16S rDNA克隆文库法与高通量测序法在浓香型大曲微生物群落结构分析中的对比研究  

16S rDNA Clone Library vs.High-throughput Sequencing Method in the Analysis of the Microbial Communities in Nongxiang Daqu

作　　者：陈玲[1] 袁玉菊[1] 曾丽云[1] 廖作敏 陈伊童 吴正云[1] 张文学[1,2] 

机构地区：[1]四川大学轻纺与食品学院,四川成都610065 [2]四川大学锦江学院白酒学院,四川眉山620860

出　　处：《酿酒科技》2015年第12期33-36,40,共5页Liquor-Making Science & Technology

基　　金：四川大学德阳校市科技合作专项资金项目(014CDD-S03-SCU);国家自然科学基金(31071585)

摘　　要：针对浓香型白酒大曲样本,以16S r RNA基因为目的片段,分别采用16S r DNA克隆文库法和高通量测序法分析大曲中细菌微生物群落的组成,并通过丰度和多样性分析,比较了两种方法在研究大曲样品细菌多样性方面的适用性。结果表明,在门、纲、目、科和属的分类水平上,克隆文库方法检测大曲样本微生物得到4个门,4个纲,5个目,4个科,6个属;高通量测序得到13个门,22个纲,33个目,61个科,133个属。在门的水平上,克隆文库与高通量测序检测出优势类群的总数量与总丰度分别为3个(99.32%)和4个(98.61%),共有优势类群及其丰度分别为Firmicutes(88.88%和79.32%)、Proteobacteria(7.8%和15.04%)、Actinobacteria(2.72%和1.77%)。重复样本分析,得出的结果相似。克隆文库法与高通量测序法在反映样本微生物群落规模上差异较大,而在反应大曲样本中主要微生物的物种组成及数量比例上结果相近,特别是样本中优势微生物类群的结果基本相同。两种方法各具优势。16S r DNA clone library and high-throughput sequencing method were used respectively to analyze the microbial communities in Nongxiang Daqu（Nongxiang Daqu used as the research object and 16 S r RNA gene as the target fragments）. The suitability of the two methods for the research on bacterial diversity in Daqu was compared through richness diversity analysis. The results revealed that, at the taxonomic levels of microbes including phylum, class, order, family, and genus, clone library method could detect 4 phylum,4 class,5 order, 4 family and6 genus in total, while high-throughput sequencing method could detect 13 phyla, 22 class, 33 order, 61 family and 133 genus; at phylum taxonomic level, there were 3 and 4 dominant phylum detected by clone library and high-throughput sequencing respectively, and the total richness were 99.32 % and 98.61 % respectively, and phylum such as Firmicutes, Proteobacteria, and Actinobacteria were the dominant microbes in both 16 S r DNA clone library and high- throughput sequencing, and their richness were 88.88 % vs. 79.32, 7.8 % vs. 15.04, and 2.72 % vs.1.77 % respectively. The repeated samples analysis got the same results. In conclusion, there was significant difference in reflecting microbial community size in Daqu between 16 S r DNA clone library and high-throughput sequencing method. However, both methods got close detection results in the main microbial community composition quantity ratio especially in the dominant microbial community of Daqu. Both methods had their advantages.

关 键 词：16S r DNA克隆文库 高通量测序 大曲 微生物群落 白酒 

分 类 号：TS261.1[轻工技术与工程—发酵工程] Q93-3[轻工技术与工程—食品科学与工程]