基于高通量测序的表层水体纤毛虫多样性评估方法比较与优化OA北大核心CSTPCD

High-throughput amplicon sequencing has been widely employed for studying microbial diversity.Dif-ferent sequencing methods and data analysis processes influence the analysis results of microbial biodiversity and community structure.Therefore,this study aimed to investigate the effects of different methods,the key steps in-volved in data analysis of the chimera removal and rare taxa,and the merging of similar taxonomic units.Results revealed that the main species detected by DNA sequencing,RNA sequencing,and morphological methods were similar.Moreover,fewer species were detected by RNA sequencing than by DNA sequencing;however,the differ-ence was not significant.Community structural analysis showed a similar community structure based on the opera-tional taxonomic unit(OTU)clustering on a similarity of 97%,98%,99%,and 100%.Both methods of the single-nucleotide variation and cluster based on sequence similarity were able to detect a similar relative abundance of different taxa to that by the morphological method.The species richness detected by OTU clustering under 99%similarity was similar to that detected by morphological results.Furthermore,chimera removal and rare taxa(re-moval thresholds:0.05%for DNA sequencing and 0.07%for RNA sequencing)can improve the issue of inflated molecular diversity.Thus,this study provides a scientific analysis process to investigate ciliate molecular diversity and provides guidelines for future research on eukaryotic microbial diversity.