《中国畜牧兽医》---唯一指定的官方网站 ›› 2015, Vol. 42 ›› Issue (6): 1402-1408.doi: 10.16431/j.cnki.1671-7236.2015.06.012

• 生物技术 • 上一篇    下一篇

圈养老年大熊猫肠道菌群16SrDNA克隆文库的建立

王晓艳1,3, 袁听1,3, 廖虹2, 罗永久3, 何廷美4, 王承东4, 刘小敏4, 吴虹林4, 彭广能3   

  1. 1. 重庆三峡职业学院动物科技系, 重庆 404100;
    2. 雅安市城乡规划建设和住房保障局, 雅安 625014;
    3. 四川农业大学动物医学院, 动物疫病与人类健康四川省重点实验室, 雅安 625014;
    4. 中国林业科学研究院 大熊猫保护研究中心, 四川雅安碧峰峡基地, 雅安 625007
  • 收稿日期:2014-12-15 出版日期:2015-06-20 发布日期:2015-07-23
  • 通讯作者: 彭广能 E-mail:pgn.sicau@163.com
  • 作者简介:王晓艳(1988-),女,四川泸州人,硕士,研究方向:兽医临床疾病的诊断与治疗,E-mail:wxycqsxedu@163.com 袁 听(1988-),男,四川雅安人,硕士,研究方向:小动物临床疾病的诊断与治疗,E-mail:51198773@qq.com 廖 虹(1966-),女,四川富顺人,学士,研究方向:动物福利与保护,E-mail:1276473089@qq.com
  • 基金资助:

    国家自然基金面上项目(31272620);大熊猫国际合作资金项目(SD0632);教育部"长江学者和创新团队发展规划"创新团队项目(IRT0848)

Construction of the 16S rDNA Clone Library of Intestinal Flora of the Captive Elderly Giant Panda

WANG Xiao-yan1,3, YUAN Ting1,3, LIAO Hong2, LUO Yong-jiu3, HE Ting-mei4, WANG Cheng-dong4, LIU Xiao-min4, WU Hong-lin4, PENG Guang-neng3   

  1. 1. Department of Animal Science and Technology, Chongqing Three Gorges Vocational College, Chongqing 404100, China;
    2. Ya'an City Planning and Construction and Housing Security Bureau, Ya'an 625014, China;
    3. Key Laboratory of Animal Disease and Human Health of Sichuan Province, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China;
    4. Sichuan Ya'an Bifengxia Base, Giant Panda Research Center, Chinese Academy of Forestry Science, Ya'an 625007, China
  • Received:2014-12-15 Online:2015-06-20 Published:2015-07-23

摘要: 为了研究老年大熊猫肠道菌群的构成,本试验对3只圈养老年大熊猫粪便细菌构建16S rDNA克隆文库,采用限制性内切酶Hinf Ⅰ、Msp Ⅰ对其进行限制性片段长度多态性(restriction fragment length polymorphism,RFLP)及测序分析.结果显示,老年大熊猫肠道细菌主要由变形菌门和厚壁菌门组成;变形菌门中又以大肠埃希氏菌属为主,其次为假单胞菌属、志贺氏菌属、气单胞菌属;而厚壁菌门中以链球菌属为主,其次为魏斯氏菌属、梭菌属;此外还发现一定比例的未培养细菌.本试验第一次建立了较丰富的老年大熊猫肠道菌群的克隆文库,为分析比较各年龄层大熊猫的肠道菌群结构的异同提供了参照,也为合理饲喂老年大熊猫,保障老年大熊猫的健康提供重要信息.

关键词: 圈养; 老年大熊猫; 肠道菌群; 16S rDNA克隆文库

Abstract: In order to comparatively analyze the differences and similarities of intestinal flora from all ages of giant pandas,we built 16S rDNA clone library of three captive elderly giant pandas'fecal bacteria and made restriction fragment length polymorphism (RFLP) and sequencing analysis by using restriction enzymes HinfⅠ and MspⅠ.The results showed that the intestinal flora of elderly giant panda were mainly attributable to two bacteria:Proteobacteria and Firmicutes.Between them,the main in Proteobacteria of intestinal flora of elderly giant panda were Escherichia coli,followed by Pseudomonas sp.,Shigella and Aeromonas;The main in Firmicutes were Streptococcus,followed by Weissella,Clostridium;Of which there were a certain proportion of bacteria were uncultured bacteria.The article firstly established a rich clone library of intestinal flora from elderly giant pandas,which provided a reference to comparatively analyze the similarities and differences in structure of intestinal flora from all ages of giant pandas,as well as provided the important information about reasonable feeding elderly giant pandas and protecting health of elderly giant pandas.

Key words: captivity; elderly giant panda; intestinal flora; 16S rDNA clone library

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