基于16S rRNA高通量测序技术分析草原红牛瘤胃微生物多样性和功能预测的研究

Analysis of microbial diversitythe rumen of Grassland Red cattle and prediction of the function of the microbes based on the16S rRNA high-throughput sequencing technology

为系统探讨草原红牛瘤胃内的微生物多样性及其功能,本试验利用16S rRNA基因高通量测序技术检测分析草原红牛(20月龄左右,均重为577.5 kg)瘤胃液样本菌群结构并进行PICRUSt功能预测。结果显示:通过Illumina Miseq测序平台共获得35 848条优质序列,聚类分析得到387个操作分类单元(OTU),经分类学鉴定分属15个门、20个纲、25个目、41个科及110个属;厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)为优势菌群,所占比例分别为50.09%和41.11%;基于属的组成,依次为普雷沃菌属(Prevotella)15.20%、未知属f型拟杆菌目(norankfBacteroidales)BS11菌群8.66%、瘤胃菌科(Ruminococcaceae)NK4A214菌群6.96%、理研菌科(Rikenellaceae)RC9菌群5.56%、未知属(Christensenellaceae)R-7菌群4.01%、瘤胃球菌属2(Ruminococcus2)3.33%等;16S rRNA基因组的PICRUSt功能预测结果显示,瘤胃内菌群功能主要集中在碳水化合物转运及代谢,表面草原红牛体内含有大量的纤维素和木质素降解酶基因。综上,基于16S rRNA基因的高通量测序技术全面揭示了草原红牛瘤胃菌群的多样性,且预测其含有丰富的蛋白质分解、木质纤维素降解酶系,为探索草原红牛瘤胃微生物的认知提供了基础,也为挖掘其他重要营养生理功能相关的瘤胃微生物功能基因提供了参考。

To systematically explore the microbial diversity and the function of microbes in the rumen of Grassland Red cattle, the 16 S rRNA gene high-throughput sequencing technology was used to detect and analyze the flora structure of bacteria in the rumen fluid samples from Grassland Red cattle(of about 20 months old, and average weight of 577.5 kg) and PICRUSt was used to predict the function of themicrobes.The results were that a total of 35848 high-quality sequences were obtained through the Illumina Miseq sequencing platform, and 387 operational classification units(OTUs) were obtained by cluster analysis.The taxonomic identification showed that the microbes were of 15 phyla, 22 classes, 25 orders, and 41 families, and 110 genera.Firmicutes and Bacteroidetes wer dominant bacteria, accounting for 50.09% and 41.11%,respectively;and the compositions of the genus had the order of Prevotella: 15.20%,the norankfBacteroidalesBS11gutflorum: 8.66%,the RuminococcaceaeNK4 A214florum: 6.96%,the RikenellaceaeRC9gut florum: 5.56%,the ChristensenellaceaeR-7 florum: 4.01%,and the Ruminococcus2 florum: 3.33%.The prediction results the function of the 16 S r RNA genome by of PICRUSt showed that the function of the flora in the rumen was mainly to transport the carbohydrate and to perform metabolism,and the surface Grassland Red cattle contained a large amount of cellulose and lignin degrading enzyme genes.In summary,the high-throughput sequencing technology based on the 16 S r RNA gene fully revealed the diversity of bacteria in the rumen of Grassland Red cattle,and predicted that they were rich in protein decomposition and lignocellulose degrading enzymes,which provided reference for the knowledge of rumen microbes in Grassland Red cattle and was of help for exploring the biological functions of other microbes in rumen in terms of nutrition and physiology.