PCR-DGGE法分析泡菜中酵母菌微生物的多样性  

Analysis of Microbial Diversity of Yeast in Pickles by PCR-DGGE Method

作　　者：燕平梅[1,2] 李安萍 YAN Ping-mei;LI An-ping(Department of Biology,Taiyuan Normal University,Taiyuan 030619,China;Green Industry Technology Innovation Strategic Alliance of Soil Disinfestation and Activation,Taiyuan 030619,China)

机构地区：[1]太原师范学院生物系,太原030619 [2]土壤消毒活化绿色产业技术创新战略联盟,太原030619

出　　处：《中国调味品》2020年第12期23-26,共4页China Condiment

基　　金：国家自然基金项目(31171743);山西省重点研发项目(201603D221008-1)。

摘　　要：目的:采用非培养法直接分析市场上3种不同市售泡菜中酵母菌的多样性。方法:以市场上的泡菜汁中总基因DNA为模板,采用酵母菌引物(NL1-GC/LS-2)进行PCR扩增,通过变性梯度凝胶电泳(DGGE)分离不同泡菜中提取的总微生物混合物真菌26S rDNA,再利用Quantity One软件分析DGGE图谱,得出其多样性指数、均匀度指数和丰富度指数,从而分析鉴定这3种泡菜中酵母菌的多样性;通过UPGAME算法对3种泡菜的酵母菌群落结构进行相似性聚类分析;采用MEGA 6.06软件构建系统发育树。结果:3种泡菜中共有3种不同的酵母菌,分别为Pichia fermentans(毕赤酵母菌)、Candida tropicalis(热带假丝酵母菌)、Saturnispora mendoncae。散称泡菜(P1)中有1种酵母菌,白菜泡菜(P2)中有3种酵母菌,泡酸菜(P3)中有2种酵母菌;其中Saturnispora mendoncae是3种泡菜中共有的酵母菌,Candida tropicalis(热带假丝酵母菌)是P2中特有的1种酵母菌,Pichia fermentans(毕赤酵母菌)是P2和P3共有的。多样性指数P2最大,均匀度指数P1最大。Objective:The non-culture method is used to directly analyze the diversity of yeast in three kinds of pickles on the market.Method:Using the total gene DNA in pickle juice on the market as the template,PCR amplification is performed using yeast primer(NL1-GC/LS-2).The total microbial mixture of fungi 26S rDNA extracted from different pickles is isolated by denaturing gradient gel electrophoresis(DGGE).Quantity One software is used to analyze the DGGE map,and the diversity indexes,evenness indexes and richness indexes are obtained,so as to analyze and identify the diversity of yeast in these three pickles.UPGAME algorithm is used for the similarity and clustering analysis of the community structure of yeast in the three kinds of pickles.The phylogenetic trees are constructed with MEGA 6.06 software.Results:There are three different kinds of yeast such as Pichia fermentans,Candida tropicalis and Saturnispora mendoncae in the three kinds of pickles.There's one kind of yeast in pickles in bulk(P1),three kinds of yeast in cabbage pickles(P2),and two kinds of yeast in Chinese sauerkraut(P3).Among them,Saturnispora mendoncae is a common yeast among the three kinds of pickles,Candida tropicalis is a special yeast in P2,and Pichia fermentans is a common yeast in P2 and P3.The diversity indexes of P2 are the largest and the evenness indexes of P1 are the largest.

关 键 词：泡菜 酵母菌多样性 PCR DGGE 

分 类 号：TS255.54[轻工技术与工程—农产品加工及贮藏工程]