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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

Samantha A. Morris,1 Stefan Radajewski,1 Toby W. Willison,2 and J. Colin Murrell1*

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.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom. Phone: 44 (024) 76-523-553. Fax: 44 (024) 76-523-568. E-mail: cmurrell{at}bio.warwick.ac.uk.

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.



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