西藏地区传统发酵乳宏基因组DNA的提取及微生物群落多样性分析  

Metagenomic DNA Extraction and Analysis of Microbial Diversities of Traditional Fermented Dairy Products in Tibet

作　　者：张春林[1] 刘文俊[1] 张彦斌[1] 孙志宏[1] 张和平[1] 孙天松[1] 

机构地区：[1]内蒙古农业大学乳品生物技术与工程教育部重点实验室

出　　处：《食品与发酵工业》2010年第4期15-20,共6页Food and Fermentation Industries

基　　金：国家自然基金项目(No.30660135;30800861);现代农业产业技术体系项目;内蒙古自然科学基金项目(2009MS0409)

摘　　要：采用CTAB-SDS-冻融、液氮研磨、玻璃珠吸附3种方法提取来自西藏地区的传统发酵乳样品中微生物宏基因组DNA。结果表明,采用经过改良的CTAB-SDS-冻融法提取效果较佳,可直接用于后续的微生物多样性分析。分别采用16SrRNA-RFLP和ITS-PCR分析发酵乳中细菌和真菌的微生物多样性,结果显示,来自西藏当雄龙仁乡的2个样品中微生物的多样性较为丰富,并且优势菌群较为相近,而来自西藏当雄纳木错的样品微生物多样性丰度较低,其优势菌群与前2个样品有较大的差异性。此结果同时表明了运用细菌16SrRNA-RFLP和真菌ITS-PCR相结合的方法能够全面快速地比较分析发酵乳中的微生物多样性。Three methods, CTAB-SDS-frozen-thawed, liquid nitrogen trituration and glass bead adsorptive process, were used to extract the metagenomic DNA of microbes in traditional fermented dairy products from Tibet, China. The results showed that modified CTAB-SDS- frozen-thawed can give a best extracting products, and the products can be used for further microbial diversity analysis. This research also analyzed the microbioal diversities of the samples using 16S rRNA-RFLP and ITS-PCR methods, the results showed that the samples of Longren township in Dangxiong province of Tibet expressed much more plenty microbial diversity, the dominant microbiology in these two samples also showed a high similarity. However, the microbiology in the samples of Namucuo township in Dangxiong province of Tibet showed a low abundance, and the dominant microbiology also expressed high difference with the samples of Longren township. The results illuminated that combined 16S rRNA-RFLP and ITS-PCR are powerful tool for estimating the community diversity of bacteria and true fungus in fermented dairy sample.

关 键 词：传统发酵乳 宏基因组DNA 微生物群落多样性 PCR—RFLP ITS—PCR 

分 类 号：TS252.1[轻工技术与工程—农产品加工及贮藏工程]