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  • 1
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    Massachusetts Institute of Technology and Woods Hole Oceanographic Institution
    Publication Date: 2022-05-25
    Description: Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the Massachusetts Institute of Technology and the Woods Hole Oceanographic Institution February 2001
    Description: Processes that enable marine phytoplankton to acquire trace metals are fundamental to our understanding of primary productivity and global carbon cycling. This thesis explored the biogeochemistry of cobalt using analytical chemistry and physiological experiments with the dominant phytoplankton species, Prochlorococcus. A high sensitivity method for Co speciation was developed using hanging mercury drop cathodic stripping voltammetry. Dissolved Co at the Bermuda Atlantic Time Series station (BATS) in the Sargasso Sea was bound by strong organic complexes with a conditional stability constant of logK=16.3l0.9. A depth profile of Co at BATS revealed a nutrient-like profile. Biweekly time series measurements of total cobalt near Bermuda from the MITESS sampler were 0-47pM throughout 1999, and averaged 20±10pM in 1999. A transect of total cobalt from BATS to American coastal waters ranged from 19- 133pM and correlated negatively with salinity (r2=0.93), suggestive of coastal waters as an input source. Prochlorococcus strains MED4-Ax and SS120 showed an absolute requirement for Co, despite replete Zn. 57Co uptake rates and growth rates were enhanced by additions of filtered low Co cultures, suggesting that a ligand is present that facilitates Co uptake. Bottle incubations from a Synechococcus bloom in the Pacific showed production of 425pM strong cobalt ligand. These and other lines of evidence support the hypothesis that a cobalt ligand, or cobalophore, is involved in cobalt uptake. Co-limited Prochlorococcus cultures exhibited an increase in the fraction of cells in G2 relative to other cell cycle stages during exponential growth, and the durations of this stage increased with decreasing cobalt concentrations. This effect was not observed with Fe, N, or P-limited cultures, suggestive of a specific biochemical function of cobalt that would interfere with the late stages of the cell cycle. The ligand Teta was explored as a means to induce cobalt limitation. The CoTeta complex was not bioavailable to the Sargasso Sea microbial assemblage in short-term experiments. Bottle incubations with Teta did not induce cobalt limitation of Prochlorococcus. These results are consistent with the lower conditional stability constant for CoTeta (logK=11.2l0.1) relative to natural cobalt ligands in seawater, and with culture studies that suggest uptake of cobalt via strong organic ligands.
    Description: The work in this thesis was supported by a grant from the National Science Foundation (#OCE-9618729) for cyanobacteria metal interactions in the Sargasso Sea. I have been funded through WHOI on an NSF coastal traineeship (#DGE-9454129) for my first year, followed by an EP A STAR Graduate Fellowship for the subsequent years. Additional funding was supplied by the WHOI Educational Endowment Funds and by the WHOI Ditty Bag fund for part of the DNA/cell cycle work.
    Keywords: Biogeochemistry ; Cobalt ; Marine phytoplankton ; Cyanobacteria ; Oceanus (Ship : 1975-) Cruise OC349
    Repository Name: Woods Hole Open Access Server
    Type: Thesis
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  • 2
    Publication Date: 2022-05-25
    Description: Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of National Academy of Sciences for personal use, not for redistribution. The definitive version was published in Proceedings of the National Academy of Sciences 108 (2011): 4352-4357, doi:10.1073/pnas.1016106108.
    Description: Harmful algal blooms (HABs) cause significant economic and ecological damage worldwide. Despite considerable efforts, a comprehensive understanding of the factors that promote these blooms has been lacking because the biochemical pathways that facilitate their dominance relative to other phytoplankton within specific environments have not been identified. Here, biogeochemical measurements demonstrated that the harmful 43 Aureococcus anophagefferens outcompeted co-occurring phytoplankton in estuaries with elevated levels of dissolved organic matter and turbidity and low levels of dissolved inorganic nitrogen. We subsequently sequenced the first HAB genome (A. anophagefferens) and compared its gene complement to those of six competing phytoplankton species identified via metaproteomics. Using an ecogenomic approach, we specifically focused on the gene sets that may facilitate dominance within the environmental conditions present during blooms. A. anophagefferens possesses a larger genome (56 mbp) and more genes involved in light harvesting, organic carbon and nitrogen utilization, and encoding selenium- and metal-requiring enzymes than competing phytoplankton. Genes for the synthesis of microbial deterrents likely permit the proliferation of this species with reduced mortality losses during blooms. Collectively, these findings suggest that anthropogenic activities resulting in elevated levels of turbidity, organic matter, and metals have opened a niche within coastal ecosystems that ideally suits the unique genetic capacity of A. anophagefferens and thus has facilitated the proliferation of this and potentially other HABs.
