Microbiome Project.
    Description: Woods Hole Center for Oceans and Human Health from the National Institutes of Health and National Science Foundation (NIH/NIEHS 1 P50 ES012742-01 and NSF/OCE 0430724-J Stegeman PI to MLS). NIH Director's Pioneer Award and Doris Duke Distinguished Clinical Scientist Award to DAR.
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  • 7
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    Trophic regions of a hydrothermal plume dispersing away from an ultramafic-hosted vent-system : Von Damm vent-site, Mid-Cayman Rise (2013)
    John Wiley & Sons
    Details
    Publication Date: 2022-05-25
    Description: Author Posting. © American Geophysical Union, 2013. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geochemistry, Geophysics, Geosystems 14 (2013): 317–327, doi:10.1002/ggge.20063.
    Description: Deep-sea ultramafic-hosted vent systems have the potential to provide large amounts of metabolic energy to both autotrophic and heterotrophic microorganisms in their dispersing hydrothermal plumes. Such vent-systems release large quantities of hydrogen and methane to the water column, both of which can be exploited by autotrophic microorganisms. Carbon cycling in these hydrothermal plumes may, therefore, have an important influence on open-ocean biogeochemistry. In this study, we investigated an ultramafic-hosted system on the Mid-Cayman Rise, emitting metal-poor and hydrogen sulfide-, methane-, and hydrogen-rich hydrothermal fluids. Total organic carbon concentrations in the plume ranged between 42.1 and 51.1 μM (background = 43.2 ± 0.7 μM (n = 5)) and near-field plume samples with elevated methane concentrations imply the presence of chemoautotrophic primary production and in particular methanotrophy. In parts of the plume characterized by persistent potential temperature anomalies but lacking elevated methane concentrations, we found elevated organic carbon concentrations of up to 51.1 μM, most likely resulting from the presence of heterotrophic communities, their extracellular products and vent larvae. Elevated carbon concentrations up to 47.4 μM were detected even in far-field plume samples. Within the Von Damm hydrothermal plume, we have used our data to hypothesize a microbial food web in which chemoautotrophy supports a heterotrophic community of microorganisms. Such an active microbial food web would provide a source of labile organic carbon to the deep ocean that should be considered in any future studies evaluating sources and sinks of carbon from hydrothermal venting to the deep ocean.
    Description: The research reported in this paper was supported by ship time and support provided by NOAA’s Office of Ocean Exploration and Research and the Office of Marine and Aviation Operations and NSF’s Division of Ocean Sciences (Grant OCE-1061863) and by further shore-based research from both the National Science Foundation (NSF OCE-1061863) and NASA’s ASTEP Program (Grant # NNX09AB75G). The contributions of SB and MC were carried out at the Jet Propulsion Laboratory (JPL), California Institute of Technology, under contract with the National Aeronautics and Space Administration (NASA), with support from the NASA ASTEP Program.
    Description: 2013-08-22
    Keywords: Hydrothermal ; Food web ; Microorganisms ; Plume ; Carbon ; Ultramafic
    Repository Name: Woods Hole Open Access Server
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  • 8
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    Coupled RNA-SIP and metatranscriptomics of active chemolithoautotrophic communities at a deep-sea hydrothermal vent (2016)
    Nature Publishing Group
    Details
    Publication Date: 2022-05-25
    Description: © The International Society for Microbial Ecology, 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in ISME Journal 10 (2016): 1925–1938, doi:10.1038/ismej.2015.258.
    Description: The chemolithoautotrophic microbial community of the rocky subseafloor potentially provides a large amount of organic carbon to the deep ocean, yet our understanding of the activity and metabolic complexity of subseafloor organisms remains poorly described. A combination of metagenomic, metatranscriptomic, and RNA stable isotope probing (RNA-SIP) analyses were used to identify the metabolic potential, expression patterns, and active autotrophic bacteria and archaea and their pathways present in low-temperature hydrothermal fluids from Axial Seamount, an active submarine volcano. Metagenomic and metatranscriptomic results showed the presence of genes and transcripts for sulfur, hydrogen, and ammonium oxidation, oxygen respiration, denitrification, and methanogenesis, as well as multiple carbon fixation pathways. In RNA-SIP experiments across a range of temperatures under reducing conditions, the enriched 13C fractions showed differences in taxonomic and functional diversity. At 30 °C and 55 °C, Epsilonproteobacteria were dominant, oxidizing hydrogen and primarily reducing nitrate. Methanogenic archaea were also present at 55 °C, and were the only autotrophs present at 80 °C. Correspondingly, the predominant CO2 fixation pathways changed from the reductive tricarboxylic acid (rTCA) cycle to the reductive acetyl-CoA pathway with increasing temperature. By coupling RNA-SIP with meta-omics, this study demonstrates the presence and activity of distinct chemolithoautotrophic communities across a thermal gradient of a deep-sea hydrothermal vent.
