酒鬼酒成熟糖化料细菌的分离和系统发育多样性分析  

Isolation and Phylogenetic Diversity Analysis of Bacteria from the Mature Saccharified Grains of Jiuguijiu Liquor

作　　者：赫立娟 陈锦华 邓丽颖 刘祝祥[1] 贺建武[1] 陈义光[1] HE Lijuan;CHEN Jinhua;DENG Liying;LIU Zhuxiang;HE Jianwu;CHEN Yiguang(College of Biology and Environmental Sciences,Jishou University,Jishou,Hunan 416000;Dachang Tiangezhuang School,Langfang,Hebei 065300,China)

机构地区：[1]吉首大学生物资源与环境科学学院,湖南吉首416000 [2]大厂回族自治县田各庄学校,河北廊坊065300

出　　处：《酿酒科技》2021年第7期22-28,共7页Liquor-Making Science & Technology

基　　金：国家自然科学基金(31460004,31660003);酒鬼酒股份有限公司委托项目(JGKJ 2012-JD03)。

摘　　要：采集酒鬼酒成熟糖化料样品,采用纯培养法以3种基础分离培养基分离样品中的细菌(含放线菌),采用16S rRNA基因序列分析法对分离菌株进行发育多样性分析。用营养琼脂(nutrient agar)、LA(Luria-Bertani agar)和海洋琼脂(marine agar)培养基进行分离,得到82个细菌菌株。根据菌落形态、细胞形态和革兰氏染色实验结果,去除部分冗余,最终得到47个代表性菌株。系统发育分析结果表明,所得47个代表性菌株分属于细菌域(Bacteria)的5个大的系统发育类群/门(phylogenetic group/phylum),近半数菌株(23株,48.9%)属于厚壁菌门,其中优势菌群为芽孢杆菌科(Bacillaceae)的芽孢杆菌属(Bacillus),共有11个分离菌株(23.4%)。这47个代表性菌株可以归为31个不同物种,且大多数与它们的系统发育关系最密切的已知物种的典型菌株之间存在一定的遗传差异,16S r RNA基因序列相似性处于96.52%~99.73%之间。其中有2个菌株(JSM 2032011,JSM 2032014)与其系统发育关系最密切的已知物种的典型菌株之间的16S rRNA基因序列相似性较低,可能代表潜在的新分类单元(potential new taxa)。菌株JSM 2032011与假单胞菌属(Pseudomonas)的Pseudomonas hunanensis LVT的系统发育关系最密切,但它们之间的序列相似性较低(97.54%);菌株JSM 2032014与赖氨酸芽胞杆菌属(Lysinibacillus)的Lysinibacillus jejuensis N2-5T的系统发育关系最密切,它们之间的序列相似性只有96.52%。而且这2个菌株在系统发育树上在各自的簇群中形成了独立的进化分支。因此,菌株JSM 2032011和JSM2032014可能分别代表了假单胞菌属和赖氨酸芽胞杆菌属的潜在新种(potential new species)。以上结果表明,酒鬼酒成熟糖化料分离的细菌具有较高的类群多样性和物种多样性,且蕴含着一些新的微生物类群/物种。In this study,the mature saccharified grain samples of Jiuguijiu Liquor were collected,and 3 media(nutrient agar,LuriaBertani agar and marine agar)were used to isolate bacteria strains from the samples by pure cultivation method.The phylogenetic diversity of the isolated strains was studied based on 16 S r RNA gene sequences.A total of 82 bacteria strains were obtained.Based on colonial morphology,cell morphology,and Gram staining tests,47 representative strains were selected for further study.The results of phylogenetic analysis showed that,the 47 strains belonged to 5 phylogenetic groups/phyla;23 strains(48.9%)belonged to Firmicutes,among which 11 strains(23.4%)were Bacillus;the 47 strains belonged to 31 different species,and the similarity between their 16 S rRNA sequences and that of the type strains were in the range of 96.52%to 99.73%.Strain JSM 2032011 was most closely related to Pseudomonas hunanensis LV^(T),while the gene sequence similarity between them was merely 97.54%.Strain JSM 2032014 was most closely related to Lysinibacillus jejuensis N2-5^(T),while the gene sequence similarity between them was merely 96.52%.In addition,strain JSM 2032011 and strain JSM 2032014 respectively formed distinct branches in the phylogenetic tree,which indicated that strain JSM 2032011 and strain JSM 2032014 might represent potential new species of Pseudomonas and Lysinibacillus,respectively.In conclusion,the bacteria strains isolated from the mature saccharified grains of Jiugui Liquor had high diversity,with potential new taxa and species.

关 键 词：酒鬼酒 成熟糖化料 细菌 纯培养 系统发育分析 

分 类 号：TS262.3[轻工技术与工程—发酵工程] TS261.1[轻工技术与工程—食品科学与工程]