采用免培养(culture independent)技术直接从浓香型白酒窖泥中提取细菌微生物混合基因组DNA(也叫元基因组DNA),利用细菌16S rDNA通用引物扩增窖泥细菌的序列,根据16S rDNA序列对细菌多样性进行初步分析。采用PCR扩增技术、分子克隆技术以及序列同源性分析等方法测定细菌的16S rDNA,通过与基因数据库中相似菌群序列同源性的比较,得到样品菌种多样性,分别构建系统发育树。结果显示,细菌含有几大类群,表现出高度的细菌多样性。共分为Uncultured bacterium、Clostridium、Lactobacillus、Eubacterium和Syntroph-omonas五个细菌分类。

Culture independent method was used to study the diversity of Pit Mud bacteria.The techniques included culture independent approach,PCR technique and 16S rDNA sequence homology analysis etc.The total DNA directly extracted from Pit Mud was used as PCR template.Random clones containing almost full size 16S rDNA sequences(about 1.5 kb) were sequenced and subjected to an on line similarity search.According to the sequences alignment and homology analysis,the bacteria of Pit Mud were consisted of Uncultured bacterium,Clostridium,Lactobacillus,Eubacterium,and Syntrophomonas.