Genetic basis for nitrate resistance in Desulfovibrio strains

Korte, Hannah L.; Fels, Samuel R.; Christensen, Geoff A.; Price, Morgan N.; Kuehl, Jennifer V.; Zane, Grant M.; Deutschbauer, Adam M.; Arkin, Adam P.; Wall, Judy D.

doi:10.3389/fmicb.2014.00153

Genetic basis for nitrate resistance in Desulfovibrio strains

Journal Article · Sun Apr 20 20:00:00 EDT 2014 · Frontiers in Microbiology

DOI:https://doi.org/10.3389/fmicb.2014.00153· OSTI ID:1628107

Korte, Hannah L. ^[1]; Fels, Samuel R. ^[2]; Christensen, Geoff A. ^[3]; Price, Morgan N. ^[4]; Kuehl, Jennifer V. ^[4]; Zane, Grant M. ^[3]; Deutschbauer, Adam M. ^[4]; Arkin, Adam P. ^[4]; Wall, Judy D. ^[5]

Univ. of Missouri, Columbia, MO (United States). Dept. of Biochemistry; Ecosystems and Networks Integrated with Genes and Molecular Assemblies, Berkeley, CA (United States); DOE/OSTI
Ecosystems and Networks Integrated with Genes and Molecular Assemblies, Berkeley, CA (United States); Univ. of Missouri, Columbia, MO (United States). Dept. of Molecular Microbiology and Immunology
Univ. of Missouri, Columbia, MO (United States). Dept. of Biochemistry; Ecosystems and Networks Integrated with Genes and Molecular Assemblies, Berkeley, CA (United States)
Ecosystems and Networks Integrated with Genes and Molecular Assemblies, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Physical Biosciences Division
Univ. of Missouri, Columbia, MO (United States). Dept. of Biochemistry; Ecosystems and Networks Integrated with Genes and Molecular Assemblies, Berkeley, CA (United States); Univ. of Missouri, Columbia, MO (United States). Dept. of Molecular Microbiology and Immunology

Nitrate is an inhibitor of sulfate-reducing bacteria (SRB). In petroleum production sites, amendments of nitrate and nitrite are used to prevent SRB production of sulfide that causes souring of oil wells. A better understanding of nitrate stress responses in the model SRB, Desulfovibrio vulgaris Hildenborough and Desulfovibrio alaskensis G20, will strengthen predictions of environmental outcomes of nitrate application. Nitrate inhibition of SRB has historically been considered to result from the generation of small amounts of nitrite, to which SRB are quite sensitive. Here we explored the possibility that nitrate might inhibit SRB by a mechanism other than through nitrite inhibition. We found that nitrate-stressed D. vulgaris cultures grown in lactate-sulfate conditions eventually grew in the presence of high concentrations of nitrate, and their resistance continued through several subcultures. Nitrate consumption was not detected over the course of the experiment, suggesting adaptation to nitrate. With high-throughput genetic approaches employing TnLE-seq for D. vulgaris and a pooled mutant library of D. alaskensis, we determined the fitness of many transposon mutants of both organisms in nitrate stress conditions. We found that several mutants, including homologs present in both strains, had a greatly increased ability to grow in the presence of nitrate but not nitrite. The mutated genes conferring nitrate resistance included the gene encoding the putative Rex transcriptional regulator (DVU0916/Dde_2702), as well as a cluster of genes (DVU0251-DVU0245/Dde_0597-Dde_0605) that is poorly annotated. Follow-up studies with individual D. vulgaris transposon and deletion mutants confirmed high-throughput results. We conclude that, in D. vulgaris and D. alaskensis, nitrate resistance in wild-type cultures is likely conferred by spontaneous mutations. Furthermore, the mechanisms that confer nitrate resistance may be different from those that confer nitrite resistance.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1628107

Journal Information:: Frontiers in Microbiology, Journal Name: Frontiers in Microbiology Vol. 5; ISSN 1664-302X

Publisher:: Frontiers Research FoundationCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
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Genetic basis for nitrate resistance in Desulfovibrio strains

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