Wang Shenghui, Liu Yanjun, Liu Guofu, Huang Yaru, Zhou Yu
College of Life Science, Liaocheng University, Liaocheng, 252059, People's Republic of China.
Curr Microbiol. 2017 Aug;74(8):908-914. doi: 10.1007/s00284-017-1260-8. Epub 2017 May 13.
Nitrite-dependent anaerobic methane oxidation (n-damo) is catalyzed by the NC10 phylum bacterium "Candidatus Methylomirabilis oxyfera" (M. oxyfera). Generally, the pmoA gene is applied as a functional marker to test and identify NC10-like bacteria. However, it is difficult to detect the NC10 bacteria from sediments of freshwater lake (Dongchang Lake and Dongping Lake) with the previous pmoA gene primer sets. In this work, a new primer cmo208 was designed and used to amplify pmoA gene of NC10-like bacteria. A newly nested PCR approach was performed using the new primer cmo208 and the previous primers cmo182, cmo682, and cmo568 to detect the NC10 bacteria. The obtained pmoA gene sequences exhibited 85-92% nucleotide identity and 95-97% amino acid sequence identity to pmoA gene of M. oxyfera. The obtained diversity of pmoA gene sequences coincided well with the diversity of 16S rRNA sequences. These results indicated that the newly designed pmoA primer cmo208 could give one more option to detect NC10 bacteria from different environmental samples.
依赖亚硝酸盐的厌氧甲烷氧化（n-damo）由NC10门细菌“食烷菌属候选嗜氧菌”（M. oxyfera）催化。通常，pmoA基因用作功能标记来检测和鉴定类NC10细菌。然而，使用先前的pmoA基因引物组很难从淡水湖（东昌湖和东平湖）沉积物中检测到NC10细菌。在这项工作中，设计了一种新引物cmo208，并用于扩增类NC10细菌的pmoA基因。使用新引物cmo208和先前的引物cmo182、cmo682和cmo568进行了一种新的巢式PCR方法来检测NC10细菌。获得的pmoA基因序列与M. oxyfera的pmoA基因具有85-92%的核苷酸同一性和95-97%的氨基酸序列同一性。获得的pmoA基因序列多样性与16S rRNA序列多样性高度吻合。这些结果表明，新设计的pmoA引物cmo208为从不同环境样品中检测NC10细菌提供了更多选择。
