基于Illumina MiSeq和IonS5 XL测序对原料乳中微生物多样性的比较分析

Comparative Analysis of Microbial Community in Raw Milk by Illumina MiSeq Sequencing and IonS5 XL Sequencing

不同的高通量测序平台各有优劣,针对研究对象选择合适的测序平台,对微生物多样性研究具有重要意义。以原料乳中的细菌为研究对象,采用Illumina MiSeq和IonS5 XL两种测序平台对细菌的V4—V5区进行扩增测序,探讨两种不同的测序平台在原料乳微生物多样性应用中的适用性。结果显示:两个测序平台的原始测序数据经质控后均可满足实验需求;IonS5 XL测序在不同分类水平上的注释物种数、alpha多样性、稀释曲线和物种丰度等级曲线方面的结果优于Illumina MiSeq测序,显示出原料乳中较高的群落丰富度;Illumina MiSeq测序和IonS5 XL测序均检出相同的优势菌门与优势菌属;Venn图和PCA分析显示,Illumina MiSeq测序相较于IonS5 XL测序具有更好的生物学重复性,在测序稳定性方面具有优势。后期如能加大平行样本的统计,针对重复性进行改进,IonS5 XL测序也将在乳品微生物多样性研究中具有很大的应用空间。

Different high-throughput sequencing platforms have their own advantages and disadvantages.An appropriate sequencing platform for the research object is of great significance to the study of microbial diversity.Taking the bacteria in raw milk as the research object,Illumina MiSeq and IonS5 XL sequencing platforms were used to amplify and sequence the V4-V5 region of bacteria,and the applicability of two different sequencing platforms in the microbial diversity of raw milk was discussed.The results showed that all of the original sequencing data from the two platforms could meet the experimental requirements after quality control.The results of IonS5 XL sequencing on the number of annotated species at different taxonomic levels,alpha diversity,rarefaction curve,and rank abundance curve were superior to those of Illumina MiSeq sequencing,indicating higher community abundance in raw milk.The same dominant phyla and dominant genera were found in both Illumina MiSeq sequencing and IonS5 XL sequencing.The Venn plots and PCA analysis showed that Illumina MiSeq sequencing had better biological repeatability than IonS5 XL sequencing and had advantages in sequencing stability.If the statistics of parallel samples could be increased and the repeatability could be improved,IonS5 XL sequencing will have great application prospects in dairy microbial diversity research.