动物营养学报2018，Vol.30Issue(2)：649-657,9.DOI:10.3969/j.issn.1006-267x.2018.02.030
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
摘要
Abstract
The aim of this experiment was to evaluate the bacteria diversity in gastrointestinal tract (GIT) of a dead wild Rhinopithecus roxellanae,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 analysis 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,samples from the cecum,colon and rectum clustering together,and samples from the faeces single clustering together.2) The 18 identified DGGE bands were belong to five phylum,they were Proteobacteria (38.89％),Firmicutes (22.22％),Bacteroidetes (5.56％),Actinobacteria (5.56％),Verrucomicrobia (5.56％) and uncultured bacterium (22.22％).The Proteobacteria and Firmicutes were detected along the GIT.3) The phylogenetic 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 roxellanae,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Key words
Rhinopithecus roxellanae/gastrointestinal tract bacteria/PCR-DGGE/cloning sequencing/phylogenetic tree分类
农业科技引用本文复制引用
曾燕,曾东,倪学勤,牛李丽,邓家波,王强,余建秋,李杨,周梦佳,徐帅,林奕岑..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[J].动物营养学报,2018,30(2):649-657,9.基金项目
成都大熊猫繁育研究基金会项目(CPF2015-06) （CPF2015-06）
国家自然科学基金(31672318) （31672318）
