王艳丽,张会敏,孟雅静,周庆伍,梁金辉,袁志强,刘国英,何宏魁,李安军.基于16S rDNA V4与V3-V4可变区高通量测序对浓香型白酒窖泥菌群组成的比较分析[J].,2020,36(6):147-154.
基于16S rDNA V4与V3-V4可变区高通量测序对浓香型白酒窖泥菌群组成的比较分析
Comparative Analysis of Strong-flavor Baijiu Pit-mud Microbial Community Using High-throughput Sequencing Based on 16S rDNA V4 and V3-V4 Variable Regions
投稿时间:2019-11-20  
DOI:10.13982/j.mfst.1673-9078.2020.6.1130
中文关键词:  Illumina高通量测序  窖泥菌群  V4区  V3~V4区
英文关键词:illumina high-throughput sequencing  Pit-mud microbial community  V4 variable  V3-V4 variable
作者简介:王艳丽(1989-),女,硕士,研究方向:发酵微生物及其应用 通讯作者:李安军(1970-),男,高级工程师,研究方向:酿酒生产工艺技术、科研开发、质量管理工作
基金项目:
作者单位
王艳丽 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
张会敏 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
孟雅静 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
周庆伍 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
梁金辉 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
袁志强 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
刘国英 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
何宏魁 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
李安军 (安徽省固态发酵工程技术研究中心,安徽亳州 236820) 
AuthorInstitution
WANG Yan-li (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
ZHANG Hui-min (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
MENG Ya-jing (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
ZHOU Qing-wu (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
LIANG Jin-hui (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
YUAN Zhi-qiang (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
LIU Guo-ying (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
HE Hong-kui (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
LI An-jun (The Center for Solid-state Fermentation Engineering of Anhui Province, Bozhou, 236820) 
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中文摘要:
      本研究分别对新老窖泥样本的16S rDNA V4和V3-V4区进行Illumina高通量测序,比较了测序的群落多样性参数、NMDS聚类与优势门/属的相对丰度。V4区的OTUs数目(264~639)显著低于V3-V4区(459~1123);V4区的Shannon指数在新、老池壁泥中(2.47、5.02)显著低于V3-V4区(3.33、6.46),而在新(4.98vs5.98)、老(5.64vs5.39)池底泥样本中无显著差异(p?0.05);V4区的Chao1指数在新池底泥(2455vs1358)、新池壁泥(1326vs504)和老池底泥(3475vs1260)中显著高于V3~V4区,在老池壁泥中无显著差异(1915vs1312)(p?0.05)。两个可变区的测序对NMDS聚类规律无影响。热图聚类结果显示,两个可变区的优势门/属的相对丰度很接近,V4区在属水平上更容易区分老窖池的池底泥。与V3-V4区相比,V4区更利于区分池壁泥与池底泥,且可以检测出甲烷菌的含量,更有利于窖泥菌群组成分析。本研究为16S rDNA高通量测序中V3-V4区与V4区的选择提供一定参考。
英文摘要:
      In this study, Illumina high-throughput sequencing of 16S rDNA V4/V3-V4 variable regions of old and young pit-mud (PM) samples were performed. The diversity indices, NMDS clustering results, and the relative abundance of dominant phyla/genera of two variable regions were compared. The number of OTUs in V4 (264~639) were significantly lower than that of V3-V4 (459~1123). The Shannon indices of V4 in the young (2.47) and old (5.02) Wall-PM (WPM) samples were significantly lower than those of V3-V4 (3.33 and 6.46, respectively). Insignificant difference (p?0.05) was found between the young (4.98vs5.98) and old (5.64vs5.39) bottom-PM (BPM) samples. The Chao1 indices of V4 in the young BPM (2455vs1358), young WPM (1326vs504) and old BPM (3475vs1260) were significantly higher than those of V3-V4, with insignificant difference (p?0.05) in the old WPM samples (1915vs1312). The sequencing of the two variable regions had no effect on the NMDS clustering results. The heat-map clustering results showed that the relative abundance of the dominant phyla/genera for two variable regions were pretty similar, while it was easier for theV4 variable region to distinguish old BPM samples at the genus level. Compare with the V3-V4 region, the V4 variable region was more conducive to distinguishing between WPM and BPM and detecting the content of methanogen, which allows efficient analysis of the PM microbial composition. This study provided a good reference for the selection of V3-V4 and V4 regions during high-throughput sequencing of 16S rDNA.
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