Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs)
Development of 16S rRNA gene-targeted primers for detection of archaeal anaerobic methanotrophs (ANMEs)
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DOI：
10.1111/j.1574-6968.2009.01648.x
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发表时间：
2009-08-01
影响因子：
2.1
通讯作者：
Imachi, Hiroyuki
中科院分区：
文献类型：
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作者：
Miyashita, Ai;Mochimaru, Hanako;Imachi, Hiroyuki
未培养的古菌厌氧甲烷氧化菌（ANME）是甲烷厌氧氧化过程中的重要生物，是自然环境中温室气体甲烷的重要汇。在本研究中，我们设计了16 S rRNA基因特异性引物的ANME-1，瓜伊马斯盆地热液沉积物克隆组内的ANME-1，ANME-2a，ANME-2b，ANME-2c和ANME-3的系统发育组的基础上，以前报道的序列。将新设计的引物用于检测硫酸盐限制的厌氧环境样品（即产甲烷污泥、稻田土壤、莲田沉积物和天然气田）中的各种ANME。ANME 16 S rRNA基因序列仅在天然气田样品中检测到，在这项研究中所研究的环境中，ANME-1和-2c组。此外，利用所设计的引物进行的实时荧光定量PCR分析表明，ANME-1和ANME-2c的丰度估计为< 0.02%的总原核16 S rRNA基因社区。本研究设计的ANME组特异性引物可用于ANME的分布和定量检测。
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.