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Appl. Environ. Microbiol. doi:10.1128/AEM.02222-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Archaeal Community in a Minerotrophic Fen and T-RFLP-Directed Isolation of a Novel Hydrogenotrophic Methanogen

Department of Microbiology, Cornell University, Ithaca, NY 14853, USA; Department of Natural Resources, Cornell University, Ithaca, NY 14853, USA

^* To whom correspondence should be addressed. Email: shz1{at}cornell.edu.

	Abstract

Minerotrophic fen peatlands are widely distributed in northern latitudes and because of their rapid turnover of organic matter, are potentially larger sources of atmospheric methane than bog peatlands per unit area. However, studies on the archaeal community composition in fens are scarce particularly in minerotrophic sites. Several 16S rRNA based primer sets were used to obtain a broad characterization of the archaeal community in a minerotrophic fen in central New York State. A wide archaeal diversity was observed in the site: 11 euryarchaeal and 2 crenarchaeal groups most of which were uncultured. The E1 group, a novel cluster in the Methanomicrobiales order, and Methanosaetaceae were the co-dominant members in all libraries and T-RFLP analysis. Given its abundance and potential hydrogenotrophic methane contribution, the E1 group was targeted for culture attempts with a low-ionic strength medium (PM1). Initial attempts yielded Methanospirillum-dominated cultures. However, by incorporating a T-RFLP analysis as a quick selection tool for treatments and replicates, we were able to select an enrichment dominated by E1. Further dilutions to 10^-9 and tracking with T-RFLP yielded an isolated strain named E1-9c. E1-9c is a novel coccoid hydrogenotrophic, mesophilic, slightly acidophilic methanogen, and is highly sensitive to Na₂S concentrations (requires <0.12 mM for growth). We propose E1-9c as the first representative of a novel genus in the Methanomicrobiales order.