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Applied and Environmental Microbiology, May 2005, p. 2642-2652, Vol. 71, No. 5
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.5.2642-2652.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Methane-Oxidizing Bacteria in a California Upland Grassland Soil: Diversity and Response to Simulated Global Change

Hans-Peter Horz,1,2 Virginia Rich,1,{dagger} Sharon Avrahami,1 and Brendan J. M. Bohannan1*

Department of Biological Sciences, Stanford University, Stanford, California 94305,1 Division of Oral Microbiology and Immunology, RWTH Aachen University Hospital, 52074 Aachen, Germany2

Received 11 August 2004/ Accepted 7 December 2004

We investigated the diversity of methane-oxidizing bacteria (i.e., methanotrophs) in an annual upland grassland in northern California, using comparative sequence analysis of the pmoA gene. In addition to identifying type II methanotrophs commonly found in soils, we discovered three novel pmoA lineages for which no cultivated members have been previously reported. These novel pmoA clades clustered together either with clone sequences related to "RA 14" or "WB5FH-A," which both represent clusters of environmentally retrieved sequences of putative atmospheric methane oxidizers. Conservation of amino acid residues and rates of nonsynonymous versus synonymous nucleotide substitution in these novel lineages suggests that the pmoA genes in these clades code for functionally active methane monooxygenases. The novel clades responded to simulated global changes differently than the type II methanotrophs. We observed that the relative abundance of type II methanotrophs declined in response to increased precipitation and increased atmospheric temperature, with a significant antagonistic interaction between these factors such that the effect of both together was less than that expected from their individual effects. Two of the novel clades were not observed to respond significantly to these environmental changes, while one of the novel clades had an opposite response, increasing in relative abundance in response to increased precipitation and atmospheric temperature, with a significant antagonistic interaction between these factors.

* Corresponding author. Mailing address: Department of Biological Sciences, 371 Serra Mall, Stanford University, Stanford, CA 94305. Phone: (650) 723-3344. Fax: (650) 723-0589. E-mail: bohannan{at}stanford.edu.

{dagger} Present address: MIT/Woods Hole Oceanographic Institution Joint Program in Biological Oceanography, Massachusetts Institute of Technology, Cambridge, MA 02139.

Applied and Environmental Microbiology, May 2005, p. 2642-2652, Vol. 71, No. 5
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.5.2642-2652.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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