酱香型窖泥古菌群落结构的研究

Research on Archaea Community Structure in the Pit Mud of Maotai-flavor Liquor

采用ARDRA免培养手段研究酱香型窖泥中的古菌群落结构,通过对窖泥总DNA的提取,扩增古菌16SrDNA序列,构建古菌16S rDNA文库。随机挑选39个阳性克隆子,通过HhaI限制性内切酶酶切,选择酶切图谱不同的25个克隆子用于后续测序,测序结果与NCBI数据库比对分析,获得其分类信息。克隆文库分析结果表明,酱香型窖泥中古菌主要分布于广古菌门中的甲烷袋状菌属(Methanoculleus)、甲烷八叠球菌属(Methanosarcina)、甲烷鬃毛菌(Methanosaeta)和甲烷杆菌属(Methanobacterium),分别占44%、41%、3%、9%。

The purpose of this research is to study the structure of archaea community in the pit mud of Maotai-flavor liquor through a culture-in- dependent method named ARDRA. The clone species information was collected through extracting total DNA of pit mud, amplifying 16S rDNA sequences ofarchaea, constructing archaea 16S rDNA library and sequencing. 39 positive clones were selected randomly, and then digested by re- strict enzyme HhaI.25 different clones with different restrict enzyme diagram were selected for subsequent sequencing. Then the classification in- formation of different clone was analyzed by comparing its sequencing results with NCBI database. The analytical results showed that archaea community in pit mud of Maotai-flavor liquor was mainly comprised of MethanocuUeus（44 %）, Methanosarcina （41%）, Methanosaeta （3 %）, and Methanobacterium（9 %）.