免培养法研究野生川金丝猴肠道内生细菌多样性  

Culture-independent analysis of intestinal microbial diversity from Rhinopithecus roxellana

作　　者：杨曼[1] 兰阿峰[1,2] 郭素芬[1] 丁小维[1,2] 邓百万[1,2] 

机构地区：[1]陕西理工学院生物科学与工程学院,陕西汉中723001 [2]陕西省食药用菌工程技术研究中心,陕西汉中723001

出　　处：《微生物学通报》2014年第8期1605-1612,共8页Microbiology China

基　　金：陕西省科技厅科技统筹创新计划项目(No.2012HBGC-20);陕西理工学院人才引进启动项目

摘　　要：【目的】了解野生川金丝猴(Rhinopithecus roxellana)肠道内生细菌的组成及其多样性。【方法】提取川金丝猴肠道内生细菌总DNA,选用细菌通用引物799F和1492R对总DNA进行16S rRNA基因特异性扩增,构建川金丝猴肠道内生细菌16S rRNA基因克隆文库,对阳性克隆进行限制性内切酶片段长度多态性(PCR-RFLP)分析,并对HaeⅢ酶切带谱菌株进行测序,构建系统发育树。【结果】根据酶切带谱分析和测序结果,将随机挑取的157个阳性克隆归为27个不同的可操作分类单元(OTUs)。系统发育分析表明这些克隆序列有62.10%属于厚壁菌门(Firmicutes),其中包括梭菌属(Clostridium)、Cellulosilyticum属、Robinsoniella属、Anaerofustis属、Blautia属和Anaerovorax属,有37.90%属于未培养细菌。【结论】川金丝猴肠道内生细菌多样性丰富,并且可能存在新的分类单元。[Objective] The composition and diversity of intestinal microbial diversity of wild Si- chuan snub-nosed monkey （Rkinopitkecus roxellana） were investigated. [Methods] The total DNA extracted from endophytic bacterial was performed 16S rRNA gene specific amplification by used bacteria-specific primers 799F and 1492R and constructed a clone library. Then, 16S rRNA gene phylogenetic tree was established by analyzed positive clones used polymerase chain reac- tion-restriction fragment length polymorphism （PCR-RFLP） and by sequenced some of the clones. [Results] The 157 clones analyzed were grouped into 27 operational taxonomic units （OTUs）, where the clone distribution was as follows： 62.10% of Firmicutes which contained Clostridium, Cellulosilyticum, Robinsoniella, Anaerofustis, Anaerovorax and Blautia, 37.90% of uncultured bacteria. [Conclusion] The intestine bacterial of wild Sichuan snub-nosed monkey has a rich di- versity and maybe exist new taxon.

关 键 词：免培养 川金丝猴 肠道内生细菌 多样性 

分 类 号：S865.3[农业科学—野生动物驯养] Q93[农业科学—畜牧兽医]