沁水盆地寺河地区煤层水细菌多样性分析

Bacteria diversity in coalbed water at Sihe area of Qinshui Basin

[目的]了解沁水盆地寺河地区煤层水中细菌群落组成和物种多样性。[方法]采用免培养法提取煤层水中微生物总DNA,利用细菌通用引物构建16S r DNA基因克隆文库。采用HhaⅠ、MspⅠ限制性内切酶对克隆子进行RFLP分析,测序并构建16S r DNA基因系统发育树。[结果]从文库中筛选出234个阳性克隆,覆盖度为97.4%,聚类为28个操作分类单元。BLAST比对、RDP归类及系统发育分析将这234个克隆归为变形菌门、拟杆菌门、螺旋体门、疣微菌门、黏胶球形菌门。其中变形菌门为绝对优势类群,占整个细菌克隆文库的80.1%。变形菌门中的ε-变形菌纲,占整个基因文库的35%。[结论]应用16S r DNA克隆文库技术,分析沁水盆地寺河地区煤层水中细菌类群的多样性不高。

[ Objective] In order to investigate bacteria composition and diversity in coalbed water at Sihe area of Qinshui Ba- sin,Shanxi. [ Methods] Environmental total DNA was directly extracted from combed water. The 16S rDNA genes were ampli- fied from the DNA by PCR with bacteria - specific primers and were constructed the clone library. The gene - containing clones were digested with restriction endonucleases Hha I and Msp I for restriction fragment length polymorphism（RFLP） analysis. The unique RFLP clones were sequenced and constructed phylogenetic tree. [ Results ] In total, 234 clones were screened and grouped into 28 operational taxonomic units（OTUs）. The clone coverage C value was 97.4%. The 234 clones were divided into 5 phyla（ Proteobacteria, Bacteroidetes, Spirochaetes, Verrucomicrobia, and Lentisphaerae） with BLAST analysis, RDP classifer and phylogenetic a^alysis. Proteobacteria was the absolutely dominant group and accounted 80.1 percent of the clone library. And s - Proteobacteria of Proteobacteria was the first major group with 35% proportion of the whole clone library. [ Conclusion] Bacterial diversity in coalbed water at Sihe area of Qinshui Basin was low using 16S rDNA clone library analysis.