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

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...

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Bibliographic Details
Published in:Applied and Environmental Microbiology 2010-10, Vol.76 (19), p.6412-6422
Main Authors: Redmond, Molly C, Valentine, David L, Sessions, Alex L
Format: Article
Language:English
Subjects:
Bacteria - classification
Bacteria - genetics
Bacteria - metabolism
Biodiversity
Biological and medical sciences
California
Carbon Isotopes - metabolism
Cluster Analysis
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
DNA, Bacterial - metabolism
DNA, Ribosomal - chemistry
DNA, Ribosomal - genetics
Ethane - metabolism
Fatty acids
Fatty Acids - metabolism
Fundamental and applied biological sciences. Psychology
Geologic Sediments - microbiology
Hydrocarbon leakage & seepage
Hydrocarbons
Isotopes
Metagenome
Methane
Methane - metabolism
Methylococcaceae
Methylophaga
Microbial Ecology
Microbiology
Microorganisms
Molecular Sequence Data
Oxidation
Oxidation-Reduction
Phylogeny
Propane - metabolism
RNA, Ribosomal, 16S - genetics
Sediments
Sequence Analysis, DNA
Staining and Labeling - methods
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Summary: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 ¹³C-labeled methane, ethane, or propane, we confirmed the incorporation of ¹³C 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 ¹³C-DNA revealed groups of microbes not previously thought to contribute to methane, ethane, or propane oxidation. First, ¹³C 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, ¹³C 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 ¹³C-labeled DNA may encode an ethane monooxygenase. Third, ¹³C 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.
ISSN:0099-2240
1098-5336
1098-6596
DOI:10.1128/aem.00271-10