免培养法对一热泉细菌多样性的初步研究

Preliminary Study on Bacterial Diversity in a Hot Spring by Using Culture-independent Approach

应用免培养法 (Culture independent)对云南腾冲热海大滚锅高温热泉中细菌的多样性进行初步的分析。经过克隆筛选 ,测定了 5个克隆的 1 6SrDNA插入片段的近全序列 ,系统发育分析的结果表明 ,它们分属于Bacillus、Hydrogenobacter和Pseudomonas,有一个克隆尚难确定其分类地位 ,它属于Thermodesulfobacteriaceae科 ,介于Geothermbacterium属和Thermodesulfo bacteria属之间。经PCR扩增出上述 5个克隆 1 6SrDNA插入序列中及环境样品总DNA中的1 6SrDNAV8高变区约 60 0bp片段 ,进行变性梯度电泳 (DGGE)。所得电泳图谱和 5个序列的系统发育树不仅表明该高温热泉存在着丰富的细菌多样性 。

Bacterial biodiversity of the hot spring Dagunguo in Tengchong Rehai in Yunnan P rovince, China was preliminarily studied with culture-independent method. Five complete 16S rDNA sequences were obtained and phylogenetically analysed. Four of these sequences were closely related to the genera: Bacillus、Hydrogenobacter and Pseudomonas, and the other one was clustered with the Family Thermode sulf obacteriaceae between an unpublished genus Geothermobacterium and Thermode sulfobacteria. The V 8 high variable region(about 600bp) of 16S rDNA from the five 16S rDNA sequences and the genomic DNA were obtained by PCR. All of the PC R products were run DGGE. The pattern of DGGE and the phylogenetic tree of the f ive sequences not only indicated the hot spring Dagunguo of Yunnan province has plenty of bacterial species but also showed they were dominant bacterial species in the hot spring.