研究简报 Short Communications

应用扩增性rDNA限制性酶切片段多态性分析技术研究竹鼠盲肠细菌菌群的多样性

  • 周建波 ,
  • 姜亚 ,
  • 宁俊平 ,
  • 曹晓燕 ,
  • 黄怡 ,
  • 王士长
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  • 广西大学动物科学技术学院, 南宁 530005

收稿日期: 2013-04-11

  网络出版日期: 2013-09-16

基金资助

中柬越英国际合作项目(DelPHE ROUND 5,DelPHE project 852)

Diversity of Cecal Bacterial Flora in Bamboo Rat (Rhizomyidae) Analyzed by Amplified Ribosomal DNA Restriction Analysis Technology

  • ZHOU Jianbo ,
  • JIANG Ya ,
  • NING Junping ,
  • CAO Xiaoyan ,
  • HUANG Yi ,
  • WANG Shichang
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  • Animal Science and Technology of Guangxi University, Nanning 530005, China

Received date: 2013-04-11

  Online published: 2013-09-16

摘要

本试验应用扩增性rDNA限制性酶切片段多态性分析(ARDRA)技术研究竹鼠盲肠细菌菌群的多样性。利用免培养的分子生物学技术,从竹鼠盲肠内容物中提取细菌的总DNA,用细菌通用引物F27/R1492扩增出细菌16S rDNA的基因,构建16S rDNA基因文库。采用HaeⅢ和HhaⅠ 2种限制性内切酶对阳性克隆子的扩增产物进行酶切,挑选不同的操作分类单元(OTU)测序,通过Blast在线比对,分析竹鼠盲肠细菌菌群的多样性。结果表明:竹鼠盲肠中的细菌菌群大部分是未培养或未知的菌株,在分类学上具有潜在意义的被选菌株,而且存在可能和脂肪代谢有关的微生物群体。同时也有乳酸菌属、芽孢杆菌属、梭形杆菌属和多黏类芽孢杆菌属的分布。从结果中看出,芽孢杆菌属、梭形杆菌属和2个未知菌株的克隆数都大于20个,是优势菌株。

关键词: 竹鼠; 16S rDNA; ARDRA; 克隆

本文引用格式

周建波 , 姜亚 , 宁俊平 , 曹晓燕 , 黄怡 , 王士长 . 应用扩增性rDNA限制性酶切片段多态性分析技术研究竹鼠盲肠细菌菌群的多样性[J]. 动物营养学报, 2013 , 25(10) : 2504 -2508 . DOI: 10.3969/j.issn.1006-267x.2013.10.035

Abstract

This experiment was conducted to study the diversity of cecal bacterial flora in bamboo rat (Rhizomyidae) analyzed by amplified ribosomal DNA restriction analysis (ARDRA) technology. The culture-free technology of molecular biology was used to extract microbial total DNA from the cecum of bamboo rat. The bacteria universal primers F27/R1492 were used to amplify bacteria genes of 16S rDNA so as to build up gene library of 16S rDNA. Two kinds of restriction enzymes, HaeⅢ and HhaⅠ, were used to digest amplification products of positive clones, and then different operational taxonomic units (OTU) sequencing were picked up. Through Blast comparing online, the diversity of cecal bacterial flora in bamboo rat was analyzed. The results showed that the most microbes in cecum of bamboo rat were uncultured or unknown strains which were the selected strains of potential significance in taxonomy, and there were microorganism groups related to lipid metabolism. Meanwhile, the distribution of Lactobacillus, Bacillus, Fusiformis and Paenibacillus polymyxa also existed. In conclusion, Bacillus, Fusiformis and two unknown strains whose clone numbers are more than 20 are dominant strains.

Key words: bamboo rat; 16S rDNA; ARDRA; clone

参考文献

[1] VANEECHOUTTE M,ROSSAU R,DE VOS P,et al.Rapid identification of bacteria of the Comamonadaceae with amplified ribosomal DNA-restriction analysis (ARDRA)[J].FEMS Microbiology Letters,1992,93(3):227-234.  
[2] WEIDNER S,ARNOLD W,PUHLER A.Diversity of uncultured microorganisms associated with the seagrass Halophila stipulacea estimated by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes[J].Applied and Environmental Microbiology,1996,62(3):766-771.
[3] HEYNDRICKX M,VANDAMME P,VANDAMME P,et al.Applicability of combined amplified ribosomal DNA restriction analysis (ARDRA) patterns in bacterial phylogeny and taxonomy[J].Journal of Microbiological Methods,1996,26(3):247-259.  
[4] LAGUERRE G,ALLARD M R,REVOY F,et al.Rapid identification of rhizobia by restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes[J].Applied and Environmental Microbiology,1994,60(1):56-63.
[5] ZHOU J Z,BRUNS M A,TIEDJE J M.DNA recovery from soils of diverse composition[J].Applied and Environmental Microbiology,1996,62(2):316-322.
[6] MURRAY A E,PRESTON C M,MASSAWA R,et al.Seasonal and spatial variability of bacterial and archaeal assemblages in the coastal waters near Anvers Island,Antarctica[J].Applied and Environmental Microbiology,1998,64:2585-2595.
[7] LIU B,ZENG Q,YAN F M,et al.Effects of transgenic plants on soil microorganisms[J].Plant and Soil,2005,271(1/2):1-13.
[8] YANG Y,SHI W Y,WAN M X,et al.Diversity of bacterial communities in acid mine drainage from the Shen-bu copper mine,Gansu province,China[J].Electron Journal of Biotechnology,2008,11(1):107-118.
[9] MWIRICHIA R,COUSIN S,MUIGAI A W,et al.Archaeal diversity in the haloalkaline Lake Elmenteita in Kenya[J].Current Microbiology,2010,60(1):47-52.  
[10] ROBERFROID M B,BORNET F,BOULEY C,et al.Colonic microflora:nutrition and health.Summary and conclusions of an International Life Sciences Institute (ILSI)[Europe] workshop held in Barcelona,Spain[J].Nutrition Reviews,1995,53(5):127-130.
[11] CUMMINGS J H,BEATTY E R,KINGMAN S M,et al.Digestion and physiological properties of resistant starch in the human large bowel[J].British Journal of Nutrition,1996,75(5):733-747.  
[12] BÄCKHED F,DING H,WANG T,et al.The gut microbiota as an environmental factor that regulates fat storage[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(44):15718-15723.  
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