Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (6): 81-92.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1102

Previous Articles     Next Articles

Effects of Analysis Methods on the Analyzed Results of 16S rRNA Gene Amplicon Sequencing in Bacterial Communities

ZHONG Hui1,2(), LIU Ya-jun1,2, WANG Bin-hua1,2, HE Meng-jie1,2, WU Lan1,2()   

  1. 1. College of Life Sciences,Nanchang University,Nanchang 330031
    2. Ministry of Education,Key Laboratory of Environment and Resource Utilization of Poyang Lake,Nanchang University,Nanchang 330031
  • Received:2021-08-27 Online:2022-06-26 Published:2022-07-11
  • Contact: WU Lan E-mail:udaio@qq.com;ncusk724@hotmail.com

Abstract:

Bacterial 16S rRNA gene amplicon sequencing is one of the most widely used method in environmental microbiology. However,there are many ways to determine the minimum taxonomic unit of the sequencing reads,and its effect on the downstream analysis results of microbial diversity needs to be further explored. This study extracted DNA from 5 groups of environmental samples(forest soil,farmland soil,wetland soil,lake sediment,and lake water)for 16S rRNA gene amplicon sequencing,and based on different taxonomic resolutions,5 types of minimum taxonomic unit division method(Sequence clustering OTUs at 97%,98%,99%,100% similarity,and the DADA 2 algorithm to calculate ASVs)were used to compare and analyze the influences of minimum taxonomic unit division methods on microbial community diversity,composition and its correlation with environmental factors. The results showed that higher community α diversity(Chao1 and Shannon)and β diversity(P < 0.05)were obtained by increasing the taxonomic resolution. Compared with the OTU,which was clustering by sequence similarity,the ASV method reduced Chao1 and PD indices to some extent. For community composition,the division method of the taxonomic unit mainly affected the proportion of some low-abundance genera(< 0.2%),but had a smaller effect on the composition of higher taxonomy level(phylum level). In addition,the results of redundant analysis demonstrated that increasing the level of taxonomic resolution would allow the environmental factors to have a higher interpretation of the microbial community,and a concurrently it also affected the order of interpretation degree of community composition by environmental factors. In sum,this study clarified the effect of different division methods of the minimum taxonomy unit on the diversity,composition and association with environmental factors,which provides theoretical guidance for the subsequent study of environmental microbiomics.

Key words: environmental microbiomics, high-throughput sequencing, 16S rRNA gene, minimum taxonomy unit, bacterial diversity