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Quantification of mcrA by fluorescent PCR in methanogenic and methanotrophic microbial communities

  1. Takuro Nunoura1,
  2. Hanako Oida1,
  3. Junichi Miyazaki1,
  4. Ai Miyashita2,
  5. Hiroyuki Imachi1,
  6. Ken Takai1

Article first published online: 3 MAR 2008

DOI: 10.1111/j.1574-6941.2008.00451.x

Issue

FEMS Microbiology Ecology

FEMS Microbiology Ecology

Volume 64, Issue 2, pages 240–247, May 2008

Additional Information

How to Cite

Nunoura, T., Oida, H., Miyazaki, J., Miyashita, A., Imachi, H. and Takai, K. (2008), Quantification of mcrA by fluorescent PCR in methanogenic and methanotrophic microbial communities. FEMS Microbiology Ecology, 64: 240–247. doi: 10.1111/j.1574-6941.2008.00451.x

Author Information

  1. 1

    Subground Animalcule Retrieval (SUGAR) Program, Extremobiosphere Research Center, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), Yokosuka Japan

  2. 2

    Department of Environmental Systems Engineering, Nagaoka University of Technology, Nagaoka, Japan

*Correspondence: Takuro Nunoura, Subground Animalcule Retrieval (SUGAR) Program, Extremobiosphere Research Center, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, Japan. Tel.: +81 46 867 9707; fax: +81 46 867 9715; e-mail: takuron@jamstec.go.jp

  1. Editor: Gary King

Publication History

  1. Issue published online: 3 MAR 2008
  2. Article first published online: 3 MAR 2008
  3. Received 8 August 2007; revised 25 October 2007; accepted 29 December 2007.First published online 3 March 2008.

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Keywords:

  • methanogen;
  • anaerobic methanotroph;
  • methanogenesis;
  • anaerobic oxidation of methane;
  • mcrA;
  • quantitative fluorescent PCR

Abstract

A quantitative fluorogenic PCR method for detecting methanogenic and methanotrophic orders was established using a refined primer set for the methyl coenzyme M reductase subunit A gene (mcrA). The method developed was applied to several microbial communities in which diversity and abundance of methanogens or anaerobic methanotrophs (ANMEs) was identified by 16S rRNA gene clone analysis, and strong correlations between the copy numbers of mcrA with those of archaeal 16S rRNA genes in the communities were observed. The assay can be applied to detecting and assessing the abundance of methanogens and/or ANMEs in anoxic environments that could not be detected by 16S rRNA gene sequence analyses.

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