传统分离培养结合DGGE法检测榨菜腌制过程的细菌多样性.docVIP

传统分离培养结合DGGE法检测榨菜腌制过程的细菌多样性 李正国1* 付晓红1 邓 伟1 杨迎伍1 崔英双2 赵 平2 (1. 重庆大学生物工程学院基因工程研究中心 重庆市高校功能基因与调控新技术重点实验室 重庆 400044)(2. 重庆涪陵榨菜(集团)有限公司 重庆 408000) 摘 要: 采用传统分离培养和基于16S rRNA 作为分子标记的变性梯度凝胶电泳(Denaturing gradient gel electrophoresis, DGGE)的方法, 分析榨菜腌制过程中不同时期的可培养细菌数量、多样性及其群落结构。结果表明, 用传统分离与分子鉴定方法获得7个属的细菌类群, 其中乳杆菌属(Acidobacterium)是优势菌群, 明串珠菌属(Leuconostoc)是次优势菌群。对通过DGGE方法得到的11条16S rRNA优势条带序列进行了比对, 结果表明明串珠菌属(Leuconostoc)的丰度最高, 是腌制过程中主要的优势菌群, 乳球菌(Lactococcus lactis)是次优势菌。传统分离法与DGGE法结合能够更有效的分析榨菜腌制过程中微生物的多样性及其动态变化, 获得更全面的微生物多样性信息。 关键词: 榨菜(Brassica juncea coss var. tsatsai), 16S rRNA, 传统分离培养, 变性梯度凝胶电泳(DGGE), 群落多样性 Analysis of Bacterial Diversity During the Processing of Brassica juncea coss var. tsatsai by the Culture-independent and Denaturing Gradient Gel Electrophoresis (DGGE) Methods LI Zheng-Guo1* FU Xiao-Hong1 DENG Wei 1 YANG Ying-Wu1CUI Ying-Shuang2 ZHAO Ping2 (1. Genetic Engineering Research Center, Bio-Engineering College, Chongqing University, Key Lab of Functional Gene andNew Regulation Technologies under Chongqing Municipal Education Commission, Chongqing 400044, China)(2. Chongqing Fuling Zhacai Group Ltd., Chongqing 408000, China) Abstract: Both culture-dependent and culture-independent methods, denaturing gradient gel electrophoresis (DGGE) based on the sequence of 16S rRNA V3 region gene, were used to examine the total bacteria counts, bacterial diversity and community structure of different periods Brassica juncea saline water samples. The results showed that 7 genera of bacteria were identified from 19 isolated bacterial populations by the traditional isolation method, the dominant bacteria in intestine belonged to Acidobacterium. By 16S rRNA V3 region gene DGGE method, 11 distinct bands were obtained from 16S rDNA amplifications. The profile of DGGE fingerprints of different periods saline water revealed that the most dominant bacteria group was Leuconostoc in the process of the fermentation. This suggested that a combination of molecular an