基于不同测序技术的微生物群落结构比较  

Comparison of microbial community structure based on different sequencing techniques

作　　者：黄纷 岳正波[1] 李玉龙[1] 谭娟 王进[1] HUANG Fen;YUE Zhengbo;LI Yulong;TAN Juan;WANG Jin(School of Resources and Environmental Engineering,Hefei University of Technology,Hefei 230009,China)

机构地区：[1]合肥工业大学资源与环境工程学院,安徽合肥230009

出　　处：《合肥工业大学学报（自然科学版）》2020年第12期1676-1683,共8页Journal of Hefei University of Technology：Natural Science

基　　金：国家自然科学基金面上资助项目(41572326,41772361)。

摘　　要：文章分别采用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)、克隆文库(clone library,CL)及16S rRNA高通量测序(high-throughput sequencing,HS)技术,分析厌氧-缺氧-好氧(anaerobic-anoxic-oxic,A 2/O)反应器稳定运行期间各个反应器内微生物群落结构多样性,评价3种测序技术在具体应用中的特色及其局限性。结果表明:对于厌氧反应器古菌分析,HS比DGGE和CL分别多检测出50.00%、83.00%,而在细菌分析中,HS比DGGE和CL分别多检测出21.42%、50.00%;对于缺氧反应器和好氧反应器中细菌的群落结构分析,3种方法测得的微生物种类差别不大。CL操作简单,适用于样品量少的情况;DGGE能直观地反映微生物群落的变化情况,但1次最多分析32个样品;而在分类水平低时,CL和DGGE会低估样品中微生物物种组成,高估相对丰度。In this paper,denaturing gradient gel electrophoresis(DGGE),clone library(CL)and 16S rRNA high-throughput sequencing(HS)technology were used to analyze the diversity of microbial community structure in each reactor during the stable operation of anaerobic-anoxic-oxic(A 2/O)reactor and the characteristics and limitations of three different sequencing technologies in specific applications were evaluated.In the process of archaea analysis in anaerobic reactor,by comparing with DGGE and CL,results showed that HS detected more than 50.00%and 83.00%of microbial species,respectively,while in the bacteria analysis,it detected more than 21.42%and 50.00%of microbial species,respectively.Furthermore,the three methods showed little difference in microbial species for the community structure analysis of bacteria and archaea in anoxic and aerobic reactors.The CL is easy to operate and suitable for a small number of samples.The DGGE analysis of up to 32 samples can directly reflect changes of microbial communities,but when the classification level is low,the CL and DGGE will underestimate the microbial species composition and overestimate relative abundance in the sample.

关 键 词：高通量测序(HS) 变性梯度凝胶电泳(DGGE) 克隆文库(CL) 厌氧-缺氧-好氧(A^2/O)反应器 微生物群落 

分 类 号：X172[环境科学与工程—环境科学]