“传统”垃圾填埋场渗滤液中古细菌16S rRNA基因的RFLP分析

Characterization of Archaeal Diversity in the Leachate of a Traditional Landfill by Determining Restriction Fragment Length Polymorphisms of 16S rRNA Genes

“传统”垃圾填埋场渗滤液中古细菌群落的多样性通过不依赖于培养的分析而获得。将渗滤液中古细菌的16S rRNA基因片断(16S rDNA)选择性地扩增出来并用于构建16S rDNA克隆文库。文库内古细菌16S rDNA的遗传变异性通过RFLP分析(限制性内切酶Hha Ⅰ和HaeⅢ)而得到。80个随机选出的古细菌rDNA克隆子被划分为29个不同的RFLP型(组),其中最大的5个型共占所有被分析克隆子的53％左右,而其余24个型的丰度均处于相对较低的水平,其中16个型更仅含有1个克隆子。有关结果表明,对克隆16S rDNA片断的RFLP分析是评估复杂厌氧系统中微生物群落多样性的有力工具。

The diversity of Archaea community associated with the leachate of a traditional municipal solid waste landfill was examined by cultivation-independent analysis. Total-community genomic DNA was isolated from the leachate and archaeal 16S rRNA gene sequences (16S rDNA) were selectively amplified. The 16S rDNA amplicon was subcloned and further characterized by a restriction fragment length polymorphism analysis (RFLP, enzymes Hha Ⅰ and Hae Ⅲ ). A total of 80 cloned 16S rDNA fragments were analyzed, and they were finally clustered into 29 different groups (RFLP patterns), with five most abundant groups accounting for 53% of all the 16S rDNA clones. The remaining 24 groups presented at low levels, of which a total of 16 groups were represented by a single clone. Our results demonstrated that RFLP analysis of PCR-amplified 16S rDNA is a powerful tool for assessing the diversity of microbial communities in complex anaerobic ecosystems.