Apportioning bacterial carbon source utilization in soil using 14C isotope analysis of FISH-targeted bacterial populations sorted by Fluorescence Activated Cell Sorting (FACS): 14C-FISH-FACS

Gougoulias, C., Meade, A. ORCID: https://orcid.org/0000-0001-7095-7711 and Shaw, L. (2018) Apportioning bacterial carbon source utilization in soil using 14C isotope analysis of FISH-targeted bacterial populations sorted by Fluorescence Activated Cell Sorting (FACS): 14C-FISH-FACS. Environmental Microbiology Reports, 10 (3). pp. 245-254. ISSN 1758-2229 doi: 10.1111/1758-2229.12631

Abstract/Summary

An unresolved need in microbial ecology is methodology to enable quantitative analysis of in situ microbial substrate carbon use at the population level. Here, we evaluated if a novel combination of radiocarbon- labelled substrate tracing, Fluorescence in situ Hybridisation (FISH) and Fluorescence-Activated Cell Sorting (FACS) to sort the FISH-targeted population for quantification of incorporated radioactivity (14C-FISH-FACS) can address this need. Our test scenario used FISH probe PSE1284 targeting Pseudomonas spp. (and some Burkholderia spp.) and salicylic acid added to rhizosphere soil. We examined salicylic acid-14C fate (mineralized, cell-incorporated, extractable and non-extractable) and mass balance (0-24 h) and show that the PSE1284 population captured ~50% of the Nycodenz extracted biomass 14C. Analysis of the taxonomic distribution of the salicylic acid biodegradation trait suggested that PSE1284 population success was not due to conservation of this trait but due to competitiveness for the added carbon. Adding 50KBq of 14C sample-1 enabled detection of 14C in the sorted population at ~60-600 times background; a sensitivity which demonstrates potential extension to analysis of rarer/ less active populations. Given its sensitivity and compatibility with obtaining a C mass balance, 14C-FISH-FACS allows quantitative dissection of C flow within the microbial biomass that has hitherto not been achieved.

Item Type	Article
URI	https://reading-pure-test.eprints-hosting.org/id/eprint/75599
Identification Number/DOI	10.1111/1758-2229.12631
Refereed	Yes
Divisions	Life Sciences > School of Biological Sciences > Ecology and Evolutionary Biology Science > School of Archaeology, Geography and Environmental Science > Department of Geography and Environmental Science Central Services
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