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Analysis of methanotrophic bacteria in Movile Cave by stable isotope probing

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UNSPECIFIED. (2004) Analysis of methanotrophic bacteria in Movile Cave by stable isotope probing. ENVIRONMENTAL MICROBIOLOGY, 6 (2). pp. 111-120. ISSN 1462-2912

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Official URL: http://dx.doi.org/10.1046/j.1462-2920.2003.00543.x

Abstract

Movile Cave is an unusual groundwater ecosystem that is supported by in situ chemoautotrophic production. The cave atmosphere contains 1-2% methane (CH4), although much higher concentrations are found in gas bubbles that keep microbial mats afloat on the water surface. As previous analyses of stable carbon isotope ratios have suggested that methane oxidation occurs in this environment, we hypothesized that aerobic methane-oxidizing bacteria (methanotrophs) are active in Movile Cave. To identify the active methanotrophs in the water and mat material from Movile Cave, a microcosm was incubated with a 10%(CH4)-C-13 headspace in a DNA-based stable isotope probing (DNA-SIP) experiment. Using improved centrifugation conditions, a C-13-labelled DNA fraction was collected and used as a template for polymerase chain reaction amplification. Analysis of genes encoding the small-subunit rRNA and key enzymes in the methane oxidation pathway of methanotrophs identified that strains of Methylomonas, Methylococcus and Methylocystis/Methylosinus had assimilated the (CH4)-C-13, and that these methanotrophs contain genes encoding both known types of methane monooxygenase (MMO). Sequences of non-methanotrophic bacteria and an alga provided evidence for turnover of CH4 due to possible cross-feeding on C-13-labelled metabolites or biomass. Our results suggest that aerobic methanotrophs actively convert CH4 into complex organic compounds in Movile Cave and thus help to sustain a diverse community of microorganisms in this closed ecosystem.

Item Type: Journal Article
Subjects: Q Science > QR Microbiology
Journal or Publication Title: ENVIRONMENTAL MICROBIOLOGY
Publisher: BLACKWELL PUBLISHING LTD
ISSN: 1462-2912
Date: February 2004
Volume: 6
Number: 2
Number of Pages: 10
Page Range: pp. 111-120
Identification Number: 10.1046/j.1462-2920.2003.00543.x
Publication Status: Published
URI: http://wrap.warwick.ac.uk/id/eprint/8838

Data sourced from Thomson Reuters' Web of Knowledge

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