不同纤维素培养基及温度对体外培养瘤胃细菌多样性的影响  

Evaluate the diversity of rumen bacteria on anaerobic culture with different cellulose enrichment medium and temperature in vitro

作　　者：曾燕[1] 孙朋[1] 倪学勤[1,2] 杨杰[1] 曾东[1,2] 张洪瑜[1,2] 

机构地区：[1]四川农业大学动物医学院,四川雅安625014 [2]动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国农业大学学报》2013年第6期148-152,共5页Journal of China Agricultural University

基　　金：教育部留学回国人员科研启动基金资助(200835-1);四川省学术带头人培养基金资助

摘　　要：为探讨瘤胃细菌多样性,采用聚合酶链式反应-变性梯度凝胶电泳(Polymerase chain reaction-denaturing gradient gel electrophoresis,PCR-DGGE)技术结合共性及特异性条带的克隆测序和聚类分析及主成分分析,对牛体外厌氧条件下瘤胃细菌在CMC、PCS、J、K培养基及37和50℃培养温度下的多样性进行研究。结果表明:1)37℃条件下不同培养基的DGGE图谱差异较大,虽然具有较高的平均条带数,CMC和K培养基样品分别为20和27条,但各样品间的相似性系数较低,J和CMC培养基间的相似性系数为0.84,J和PCS培养基间的相似性系数仅为0.75。2)50℃条件下不同培养基的DGGE图谱差异较小,平均条带数较低,CMC和K培养基样品均为15条,但各样品间的相似性较高,CMC和PCS培养基间的相似性系数为0.89;而J和PCS培养基间的相似性系数高达0.97。3)DGGE图谱中共性条带序列表明瘤胃优势细菌主要是Streptococcus macedonicus,而特异性条带主要是Escherichia coli、Firmicutes以及Brevundimonas。体外厌氧条件下不同纤维素培养基和温度在一定程度上影响瘤胃细菌的多样性。The study is to explore the diversity of rumen bacteria of anaerobic culture in vitro. The research methods included polymerase chain reaction-denaturing gradient gel electrophoresis（PCR-DGGE）, cluster analysis and PCA. CMC,PCS,J and K mediums were used in this trial and three rumen content samples were incubated at 37 and 50 ℃ respectively. Some common and special bands were identified at the same time. The results showed that the DGGE profiles of different mediums were significantly to each other at 37 ℃, although it had the higher number of bands which the average bands of CMC and k mediums were 20 and 27. The similarity coefficient between J and CMC medium was 0.84, and it reduced to 0.75 between J and PCS medium. The DGGE profiles of different mediums were similar to each other at 50 ℃ ,it had the lower number of bands,and the average samples of CMC and k mediums were only with 15 bands. However, the coefficient between CMC and PCS medium was 0.89, while the similarity coefficient between J and PCS medium could get 0.97. Streptococcus macedonicus was predominant in the upper G1 tract. The special bands in rumen content were Escherichia coil, Firmicutes and Brevundimonas. Furthermore, the diversity of rumen bacterial of anaerobic culture to some extent was affected by cellulose enrichment conditions in vitro.

关 键 词：PCR—DGGE 瘤胃细菌 纤维素 聚类分析 主成分分析(PCA) 

分 类 号：S182[农业科学—农业基础科学]