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Identification of Novel Methane-, Ethane-, and Propane-Oxidizing Bacteria at Marine Hydrocarbon Seeps by Stable Isotope Probing

Redmond, Molly C. and Valentine, David L. and Sessions, Alex L. (2010) Identification of Novel Methane-, Ethane-, and Propane-Oxidizing Bacteria at Marine Hydrocarbon Seeps by Stable Isotope Probing. Applied and Environmental Microbiology, 76 (19). pp. 6412-6422. ISSN 0099-2240. PMCID PMC2950463.

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Marine hydrocarbon seeps supply oil and gas to microorganisms in sediments and overlying water. We used stable isotope probing (SIP) to identify aerobic bacteria oxidizing gaseous hydrocarbons in surface sediment from the Coal Oil Point seep field located offshore of Santa Barbara, California. After incubating sediment with ^(13)C-labeled methane, ethane, or propane, we confirmed the incorporation of ^(13)C into fatty acids and DNA. Terminal restriction fragment length polymorphism (T-RFLP) analysis and sequencing of the 16S rRNA and particulate methane monooxygenase (pmoA) genes in ^(13)C-DNA revealed groups of microbes not previously thought to contribute to methane, ethane, or propane oxidation. First, ^(13)C methane was primarily assimilated by Gammaproteobacteria species from the family Methylococcaceae, Gammaproteobacteria related to Methylophaga, and Betaproteobacteria from the family Methylophilaceae. Species of the latter two genera have not been previously shown to oxidize methane and may have been cross-feeding on methanol, but species of both genera were heavily labeled after just 3 days. pmoA sequences were affiliated with species of Methylococcaceae, but most were not closely related to cultured methanotrophs. Second, ^(13)C ethane was consumed by members of a novel group of Methylococcaceae. Growth with ethane as the major carbon source has not previously been observed in members of the Methylococcaceae; a highly divergent pmoA-like gene detected in the ^(13)C-labeled DNA may encode an ethane monooxygenase. Third, ^(13)C propane was consumed by members of a group of unclassified Gammaproteobacteria species not previously linked to propane oxidation. This study identifies several bacterial lineages as participants in the oxidation of gaseous hydrocarbons in marine seeps and supports the idea of an alternate function for some pmoA-like genes.

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Sessions, Alex L.0000-0001-6120-2763
Additional Information:© 2010 American Society for Microbiology. Received 1 February 2010/Accepted 22 July 2010. Published ahead of print on 30 July 2010. We thank Michael Friedrich and Bianca Pommerenke for early assistance with the SIP technique. Heather Coleman, Blair Paul, and Cristoph Pierre collected samples, and Lichun Zhang performed the fatty acid extractions and GC/MS analyses. This work was funded by a National Science Foundation award (OCE-0447395) and a DOE award (DE-NT0005667) to D.L.V. and a National Science Foundation award (EAR-0645502) to A.L.S.
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Department of Energy (DOE)DE-NT0005667
Issue or Number:19
PubMed Central ID:PMC2950463
Record Number:CaltechAUTHORS:20101022-131749813
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Official Citation:Redmond, M. C., D. L. Valentine, et al. (2010). "Identification of Novel Methane-, Ethane-, and Propane-Oxidizing Bacteria at Marine Hydrocarbon Seeps by Stable Isotope Probing." Appl. Environ. Microbiol. 76(19): 6412-6422.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:20486
Deposited By: Tony Diaz
Deposited On:30 Nov 2010 22:41
Last Modified:03 Oct 2019 02:10

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