艾丁湖沉积物放线菌多样性  

Studies on the Actinobacterial Diversity in Aiding Lake Sediments

作　　者：夏占峰[1,3] 关统伟[1,3] 阮继生[2] 黄英[2] 张利莉[1,3] 

机构地区：[1]塔里木大学,新疆生产建设兵团塔里木盆地生物资源保护利用重点实验室,阿拉尔843300 [2]中国科学院微生物研究所,微生物资源前期开发国家重点实验室,北京100101 [3]塔里木大学生命科学学院,阿拉尔843300

出　　处：《微生物学报》2011年第8期1023-1031,共9页Acta Microbiologica Sinica

基　　金：国家“973计划”前期研究专项(2010CB134505);国家自然科学基金项目(31060003);教育部科学技术重点项目(209145);中国科学院微生物资源前期开发国家重点实验室开放课题(SKLMR-20090603)~~

摘　　要：【目的】为了研究新疆艾丁湖低海拔、高盐环境中的放线菌多样性。【方法】本研究应用基于16S rRNA基因序列系统发育分析的免培养方法和选择性分离培养方法相结合的方式对艾丁湖沉积物样品进行放线菌多样性分析。利用放线菌特异性引物扩增16S rRNA基因并构建了16S rRNA基因克隆文库,对文库中的插入序列进行RFLP(Restriction Fragment Length Polymorphism限制性片段长度多态性)分析。采用不同盐浓度的9种选择性培养基分离样品中的放线菌。【结果】通过对16S rRNA基因克隆文库分析,273个克隆分布于放线菌纲(Actinobacteria)的放线菌亚纲(Actinobacteridae 208个)、酸微菌亚纲(Acidimicrobidae 13个)和红色杆菌亚纲(Rubrobacteridae 52个)。该环境中优势放线菌为微球菌科的罗斯氏菌属(Rothia),占到克隆总数的37%。有45.8%的序列与已有效发表菌株的序列相似性小于97%,部分序列形成了几个独立的进化分支,可能代表着放线菌新的类群。通过分离培养,共得到55株放线菌,分布在放线菌目的6个亚目中,以链霉菌属(Streptomyces)和拟诺卡氏菌属(Nocardiopsis)为主要类群。有6株菌的16S rRNA基因序列与已有效发表菌株的序列相似性小于97%,为潜在的放线菌新分类单元。【结论】研究表明,艾丁湖中的放线菌具有较高的多样性,并蕴藏着多种新类型,有进一步研究价值。[Objective] This study aims at investigating the diversity of actinobacteria in Aiding Lake,a hypersaline lake and the lowest land point in China.[Methods] The diversity of actinobacteria in the sediment from Aiding Lake was investigated by culture-independent method based on phylogenetic analysis of 16S rRNA gene sequences and selective isolation.Specific primers were used to amplify the actinobacterial 16S rRNA gene,and corresponding clone libraries were constructed for the sediment samples.Different clones selected on the basis of HaeⅢ digestion patterns were sequenced.Nine selective media with different salinities were used to isolate actinobacteria from the sediment samples.[Results] The analysis of 16S rRNA gene sequences showed that 273 clone sequences belonged to subclasses Actinobacteridae（208）,Acidimicrobidae（13） and Rubrobacteridae（52）.The dominant actinobacteria was genus Rothia,which accounted for 37% of total clones.The similarity between 45.8% of 273 detected sequences and published sequences were less than 97%,which might represent new taxa.Some sequences,which formed several distinct clades in phylogenetic tree may represent new taxonomical groups of actinobacteria.Fifty-five strains were isolated by different selective media.They belonged to six suborders of the order Actinomycetales,of which Streptomyces and Nocardiopsis were the dominant groups.Six potential new species were obtained.[Conclusion] Aiding Lake harbors abundant actinobacteria,including large number of unknown actinobacterial groups.

关 键 词：艾丁湖 放线菌多样性 免培养方法 选择性分离 

分 类 号：Q938.8[生物学—微生物学]