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.