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

作　　者：王涛[1] 柴丽红[1] 崔晓龙[1] 彭谦[1] 姜成林[1] 

机构地区：[1]云南大学云南省微生物研究所教育部微生物资源开放重点实验室,昆明650091

出　　处：《微生物学报》2003年第5期541-546,共6页Acta Microbiologica Sinica

基　　金：国家自然科学基金资助项目 (3 9960 0 0 3 );科技部基础研究重大项目前期研究专项(2 0 0 1CCC0 0 60 0 );云南省教育厅 2 0 0 1年度教育基金 (0 11114 2 )~~

摘　　要：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and Pseudomonas, and the other one was clustered with the Family Thermode sulf obacteriaceae between an unpublished genus Geothermobacterium and Thermode sulfobacteria.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.

关 键 词：免培养法 热泉 细菌多样性 16SrDNA 

分 类 号：Q938[生物学—微生物学]