Open AccessDevelopment of 16S rRNA gene‐targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs)
Author(s) -
Miyashita Ai,
Mochimaru Hanako,
Kazama Hiromi,
Ohashi Akiyoshi,
Yamaguchi Takashi,
Nunoura Takuro,
Horikoshi Koki,
Takai Ken,
Imachi Hiroyuki
Publication year - 2009
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2009.01648.x
Subject(s) - 16s ribosomal rna , biology , ribosomal rna , archaea , euryarchaeota , microbiology and biotechnology , gene , genetics
Uncultured archaeal anaerobic methanotrophs (ANMEs) are known to operate the anaerobic oxidation of methane process, an important sink for the greenhouse gas methane in natural environments. In this study, we designed 16S rRNA gene‐specific primers for each of the phylogenetic groups of ANMEs (ANME‐1, Guaymas Basin hydrothermal sediment clones group within the ANME‐1, ANME‐2a, ANME‐2b, ANME‐2c and ANME‐3) based on previously reported sequences. The newly designed primers were used for the detection of the various groups of ANMEs in the sulphate‐limited anaerobic environmental samples, i.e. methanogenic sludges, rice field soils, lotus field sediments and natural gas fields. The ANME 16S rRNA gene sequences were detected only in a natural gas field sample among the environments examined in this study and were of the ANME‐1 and ‐2c groups. In addition, the quantitative real‐time PCR analysis using the designed primers showed that abundances of ANME‐1 and ‐2c were estimated to be <0.02% of the total prokaryotic 16S rRNA gene community. The newly designed ANME group‐specific primers in this study may be useful to survey the distribution and quantitative determination of ANMEs.