免培养法对一热泉细菌多样性的初步研究

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16S rDNA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Thermodesulfobacteria属之间。经PCR扩增出上述5个克隆16S rDNA插入序列中及环境样品总DNA中的16S rDNA V8高变区约600bp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，还显示了它们是该高温热泉中细菌的优势物种。

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

Bacterial biodiversity of the hot spring Dagunguo in Tengchong Rehai in Yunnan Province, 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Hydrogenobacter and Pseudomonas, and the other one was clustered with the Family Thermodesulfobacteriaceae between an unpublished genus Geothermobacterium and Thermodesulfobacteria. The V8 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 PCR products were run DGGE. The pattern of DGGE and the phylogenetic tree of the five 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.