    Description: Joint Genome Institute is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231. Efforts were also supported by awards from New York Sea Grant to Stony Brook University, National Oceanic and Atmospheric Administration Center for Sponsored Coastal Ocean Research award #NA09NOS4780206 to Woods Hole Oceanographic Institution, NIH grant GM061603 to Harvard University, and NSF award IOS-0841918 to The University of Tennessee.
    Keywords: Harmful algal blooms ; HABs ; Genome sequence ; Ecogenomics ; Metaproteomics ; Eutrophication ; Aureococcus anophagefferens
    Repository Name: Woods Hole Open Access Server
    Type: Preprint
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  • 3
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    Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu
    Publication Date: 2022-10-31
    Description: Dataset: Trichodesmiium field metaproteomes - sample provenance
    Description: Trichodesmium sample provenance collected in North Atlantic surface waters, at station BATS (Bermuda Atlantic Time-series Study), and station ALOHA (A Long-Term Oligotrophic Habitat Assessment) between 2000 and 2018. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/787093
    Description: Gordon and Betty Moore Foundation (GBMF) GBMF3934, Gordon and Betty Moore Foundation: Marine Microbiology Initiative (MMI) GBMF3782, NSF Division of Ocean Sciences (NSF OCE) OCE-1657766, NSF Division of Ocean Sciences (NSF OCE) OCE-1850719
    Keywords: Trichodesmium ; Metaproteomics
    Repository Name: Woods Hole Open Access Server
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  • 4
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    Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu
    Publication Date: 2022-10-31
    Description: Dataset: Trichodesmium field metaproteomes - protein spectral counts in alternative format
    Description: Metaproteomes of Trichodesmium from samples collected in North Atlantic surface waters, at station BATS (Bermuda Atlantic Time-series Study), and station ALOHA (A Long-Term Oligotrophic Habitat Assessment) between 2000 and 2018. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/787078
    Description: Gordon and Betty Moore Foundation (GBMF) GBMF3934, Gordon and Betty Moore Foundation: Marine Microbiology Initiative (MMI) GBMF3782, NSF Division of Ocean Sciences (NSF OCE) OCE-1657766, NSF Division of Ocean Sciences (NSF OCE) OCE-1850719
    Keywords: Trichodesmium ; Metaproteomics
    Repository Name: Woods Hole Open Access Server
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  • 5
    Publication Date: 2022-10-31
    Description: Dataset: Trichodesmium field metaproteomes - sequence fasta
    Description: FASTA file of sequences in Trichodesmium field metaproteomes analyzed by 2D LC-MS/MS mapped to a Trichodesmium metagenome (IMG ID 2821474806) plus cyanoGEBA species genomes (Shih et al, 2013). Samples were collected in North Atlantic surface waters, at station BATS (Bermuda Atlantic Time-series Study), and station ALOHA (A Long-Term Oligotrophic Habitat Assessment) between 2000 and 2018. Related datasets: Trichodesmium field metaproteomes - peptide spectral counts: https://www.bco-dmo.org/dataset/787168 Trichodesmium field metaproteomes - protein spectral counts: https://www.bco-dmo.org/dataset/787147 Trichodesmium sample provenance: https://www.bco-dmo.org/dataset/787093 - Sample provenance file, which includes sample locations, filter sizes For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/787181
    Description: Gordon and Betty Moore Foundation (GBMF) GBMF3934, Gordon and Betty Moore Foundation: Marine Microbiology Initiative (MMI) GBMF3782, NSF Division of Ocean Sciences (NSF OCE) OCE-1657766, NSF Division of Ocean Sciences (NSF OCE) OCE-1850719
