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Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs).
FEMS Microbiol Lett. 2009 Aug; 297(1):31-7.FM

Abstract

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.

Authors+Show Affiliations

Subsurface Geobiology Advanced Research Team, Extremobiosphere Research Program, Institute of Biogeosciences, Japan Agency for Marine-Earth Science and Technology, Kanagawa, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19486160

Citation

Miyashita, Ai, et al. "Development of 16S rRNA Gene-targeted Primers for Detection of Archaeal Anaerobic Methanotrophs (ANMEs)." FEMS Microbiology Letters, vol. 297, no. 1, 2009, pp. 31-7.
Miyashita A, Mochimaru H, Kazama H, et al. Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs). FEMS Microbiol Lett. 2009;297(1):31-7.
Miyashita, A., Mochimaru, H., Kazama, H., Ohashi, A., Yamaguchi, T., Nunoura, T., Horikoshi, K., Takai, K., & Imachi, H. (2009). Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs). FEMS Microbiology Letters, 297(1), 31-7. https://doi.org/10.1111/j.1574-6968.2009.01648.x
Miyashita A, et al. Development of 16S rRNA Gene-targeted Primers for Detection of Archaeal Anaerobic Methanotrophs (ANMEs). FEMS Microbiol Lett. 2009;297(1):31-7. PubMed PMID: 19486160.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs). AU - Miyashita,Ai, AU - Mochimaru,Hanako, AU - Kazama,Hiromi, AU - Ohashi,Akiyoshi, AU - Yamaguchi,Takashi, AU - Nunoura,Takuro, AU - Horikoshi,Koki, AU - Takai,Ken, AU - Imachi,Hiroyuki, Y1 - 2009/05/09/ PY - 2009/6/3/entrez PY - 2009/6/3/pubmed PY - 2010/1/12/medline SP - 31 EP - 7 JF - FEMS microbiology letters JO - FEMS Microbiol. Lett. VL - 297 IS - 1 N2 - 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. SN - 1574-6968 UR - https://www.unboundmedicine.com/medline/citation/19486160/Development_of_16S_rRNA_gene_targeted_primers_for_detection_of_archaeal_anaerobic_methanotrophs__ANMEs__ L2 - https://academic.oup.com/femsle/article-lookup/doi/10.1111/j.1574-6968.2009.01648.x DB - PRIME DP - Unbound Medicine ER -