变性梯度凝胶电泳法初步解析茯砖茶渥堆发酵过程中细菌群落结构  

Preliminary Analysis of Bacterial Community Structure during Pile-Fermentation Process of Fuzhuan Brick Tea by Denaturing Gradient Gel Electrophoresis(DGGE)

作　　者：刘石泉[1,2] 胡治远[1] 赵运林[1] 

机构地区：[1]湖南城市学院化学与环境工程学院,湖南益阳413000 [2]湖南农业大学生物科学与技术学院,湖南长沙410128

出　　处：《食品科学》2014年第15期172-177,共6页Food Science

基　　金：湖南省自然科学基金项目(13JJ6074);湖南省高校产学研合作示范基地产业化项目(13CY025;11CY003;11CY004);湖南省科学技术厅科技计划国际合作项目(2012WK2013);湖南省高校科技创新团队支持计划资助项目(2014);益阳市科学技术局科技计划项目(2013YK1323)

摘　　要：为解析茯砖茶渥堆发酵过程中细菌群落结构和种类,对渥堆过程中不同时间段细菌16S rDNA的V3可变区进行扩增,对细菌变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)图谱中条带进行克隆、测序和序列比对。结果表明:黑毛茶在渥堆过程中以渥堆24 h为分界点,前后各自细菌群落结构相似,但前后的差异较大;从16S rDNA的V3可变区比对结果证明黑毛茶渥堆过程中有诺卡氏菌属、新鞘脂菌属、短波单胞菌属、韦龙氏假单胞菌属、突那梭菌属、克雷伯氏菌属、乳杆菌属及不可培养的ε-变形菌、腐败螺旋菌属、黏球菌属、根瘤菌属和未知分类地位的不可培养细菌6种。采用DGGE指纹图谱能更全面、更真实地反映黑毛茶渥堆发酵过程中细菌群落的结构和多样性变化。In order to explore the bacterial community structure and species during the pile-fermentation process of Fuzhuan Brick Tea, the V3 variable region of bacterial 16 S rDNA was amplified at different fermentation times and analyzed by denaturing gradient gel electrophoresis(DGGE), and the DGGE bands were cloned, sequenced and aligned. The results showed that 24 hours of pile-fermentation was the critical point. A similar bacterial community structure was observed before and after the point, respectively, while a significant difference existed between the two stages. The sequence alignment of 16 S rDNA V3 variable region showed that Millisia brevis, Novosphingobium sp., Brevundimonas aurantiaca, Pseudomonas veronii, Clostridium ultunense, Klebsiella pneumonia, Lactobacillus plantarum, unculturable strains including Epsilon proteobacterium, Saprospiraceae bacterium, Myxococcales bacterium and Rhizobiales bacterium, and six species of unculturable bacteria with unknown taxonomic status were detected. Therefore, pile fermentation of the black tea involved a large number of bacteria, and the bacterial community during the process was relatively stable. In conclusion, DGGE fingerprint can provide a comprehensive and true reflection of changes in the bacterial community structure and diversity of Fuzhuan brick tea during the fermentation process.

关 键 词：茯砖茶 渥堆发酵 变性梯度凝胶电泳 细菌群落结构 

分 类 号：TS272[农业科学—茶叶生产加工]