Improving the microbial community reconstruction at the genus level by multiple 16S rRNA regions

• A method to measure the proper region for microbial community reconstruction.
• The preferential genus-specific amplicons are not always located in HVRs.
• Low-abundance genera should may be ignored by “universal” primers.
• Using multiple regions can improve the microbial community reconstruction.

Background16S rRNA genes have been widely used for phylogenetic reconstruction and the quantification of microbial diversity through the application of next-generation sequencing technology. However, long-read sequencing is still costly, while short-read sequencing carries less information for complex microbial community profiling; therefore, the applications of high throughput sequencing platforms still remain challenging in microbial community reconstruction analysis.ResultsHere, we developed a method to investigate the profile of aligned 16S rRNA gene sequences and to measure the proper region for microbial community reconstruction, as a step in creating a more efficient way to detect microorganism at the genus level. Finally, we found that each genus has its own preferential genus-specific amplicons for a genus assignment, which are not always located in hyper variable regions (HVRs). It was also noted that the rare genera should contribute less than dominant ones to the common profile of the aligned 16S rRNA sequences and have lower affinity to the common universal primer.ConclusionsTherefore, using multiple 16S rRNA regions rather than one “universal” region can significantly improve the ability of microbial community reconstruction. In addition, we found that a short fragment is suitable for most genera identifications, and the proper conserved regions used for primer design are larger than before.