聚合酶链式反应-变性梯度凝胶电泳技术分析1只死亡野生川金丝猴胃肠道菌群多样性  

Bacteria Diversity in Gastrointestinal Tract of a Dead Wild Rhinopithecus roxellanae Analyzed by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis Technology

作　　者：曾燕[1] 牛李丽[2] 邓家波[2] 王强[2] 余建秋[2] 李杨[1] 周梦佳 徐帅[1] 林奕岑 曾东[1] 倪学勤[1] 

机构地区：[1]四川农业大学动物医学院动物微生态研究中心,成都611130 [2]成都动物园,成都610081 [3]四川省畜牧科学研究院,成都610066

出　　处：《动物营养学报》2018年第2期649-657,共9页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：成都大熊猫繁育研究基金会项目(CPF2015-06);国家自然科学基金(31672318)

摘　　要：本试验旨在对1只死亡野生川金丝猴的胃肠道菌群多样性及其克隆测序条带的系统进化树进行分析。取死亡野生川金丝猴胃肠道内容物,进行聚合酶链式反应(PCR)-变性梯度凝胶电泳(DGGE)分析,结合条带的克隆测序、聚类分析和主成分分析(PCA)检测菌群多样性并构建系统进化树。结果显示:1)野生川金丝猴整个胃肠道栖息着大量细菌,且来自胃、小肠的样品聚为一大簇,来自大肠的样品聚为一簇,而来自粪便的样品单独聚为一簇。2)从DGGE图谱上共回收18个条带,细菌种类鉴定主要是5个菌门,分别为变形菌门(Proteobacteria,38.89%)、厚壁菌门(Firmicutes,22.22%)、拟杆菌门(Bacteroidetes,5.56%)、放线菌门(Actinobacteria,5.56%)、疣微菌门(Verrucomicrobia,5.56%)和不可培养菌(uncultured bacterium,22.22%)。其中,变形菌门和厚壁菌门分布于整个胃肠道。3)菌群的系统进化树分析表明,仅有1种不可培养菌与已鉴定的粪肠球菌进化分类相似,而其他不可培养菌与已知的菌种进化分支差异较大,说明在野生川金丝猴的胃肠道中仍有大量的菌群信息未被鉴定。结果提示,本试验测定的1只死亡野生川金丝猴的胃肠道优势菌群为变形菌门,菌群多样性随着胃肠道由前至后的顺序呈现高—低—高的趋势。The aim of this experiment was to evaluate the bacteria diversity in gastrointestinal tract （GIT） of a dead wild Rhinopithecus roxeUanae, and to analyze the phylogenetic tree of cloning sequencing bands. The GIT contents of a dead wild Rhinopithecus roxellanae were collected, and the polymerase chain reaction （PCR） -denaturing gradient gel electrophoresis （DGGE） technology with the cloning sequencing, cluster anal- ysis and principal component analysis （PCA） of bands were used to detect the bacterial diversity and to build the phylogenetic tree. The results showed as follows ~ 1 ） numerous bacteria were obtained from the GIT of wild Rhinopithecus roxellanae. Samples from the stomach, duodenum, jejunum and ileum clustering together, sam- ples from the cecum, colon and rectum clustering together, and samples from the faeces single clustering to- gether. 2） The 18 identified DGGE bands were belong to five phylum, they were Proteobacteria （38.89%）, Firmicutes （ 22.22% ）, Bacteroidetes （ 5.56% ）, Acfinobacteria （ 5.56% ）, Verrucomicrobia （ 5.56% ） and uncultured bacterium （22.22%）. The Proteobacteria and Firmicutes were detected along the GIT. 3） The phy- logenefic tree analysis showed that only one of uncultured bacterium was similar to the identified classification of Enterococcus faecalis, while the other three uncultured bacteria had a significant difference with the known branch of the bacteria. This indicated that a large number of flora information was unkowned in the GIT of the wild Rhinopithecus roxellanae. The results suggest that Proteobacteria is predominant in the GIT of a dead wild Rhinopithecus roxeUanae, and the bacteria diversity shows a tendency of high-low-high according to the GIT from front to back.

关 键 词：川金丝猴 胃肠道菌群 PCR-DGGE 克隆测序 系统进化树 

分 类 号：S826[农业科学—畜牧学]