Sangam: A Confluence of Knowledge Streams

High-throughput isolation and ecogenomics of phages infecting fastidious marine heterotrophic bacteria

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dc.contributor Temperton, Ben
dc.contributor Allen, Michael
dc.creator Buchholz, H
dc.date 2022-05-31T07:56:30Z
dc.date 2022-05-16
dc.date 2022-05-24T20:51:05Z
dc.date 2022-05-31T07:56:30Z
dc.date.accessioned 2023-02-23T12:14:03Z
dc.date.available 2023-02-23T12:14:03Z
dc.identifier http://hdl.handle.net/10871/129782
dc.identifier.uri http://localhost:8080/xmlui/handle/CUHPOERS/258518
dc.description Marine Bacteria and their associated viruses are key players in shaping microbial community structures and global biogeochemical cycles. Cell lysis through viral predation is a crucial component for the recycling of carbon compounds and other nutrients, and interaction between viruses and hosts can also alter cellular functions via metabolic ‘hijacking’. The culture-based study of phototrophic cyanobacterial virus-host systems has revealed that these constant co29 evolutionary pressures in virus-host systems are further escalated by virally mediate horizontal gene transfer. Viruses of fastidious, heterotrophic bacteria are among the most abundant and ecologically significant virus-host systems in the oceans, but the dearth of cultured model systems has hampered the progression in testing hypotheses and interpretation of meta’omics data. The central goal of this thesis was to establish efficient and high-throughput methodologies for viral isolation, specifically targeting fastidious heterotrophic microbes. Using a novel isolate of the ubiquitous SAR11 clade as a model for highly abundant hosts, and three novel isolates of the methylotrophic OM43 clade for a lower-abundance host, I devised an optimised workflow based on established Dilution-to-Extinction culturing techniques that resulted in the isolation of over 117 viruses. The results show that among these novel viruses were the first known siphoviruses of the SAR11 clade and the first recorded viruses infecting members of the OM43 clade. Genomic evidence by the Methylophilales phage Melnitz indicated that inter-class host transitioning between streamlined heterotrophs may occur, and highlighted unusual viral features such as curli genes and glutamine riboswitches. Furthermore, utilising the power of metagenomics coupled with culture based host range experiments, the Pelagibacter phage Skadi was revealed as a highly abundant polar virus, and as an example for ecotypic niche specificity among the most abundant viruses on Earth. Culture-based identification of virus-host pairs combined with ecogenomic interpretation of metagenomic data will improve our understanding of ecological patterns and of viral strategies. The results from this thesis illuminated parts of the viral “dark matter” and highlighted possible novel viral strategies. This work can be adapted to many different systems, both of high and low abundance, and should be used to improve future viral isolation campaigns.
dc.publisher University of Exeter
dc.publisher Biological Sciences
dc.rights 2022-12-17
dc.rights The information and data given in Chapter 4 is meant for publication in a scientific journal (Likely the ISME journal) within the next couple months. The conclusions drawn in this chapter could be re-constructed from public datasets if given access to our analysis methods and findings, therefore it is critical for us to withhold publication of the thesis until we can finalize the paper manuscript and submit it for publication in a suitable journal. Typically the publication process will take at least a couple months as well, during this period we need the data to be confidential as well. The embargo is therefore strictly necessary to avoid potential conflict.
dc.rights http://www.rioxx.net/licenses/all-rights-reserved
dc.subject Marine Microbiology
dc.subject OM43
dc.subject SAR11
dc.subject Virology
dc.title High-throughput isolation and ecogenomics of phages infecting fastidious marine heterotrophic bacteria
dc.type Thesis or dissertation
dc.type Doctor of Philosophy in Biological Sciences
dc.type Doctoral
dc.type Doctoral Thesis


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Appendix 6 Supplementary Video 2.1.mp4 3.835Mb application/octet-stream View/Open
Appendix 7 tRNAs.xlsx 180.2Kb application/octet-stream View/Open
Appendix 8 ANI pairings.xlsx 23.10Kb application/octet-stream View/Open
BuchholzH.pdf 18.78Mb application/pdf View/Open

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