新疆玛纳斯热气泉免培养土壤细菌多样性分析  

Bacterial diversity in a hot gas spring soil of MaNasi County,Xinjiang by culture-independent approach

作　　者：徐建华[1,2] 杨红梅[2] 曾军[1,2] 李智[1,2] 娄恺[2] 

机构地区：[1]新疆大学生命科学与技术学院,乌鲁木齐830046 [2]新疆农业科学院微生物应用研究所,乌鲁木齐830091

出　　处：《微生物学报》2010年第5期568-573,共6页Acta Microbiologica Sinica

基　　金：国家"973项目"前期研究专项(2008CB417214);新疆特殊环境微生物实验室开放课题(XJYS0203-2007-04)~~

摘　　要：【目的】了解新疆玛纳斯热气泉土壤免培养细菌群落组成及多样性。【方法】采用免培养法直接从土壤样品中提取总DNA,利用细菌通用引物对土壤总DNA进行16S rDNA扩增,构建细菌16SrDNA文库。使用HaeⅢ限制性内切酶对阳性克隆进行限制性片段长度多态性分析(restriction fragment length polymorphism,RFLP),挑选具有不同酶切图谱的克隆进行测序、比对并构建16SrDNA系统发育树。【结果】从土壤细菌16S rDNA文库中随机挑选了170个阳性克隆,共得到29个不同的分类单元(operational taxonomic unit,OTU)。系统发育分析归为6个门:酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)和浮霉菌门(Planctomycetes)。其中厚壁菌门(Firmicutes)为绝对优势类群,占整个细菌文库的71%。29个OTUs中有14条序列与GenBank中相关序列的相似性低于97%(序列长度约1.5kb),占序列总数的48%。【结论】热气泉土壤细菌种群多样性较低,但存在大量潜在细菌新种。[Objective]In order to investigate bacteria diversity in a hot gas spring soil of MaNasi county,Xinjiang.[Methods]Total DNA were directly extracted from the soil of a hot gas spring from Xinjiang.16S rDNA were amplified directly from purified DNA by PCR with universally bacteria-specific rDNA primers and cloned.Positive clones were identified by restriction fragment length polymorphism（RFLP）,and unique rDNA types clones were sequenced,analysed and then constructed phylogenetic tree.[Results]Twenty-nine operational taxonomic units （OTU） from 170 positive clones were found belonging to 6 phyla：Firmicutes,Proteobacteria,Acidobacteria,Bacteroidetes,Planctomycetes,Actinobacteria.Firmicutes （71%） was the absolutely dominant components of the soil bacterial community.14 sequences （about 1.5 kb） from 29 OTUs showed less affiliation with known taxa（97% sequence similarity）.[Conclusion]Soil bacterial diversity is low in the hot gas spring soil,but exit a large number of new unknown taxon in this environment.

关 键 词：细菌多样性 16SrDNA 热气泉 地下煤火 限制性片段长度多态性分析(restriction FRAGMENT length polymorphism RFLP) 

分 类 号：Q938[生物学—微生物学]