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通过直接检索16S rRNA基因序列研究全尺寸循环填埋场渗滤液中古菌群落的多样性和结构。

Diversity and structure of the archaeal community in the leachate of a full-scale recirculating landfill as examined by direct 16S rRNA gene sequence retrieval.

作者信息

Huang Li-Nan, Zhou Hui, Chen Yue-Qin, Luo Shuo, Lan Chong-Yu, Qu Liang-Hu

机构信息

Key Laboratory of Gene Engineering of the Ministry of Education, School of Life Sciences, Zhongshan University, Guangzhou 510275, PR China.

出版信息

FEMS Microbiol Lett. 2002 Sep 10;214(2):235-40. doi: 10.1111/j.1574-6968.2002.tb11353.x.

DOI:10.1111/j.1574-6968.2002.tb11353.x
PMID:12351237
Abstract

The diversity and structure of the archaeal community in the effluent leachate from a full-scale recirculating landfill was characterized by direct 16S rRNA gene (16S rDNA) retrieval. Total-community DNA was extracted from the microbial assemblages in the landfill leachate, and archaeal 16S rDNAs were amplified with a universally conserved primer and an Archaea-specific primer. The amplification product was then used to construct a 16S rDNA clone library, and 70 randomly selected archaeal clones in the library were grouped by restriction fragment length polymorphism (RFLP) analysis. Sequencing and phylogenetic analysis of representatives from each unique RFLP type showed that the archaeal library was dominated by methanogen-like rDNAs. Represented in the kingdom of Euryarchaeota were phylotypes highly similar to the methanogenic genera Methanoculleus, Methanosarcina, Methanocorpusculum, Methanospirillum and Methanogenium, where the clone distribution was 48, 11, 3, 1 and 1, respectively. No sequences related to known Methanosaeta spp. were retrieved. Four rDNA clones were not affiliated with the known methanogenic Archaea, but instead, they were clustered with the uncultured archaeal sequences recently recovered from anaerobic habitats. Two chimeric sequences were identified among the clones analyzed.

摘要

通过直接检索16S rRNA基因(16S rDNA)对一座大型循环式垃圾填埋场渗滤液中古菌群落的多样性和结构进行了表征。从垃圾填埋场渗滤液中的微生物群落中提取总群落DNA,并用通用保守引物和古菌特异性引物扩增古菌16S rDNA。然后将扩增产物用于构建16S rDNA克隆文库,并通过限制性片段长度多态性(RFLP)分析对文库中随机选择的70个古菌克隆进行分组。对每种独特RFLP类型的代表进行测序和系统发育分析表明,古菌文库以类产甲烷菌的rDNA为主。广古菌门中的代表菌群与产甲烷菌属Methanoculleus、Methanosarcina、Methanocorpusculum、Methanospirillum和Methanogenium高度相似,其中克隆分布分别为48、11、3、1和1。未检索到与已知甲烷八叠球菌属物种相关的序列。四个rDNA克隆与已知的产甲烷古菌无关,而是与最近从厌氧生境中获得的未培养古菌序列聚类。在所分析的克隆中鉴定出两个嵌合序列。

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