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Applied and Environmental Microbiology, March 2002, p. 1446-1453, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1446-1453.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification of the Functionally Active Methanotroph Population in a Peat Soil Microcosm by Stable-Isotope Probing

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL,¹ Department of Agriculture, Environment, and Fisheries, Edinburgh EH14 1TY, United Kingdom²

Received 17 August 2001/ Accepted 11 December 2001

The active population of low-affinity methanotrophs in a peat soil microcosm was characterized by stable-isotope probing. "Heavy" ¹³C-labeled DNA, produced after microbial growth on ¹³CH₄, was separated from naturally abundant ¹²C-DNA by cesium chloride density gradient centrifugation and used as a template for the PCR. Amplification products of 16S rRNA genes and pmoA, mxaF, and mmoX, which encode key enzymes in the CH₄ oxidation pathway, were analyzed. Sequences related to extant type I and type II methanotrophs were identified, indicating that these methanotrophs were active in peat exposed to 8% (vol/vol) CH₄. The ¹³C-DNA libraries also contained clones that were related to ß-subclass Proteobacteria, suggesting that novel groups of bacteria may also be involved in CH₄ cycling in this soil.

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