采用PCR-DGGE技术分析蒙古羊瘤胃液相和固相细菌的多样性

Evaluate the diversity of bacteria associated with liquid and solid phases of rumen contents of Mongolian Sheep using PCR-DGGE analysis

为探讨蒙古羊瘤胃液相和固相细菌的多样性,采用变性梯度凝胶电泳技术结合聚合酶链式反应(PCRDGGE)及条带的克隆测序,比较瘤胃液相和固相细菌的差异,同时采用聚类分析和主成分分析(PCA)方法分析瘤胃液相和固相细菌的多样性。结果表明:1)瘤胃液相和固相样品均含有物种丰富的细菌菌群,但液相比固相具有更高的平均条带数,分别为30条和24条;2)瘤胃液相样品比固相样品具有更高的多样性指数、均匀度和丰富度,其数值分别为3.45、0.90和30.20及3.23、0.84和24.62;3)同一动物个体瘤胃液相样品和固相样品聚类在一起,相似性系数均高达0.81;4)共性条带测序结果表明瘤胃的优势细菌主要是Uncultured rumen bacterium和Uncultured Bacteroidetes bacterium,而特异性条带主要是Alcaligenes sp.。不同动物个体的瘤胃液相和固相均含有丰富的细菌菌群,且物种丰富度均较高;同一动物个体的液相与固相相比具有更高的细菌多样性。

This experiment is to explore the diversity of bacteria associated with liquid phase samples and solid phase samples of rumen contents of Mongolian Sheep.This study choice the research methods of polymerase chain reactiondenaturing gradient gel electrophoresis（PCR-DGGE）.The results were analyzed by cluster analysis and PCA.Some common and special bands were identified at the same time.The results showed that both the bacteria from the liquid phase samples and solid phase samples had a high diversity,and the liquid phase samples had higher number of bands compared with the solid phase,were 30 and 24,respectively.Compared with the solid phase samples,the shannon diversity index,evenness and richness of liquid phase samples were higher,were 3.45,0.90,30.20 and 3.23,0.84,24.62,respectively.Samples of rumen liquid phase samples and solid phase samples from the same individual clustered together,and the similarity coefficient were up to 0.81.Uncultured rumen bacterium and Uncultured Bacteroidetes bacterium were predominadt in the rumen of sheep.The special bancterial in rumen content was Alcaligenes sp..Furthermore,both the liquid phase samples and solid phase samples of rumen contents of Mongolian Sheep had abundant bacteria and highest diversity in the liquid phase samples.