Publication Date:
2022-10-26
Description:
© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in D’Angelo, T., Goordial, J., Poulton, N., Seyler, L., Huber, J., Stepanauskas, R., & Orcutt, B. Oceanic crustal fluid single cell genomics complements metagenomic and metatranscriptomic surveys with orders of magnitude less sample volume. Frontiers in Microbiology, 12, (2022): 738231, https://doi.org/10.3389/fmicb.2021.738231.
Description:
Fluids circulating through oceanic crust play important roles in global biogeochemical cycling mediated by their microbial inhabitants, but studying these sites is challenged by sampling logistics and low biomass. Borehole observatories installed at the North Pond study site on the western flank of the Mid-Atlantic Ridge have enabled investigation of the microbial biosphere in cold, oxygenated basaltic oceanic crust. Here we test a methodology that applies redox-sensitive fluorescent molecules for flow cytometric sorting of cells for single cell genomic sequencing from small volumes of low biomass (approximately 103 cells ml–1) crustal fluid. We compare the resulting genomic data to a recently published paired metagenomic and metatranscriptomic analysis from the same site. Even with low coverage genome sequencing, sorting cells from less than one milliliter of crustal fluid results in similar interpretation of dominant taxa and functional profiles as compared to ‘omics analysis that typically filter orders of magnitude more fluid volume. The diverse community dominated by Gammaproteobacteria, Bacteroidetes, Desulfobacterota, Alphaproteobacteria, and Zetaproteobacteria, had evidence of autotrophy and heterotrophy, a variety of nitrogen and sulfur cycling metabolisms, and motility. Together, results indicate fluorescence activated cell sorting methodology is a powerful addition to the toolbox for the study of low biomass systems or at sites where only small sample volumes are available for analysis.
Description:
The borehole observatories that form the backbone of this project were funded by the Integrated Ocean Drilling Program (IODP, now the International Ocean Discovery Program), the United States National Science Foundation (NSF), and the Gordon and Betty Moore Foundation (grant GBMF1609). Cruise AT39-01 was funded by the NSF (OCE-1634025 to C. Geoff Wheat). Analyses were funded by the NSF (OCE-1536623 to BO; OIA-1826734 to RS, NP, and BO; and OCE-16435208 and OCE-1745589 to JH), the NSF-funded Center for Dark Energy Biosphere Investigations (C-DEBI) Science and Technology Center (via subawards from OIA-0939564 to BO and JH), and the NASA Exobiology program (80NSSC19K0466 to BO). This is C-DEBI publication 571.
Keywords:
Deep biosphere
;
Oceanic crust
;
Crustal fluid
;
Single cell genomics
;
Metatranscriptomics
;
IODP
;
CORKS
;
North Pond
Repository Name:
Woods Hole Open Access Server
Type:
Article
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