动物营养学报 >

2013 , Vol. 25 >Issue 9: 2136 - 2142

DOI: https://doi.org/10.3969/j.issn.1006-267x.2013.09.028

研究简报 Short Communications

采用PCR-DGGE技术分析瘤胃菌群在不同纤维素富集条件下的多样性

  • 曾燕 ,
  • 孙朋 ,
  • 倪学勤 ,
  • 杨杰 ,
  • 曾东 ,
  • 张洪瑜
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  • 1. 四川农业大学动物医学院, 雅安 625014;
    2. 动物疫病与人类健康四川省重点实验室, 雅安 625014

收稿日期: 2013-03-16

  网络出版日期: 2013-08-18

基金资助

教育部留学回国人员科研启动基金资助;教育部长江学者和创新团队发展计划(IRT0848);四川农业大学大学生科研兴趣项目

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Diversity of Rumen Microflora under Different Cellulose Enrichment Conditions Analyzed by PCR-DGGE

  • ZENG Yan ,
  • SUN Peng ,
  • NI Xueqin ,
  • YANG Jie ,
  • ZENG Dong ,
  • ZHANG Hongyu
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  • 1. College of Animal Medicine, Sichuan Agricultural University, Ya'an 625014, China;
    2. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Ya'an 625014, China

Received date: 2013-03-16

  Online published: 2013-08-18

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摘要

本试验旨在采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析瘤胃菌群在不同纤维素富集条件下的多样性。试验采用羧甲基纤维素(CMC)培养基、蛋白胨纤维素(PCS)培养基、J培养基和K培养基分别在37和50℃条件下培养瘤胃内容物,使用PCR-DGGE技术、DGGE图谱共性及特异性条带的克隆和测序、聚类分析和主成分分析(PCA)几种方法分析瘤胃菌群多样性。结果表明:37℃条件下,不同培养基的DGGE图谱差异较大,J培养基和PCS培养基间的相似性系数为0.77;J培养基和CMC培养基间的相似性系数仅为0.76。50℃条件下不同培养基的DGGE图谱相似性较高,J培养基和PCS培养基间的相似性系数为0.78;而CMC培养基和PCS培养基间的相似性系数高达0.84。DGGE图谱中共性条带属于解没食子酸链球菌和解脲芽孢杆菌2个菌属,是瘤胃中的优势菌群;而主要的特异性条带属于解没食子酸链球菌、摩氏假单胞菌、产碱杆菌和未培养毛螺旋菌4个菌属。结果提示,不同纤维素富集培养基和温度对瘤胃菌群的多样性有一定程度上的影响。

关键词： 瘤胃菌群; 纤维素; PCR-DGGE; 聚类分析; 主成分分析

本文引用格式

曾燕 , 孙朋 , 倪学勤 , 杨杰 , 曾东 , 张洪瑜 . 采用PCR-DGGE技术分析瘤胃菌群在不同纤维素富集条件下的多样性[J]. 动物营养学报, 2013 , 25(9) : 2136 -2142 . DOI: 10.3969/j.issn.1006-267x.2013.09.028

Abstract

This experiment was conducted to study the diversity of rumen microflora under different cellulose enrichment conditions by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Carboxymethyl cellulose (CMC), peptone-cellulose (PCS), J and K mediums were used in this study, and rumen contents were incubated at 37 and 50℃, respectively. The diversity of rumen mircroflora was analyzed by PCR-DGGE, cloning and sequencing of common and specific bands in DGGE profiles, cluster analysis and principal component analysis. The results showed as follows: there were differences among DGGE profiles of different mediums at 37℃, the similarity coefficient between J and PCS mediums was 0.77, and that between J and CMC mediums was 0.76. The DGGE profiles of different mediums were similar to each other at 50℃, the similarity coefficient between J and PCS mediums was 0.78, and that between CMC and PCS mediums reached 0.84. The common bands were Streptococcus gallolyticus and Ureibacillus themosphaericus,and were predominant in rumen contents; the specific bands were Streptococcus gallolyticus, Pseudomonas mosselii, Alcaligenes and uncultured Lachnospiraceae bacterium clone. In conclusion, the diversity of rumen microflora can be affected by different cellulose enrichment mediums and culture temperatures.

Key words： rumen microflora; cellulose; PCR-DGGE; cluster analysis; principal component analysis

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