Open AccessQuantification of mcrA by fluorescent PCR in methanogenic and methanotrophic microbial communities
Author(s) -
Nunoura Takuro,
Oida Hanako,
Miyazaki Junichi,
Miyashita Ai,
Imachi Hiroyuki,
Takai Ken
Publication year - 2008
Publication title -
fems microbiology ecology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.377
H-Index - 155
eISSN - 1574-6941
pISSN - 0168-6496
DOI - 10.1111/j.1574-6941.2008.00451.x
Subject(s) - biology , 16s ribosomal rna , metagenomics , methanogen , ribosomal rna , archaea , euryarchaeota , gene , primer (cosmetics) , bacteria , microbiology and biotechnology , genetics , chemistry , organic chemistry
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.