西湖底泥中的反硝化型甲烷厌氧氧化菌的分子生物学检测  

Molecular detection of denitrifying anaerobic methane oxidizing bacteria in the sediment of West Lake, Hangzhou

作　　者：朱群[1] 沈李东[2] 胡宝兰[2] 楼莉萍[2] 程东庆[1] 

机构地区：[1]浙江中医药大学生命科学院,杭州310053 [2]浙江大学环境工程系,杭州310058

出　　处：《环境科学学报》2013年第5期1321-1325,共5页Acta Scientiae Circumstantiae

基　　金：国家"十二五"科技支撑计划(No.2012BAJ25B07);国家自然科学基金(No.51108408)~~

摘　　要：反硝化型甲烷厌氧氧化反应(Denitrifying anaerobic methane oxidation,DAMO)是一种最新发现的生物反应,该反应能够偶联反硝化和甲烷的厌氧氧化.催化DAMO反应的微生物是NC10门中一种被命名为"Candidatus Methylomirabilis oxyfera"的细菌.本研究采用基因克隆文库技术考察了西湖淡水底泥中DAMO微生物的分布与种群多样性状况.16S rRNA基因系统发育分析表明,西湖底泥中存在NC10门细菌,与已知的M.oxyfera的16S rRNA基因相似度为93%～98%.DAMO微生物功能基因(pmoA)的系统发育分析进一步证实了西湖底泥中分布有此类微生物,与已知的M.oxyfera的pmoA基因相似度为86%～95%.实时定量PCR结果表明,西湖底泥中DAMO微生物的16S rRNA基因的拷贝数为2.15×105 copies·g-1(以干重计).Denitrifying anaerobic methane oxidation （DAMO）, a coupling process between methane oxidation and denitrification, is discovered recently. This process is mediated by ＂Candidatus Methylomirabilis oxyfera＂, which belongs to the candidate phylum NC10. This study investigated the distribution and diversity of the DAMO bacteria in the sediment of West Lake, Hangzhou. Both the 16S rRNA and the pmoA molecular biomarkers confirmed the presence of diverse NC10 phylum bacteria related to M. oxyfera in this freshwater ecosystem. Phylogenetic analysis showed that the recovered 16S rRNA gene sequences had 93%~98% identity to the 16S rRNA gene of M. oxyfera. The detected pmoA sequences had 86%~95% identity to the pmoA gene of M.oxyfera. Real-time quantitative PCR further confirmed the presence of the DAMO bacteria in the examined sediments with 2.15×10^5 copies·g^-1 （dry weight）.

关 键 词：反硝化型甲烷厌氧氧化菌 西湖底泥 分布 种群多样性 

分 类 号：X172[环境科学与工程—环境科学]