    Description: This work was funded by the Gordon and Betty Moore Foundation Grant GBMF3297 and NSF Center for Dark Energy Biosphere Investigations (C-DEBI) (OCE-0939564). The data collected in this study is based upon work supported by the Schmidt Ocean Institute during cruise FK010-2013 aboard R/V Falkor.
    Repository Name: Woods Hole Open Access Server
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  • 9
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    Genomic variation in microbial populations inhabiting the marine subseafloor at deep-sea hydrothermal vents (2017)
    Nature Publishing Group
    Details
    Publication Date: 2022-05-25
    Description: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Nature Communications 8 (2017): 1114, doi:10.1038/s41467-017-01228-6.
    Description: Little is known about evolutionary drivers of microbial populations in the warm subseafloor of deep-sea hydrothermal vents. Here we reconstruct 73 metagenome-assembled genomes (MAGs) from two geochemically distinct vent fields in the Mid-Cayman Rise to investigate patterns of genomic variation within subseafloor populations. Low-abundance populations with high intra-population diversity coexist alongside high-abundance populations with low genomic diversity, with taxonomic differences in patterns of genomic variation between the mafic Piccard and ultramafic Von Damm vent fields. Populations from Piccard are significantly enriched in nonsynonymous mutations, suggesting stronger purifying selection in Von Damm relative to Piccard. Comparison of nine Sulfurovum MAGs reveals two high-coverage, low-diversity MAGs from Piccard enriched in unique genes related to the cellular membrane, suggesting these populations were subject to distinct evolutionary pressures that may correlate with genes related to nutrient uptake, biofilm formation, or viral invasion. These results are consistent with distinct evolutionary histories between geochemically different vent fields, with implications for understanding evolutionary processes in subseafloor microbial populations.
    Description: R.E.A. was supported by a NASA Postdoctoral Fellowship with the NASA Astrobiology Institute. This work was supported by a NASA Astrobiology Science and Technology for Exploring Planets (ASTEP) grant NNX-327 09AB75G and a grant from Deep Carbon Observatory's Deep Life Initiative to J.A.H. and J.S.S., and the NSF Science and Technology Center for Dark Energy Biosphere Investigations (C-DEBI). Ship and vehicle time in 2012 was supported by the NSF-OCE grant OCE-1061863 to J.S.S.
    Repository Name: Woods Hole Open Access Server
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  • 10
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    Diversity and distribution of subseafloor Thermococcales populations in diffuse hydrothermal vents at an active deep-sea volcano in the northeast Pacific Ocean (2007)
    American Geophysical Union
    Details
    Publication Date: 2022-05-25
    Description: Author Posting. © American Geophysical Union, 2006. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 111 (2006): G04016, doi:10.1029/2005JG000097.
    Description: The presence, diversity, and distribution of a key group of subseafloor archaea, the Thermococcales, was examined in multiple diffuse flow hydrothermal vents at Axial Seamount, an active deep-sea volcano located in the northeast Pacific Ocean. A polymerase chain reaction (PCR) approach was used to determine if this group of subseafloor indicator organisms showed any phylogenetic distribution that may indicate distinct subseafloor communities at vents with different physical and chemical characteristics. Targeted primers for the Thermococcales 16S rRNA (small subunit ribosomal RNA) gene and intergenic transcribed spacer (ITS) region were designed and applied to organisms filtered in-situ directly from a variety of diffuse flow vents. Thermococcales were amplified from 9 of 11 samples examined, and it was determined that the ITS region is a better phylogenetic marker than the 16S rRNA in defining consistent groups of closely related sequences. Results show a relationship between environmental clone distribution and source vent chemistry. The most highly diluted vents with elevated iron and alkalinity contained a distinct group of Thermococcales as defined by the ITS region, suggesting separate subseafloor Thermococcales populations at diffuse vents within the Axial caldera.
    Description: This work was supported by Washington Sea Grant (NA76RG0119), National Science Foundation (OCE 9816491), NSF IGERT (DGE- 9870713), NASA Astrobiology Institute through the Carnegie Geophysical Institute, the NOAA/PMEL Vents Program, NOAA West Coast and Polar Undersea Research Center, and by the Joint Institute for the Study of the Atmosphere and Ocean under NOAA Cooperative Agreement No. NA117RJ1232.
    Keywords: Subseafloor ; Hydrothermal vent ; Intergenic Transcribed Spacer (ITS) region ; Geochemistry ; Diffuse flow vent
    Repository Name: Woods Hole Open Access Server
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