高通量测序技术解析五粮液窖泥原核微生物群落结构

摘要

In order to well-understand the specific brewing microbial community of Wuliangye pit mud and its function.The 16S ribosomal DNA V4 sequences of prokaryotic community in Wuliangye pit muds from different positions were investigated by Illumina NextSeq 500 next-generation sequencing (NGS) platform for the first time.Numbers of DNA sequences and operation taxonomy units (OTUs) were summarized by optimized QIIME program,and the abundance and diversity indices of prokaryotic microbial community were calculated.A total of 7 555 164 high quality sequences were obtained and under the same sequencing depth,bacteria (92.3％) and archea (7.6％) were determined as main microbial communities.22 phyla,52 classes,89 orders,168 family,330 genera were identified by optimized alignment method.Fimicutes was the predominant bacteria in all pit muds with the abundance of 85.6％,Clostridium,Lactobacillus,Coprococcus,Caldicoprobacter,and Soehngenia were dominant genus.Furthermore,according to the phylogenetic analysis of dominant OTUs (＞ 1％),the dominant genus in pit mud included Lactobacillus,Caldicoprobacter,Caloramater,Clostridium,Caloribacterium,Garciella,Eubacterium,Syntrophomonas,Sedimentbacter,Sporanaerobacter,Tissierella,Methanosarcina,Methanobacterium,Methanobrevibacter,and Methanoculleus.Wuliangye pit mud contained the specific prokaryotic community in comparison with other Strong-aroma liquors,and it consisted of complex Firmicutes,high percentage of hexanoic acid-production community and methane community.The specific prokaryotic community can promote the formation of hexanoic acid.%为清晰认识五粮液特殊窖泥微生态群落结构及功能,利用Illumina NextSeq 500高通量深度测序平台首次分析了五粮液不同窖泥原核微生物群落16S rDNA V4高变区序列;优化了QIIME程序并整理和统计样品序列数目和操作分类单元(OTUs)的数量,分析了不同窖泥中原核微生物群落的丰度、多样性;利用系统发育分析确定了优势菌群的系统发育地位.该研究结果表明,从五粮液窖泥中共获得7 555 164条高质量目标片段.在相同的测序深度条件下(300 000条),五粮液窖泥原核微生物主要由细菌和古菌构成,共检出22个门,52个纲,89个目,168个科,330个属原核微生物,细菌占92.3％,古菌占7.6％.厚壁菌门(Fimicutes)是所有样品的绝对优势菌群(85.6％),共检出74个属,Clostridium、Lactobacillus,Coprococcus,Caldicoprobacter,Soehngenia是丰度最高的细菌属.结合系统发育分析确定窖泥中优势菌群为Lactobacillus,Caldicoprobacter,Caloramater,Clostridium,Caloribacterium,Garciella,Eubacterium,Syntrophomonas,Sedimentbacter,Sporanaerobacter,Tissierella,Methanosarcina,Methanobacterium,Methanobrevibacter,Methanoculleus.五粮液窖泥中蕴藏了显著区别于其他浓香型白酒窖泥的原核微生物群落,尤其是群系复杂的厚壁菌门细菌,较高丰度的产己酸菌群和促进己酸生成的甲烷菌群.