    Keywords: Trichodesmium ; Metaproteomics
    Repository Name: Woods Hole Open Access Server
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  • 6
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    Biological and Chemical Oceanography Data Management Office (BCO-DMO). Contact: bco-dmo-data@whoi.edu
    Publication Date: 2022-10-31
    Description: Dataset: Trichodesmium field metaproteomes - single colony metaproteomes
    Description: Single colony metaproteomes of Trichodesmium from samples collected in North Atlantic surface waters during the R/V Atlantis cruise AT39-05 in March of 2018. For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/786694
    Description: Gordon and Betty Moore Foundation (GBMF) GBMF3934, Gordon and Betty Moore Foundation: Marine Microbiology Initiative (MMI) GBMF3782, NSF Division of Ocean Sciences (NSF OCE) OCE-1657766, NSF Division of Ocean Sciences (NSF OCE) OCE-1850719
    Keywords: Trichodesmium ; Metaproteomics
    Repository Name: Woods Hole Open Access Server
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  • 7
    Publication Date: 2022-05-26
    Description: © The Author(s), 2011. This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited. The definitive version was published in Frontiers in Microbiology 2 (2011): 215, doi:10.3389/fmicb.2011.00215.
    Description: Improvements in temporal and spatial sampling frequency have the potential to open new windows into the understanding of marine microbial dynamics. In recent years, efforts have been made to allow automated samplers to collect microbial biomass for DNA/RNA analyses from moored observatories and autonomous underwater vehicles. Measurements of microbial proteins are also of significant interest given their biogeochemical importance as enzymes that catalyze reactions and transporters that interface with the environment. We examined the influence of five preservatives solutions (SDS-extraction buffer, ethanol, trichloroacetic acid, B-PER, and RNAlater) on the proteome integrity of the marine cyanobacterium Synechococcus WH8102 after 4 weeks of storage at room temperature. Four approaches were used to assess degradation: total protein recovery, band integrity on an SDS detergent polyacrylamide electrophoresis (SDS-PAGE) gel, and number of protein identifications and relative abundances by 1-dimensional LC–MS/MS proteomic analyses. Total protein recoveries from the preserved samples were lower than the frozen control due to processing losses, which could be corrected for with internal standardization. The trichloroacetic acid preserved sample showed significant loss of protein band integrity on the SDS-PAGE gel. The RNAlater preserved sample showed the highest number of protein identifications (103% relative to the control; 520 ± 31 identifications in RNAlater versus 504 ± 4 in the control), equivalent to the frozen control. Relative abundances of individual proteins in the RNAlater treatment were quite similar to that of the frozen control (average ratio of 1.01 ± 0.27 for the 50 most abundant proteins), while the SDS-extraction buffer, ethanol, and B-PER all showed significant decreases in both number of identifications and relative abundances of individual proteins. Based on these findings, RNAlater was an effective proteome preservative, although further study is warranted on additional marine microbes.
    Description: This work was funded by the National Science Foundation Chemical and Biological Oceanography, Center for Microbial Oceanography Research and Education (C-MORE), and the Gordon and Betty Moore Foundation.
