采用免培养(culture independent)技术直接从浓香型白酒大曲中提取细菌微生物基因组DNA,利用细菌16S rDNA通用引物扩增大曲混合微生物的16S rDNA,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大曲中细菌的16S rDNA基因全序列,通过与基因数据库中相似菌群序列同源性的比较,构建系统发育树。结果显示,成熟的大曲中细菌共分为Lactobacillus,Pantoea,Enterobacter,Klebsiella,Leuconostoc,Erwinias,Pseudomonas,Bacillus licheniformis几大类群,表现出高度的细菌多样性。

Culture independent method was used to study the diversity of Daqu bacteria.The techniques including culture independent approach,PCR technique and 16S rDNA sequence homology analysis etc.The total DNA directly extracted from Daqu was used as PCR template.Random clones,containing almost full size 16S rDNA sequences(about 1.5 kb long),were sequenced and subjected to an on line similarity search.According to the sequences alignment and through the analysis of its comparability,the bacteria of Daqu were consisted of Lactobacillus,Pantoea,Enterobacter,Klebsiella,Leuconostoc,Erwinias,Pseudomonas and Bacillus licheniformis,which indicated their bacterial diversity.