    Keywords: Proteome ; Preservation ; Autonomous sampling ; Cyanobacteria ; Alkaline phosphatase ; Proteomics ; Synechococcus WH8102
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 8
    Publication Date: 2023-03-06
    Description: Dataset: ProteOMZ Exclusive Peptide Level Spectral Counts
    Description: Relative protein abundance from scaled and corrected exclusive peptide spectral counts from 20-1250 m in the water column (0.2-3 µm filter size fraction) from the ProteOMZ R/V Falkor expedition. There are a total of 107,579 unique peptide sequences from 56,543 protein groups (88,251 proteins). Exclusive spectral counts are provided per sample as are the full dataset scaled and normalized spectral counts. The protein distributions in this dataset highlight the microbial dynamics across biomes in the central Pacific Ocean. These data were submitted in Saunders et al. (2022). For a complete list of measurements, refer to the full dataset description in the supplemental file 'Dataset_description.pdf'. The most current version of this dataset is available at: https://www.bco-dmo.org/dataset/868030
    Description: Gordon and Betty Moore Foundation: Marine Microbiology Initiative (MMI) GBMF3782, Schmidt Ocean Institute (SOI) R/V Falkor 160115 SOI ProteOMZ Expedition
    Keywords: Metaproteomics ; Mesopelagic ; Pelagic ; Nitrification ; Methylotrophy
    Repository Name: Woods Hole Open Access Server
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  • 9
    Publication Date: 2023-03-08
    Description: © The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Saunders, J. K., McIlvin, M. R., Dupont, C. L., Kaul, D., Moran, D. M., Horner, T., Laperriere, S. M., Webb, E. A., Bosak, T., Santoro, A. E., & Saito, M. A. Microbial functional diversity across biogeochemical provinces in the central Pacific Ocean. Proceedings of the National Academy of Sciences of the United States of America, 119(37),(2022): e2200014119, https://doi.org/10.1073/pnas.2200014119.
    Description: Enzymes catalyze key reactions within Earth’s life-sustaining biogeochemical cycles. Here, we use metaproteomics to examine the enzymatic capabilities of the microbial community (0.2 to 3 µm) along a 5,000-km-long, 1-km-deep transect in the central Pacific Ocean. Eighty-five percent of total protein abundance was of bacterial origin, with Archaea contributing 1.6%. Over 2,000 functional KEGG Ontology (KO) groups were identified, yet only 25 KO groups contributed over half of the protein abundance, simultaneously indicating abundant key functions and a long tail of diverse functions. Vertical attenuation of individual proteins displayed stratification of nutrient transport, carbon utilization, and environmental stress. The microbial community also varied along horizontal scales, shaped by environmental features specific to the oligotrophic North Pacific Subtropical Gyre, the oxygen-depleted Eastern Tropical North Pacific, and nutrient-rich equatorial upwelling. Some of the most abundant proteins were associated with nitrification and C1 metabolisms, with observed interactions between these pathways. The oxidoreductases nitrite oxidoreductase (NxrAB), nitrite reductase (NirK), ammonia monooxygenase (AmoABC), manganese oxidase (MnxG), formate dehydrogenase (FdoGH and FDH), and carbon monoxide dehydrogenase (CoxLM) displayed distributions indicative of biogeochemical status such as oxidative or nutritional stress, with the potential to be more sensitive than chemical sensors. Enzymes that mediate transformations of atmospheric gases like CO, CO2, NO, methanethiol, and methylamines were most abundant in the upwelling region. We identified hot spots of biochemical transformation in the central Pacific Ocean, highlighted previously understudied metabolic pathways in the environment, and provided rich empirical data for biogeochemical models critical for forecasting ecosystem response to climate change.
    Description: Funding for this research was provided by the Gordon and Betty Moore Foundation (grants 3782 and 8453), the US NSF (NSF grants OCE-1924554, 2123055, 2125063, 2048774, and 2026933), the Center for Chemical Currencies on a Microbial Planet (NSF grant OCE-2019589), and the US NIH General Medicine (grant GM135709-01A1). J.K.S. was supported by a NASA Postdoctoral Program Fellowship with the NASA Astrobiology Program, administered by Universities Space Research Association under contract with NASA. A.E.S. was supported by the Sloan Foundation, the Simons Foundation, and NSF grant OCE-1437310. A portion of this research used resources at the US Department of Energy JGI sponsored by the Office of Biological and Environmental Research and operated under contract DE-AC02-05CH11231 (JGI). C.L.D. and D.K. were supported by NSF grants OCE-1558453 and OCE-2049299. T.H. was supported by NSF grant OCE-2023456.
    Keywords: Marine microbial ecology ; Metaproteomics ; Mesopelagic ; Nitrification ; Methylotrophy
    Repository Name: Woods Hole Open Access Server
    Type: Article
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