基于16S rRNA高通量测序技术分析安格斯牛瘤胃微生物多样性和功能预测的研究

Functional Prediction of Rumen Microbial Diversity and Functions of Angus Cattle Based on 16S rRNA High-Throughput Sequencing

本研究旨在系统探索和分析安格斯牛瘤胃微生物多样性及其基因功能。选用体重550 kg左右的安格斯牛6头分成2组,利用基于16S rRNA高通量测序技术检测牛瘤胃液样本进行组学分析。结果表明,2组样本通过Illumina Miseq测序平台共获得86298条高质量优质序列,聚类为346个操作分类单元(OUT),经分类学鉴定分属13个门,21个纲,24个目,40个科,123个属。拟杆菌门(Bacteroidetes)43.16%和厚壁菌门(Firmicutes)36.29%为优势菌群。基于属的组成,依次为普雷沃氏菌属_7(Prevotella_7)29.28%、琥珀酸弧菌科_UCG-001(Succinivibrionaceae_UCG-001)11.30%、琥珀酸菌属(Succiniclasticum)11.10%、普雷沃氏菌属_1(Prevotella_1)6.65%、瘤胃球菌属_1(Ruminococcus_1)5.17%、琥珀酸弧菌属(Succinivibrio)2.75%等。16S功能预测和COG、KEGG代谢通路数据库对比发现,功能集中在氨基酸运输和代谢、碳水化合物转运及代谢的相关基因上,可能含有丰富的蛋白分解、转运及代谢酶相关基因和大量的纤维素以及木质素降解酶基因。经碳水化合物活性酶(CAZymes)注释和KEGG代谢酶数据库比对显示,预测的功能基因糖苷水解酶和糖基转移酶所占比例较高;产乙酸和丁酸相关酶基因丰度较高。安格斯牛瘤胃内含有丰富的蛋白质分解、木质纤维素降解和挥发性脂肪酸生成菌群及酶系,为展示瘤胃微生物多样性,发现和筛选新的功能酶基因提供参考。

This study was aimed at systematically explore and analysis the rumen microbial diversity and gene function of Angus cattle.Six Angus cattle with body weight at 550 kg were selected and used and divided into two groups for omics analysis of rumen fluid samples adopting 16S rRNA high-throughput sequencing technology.The results showed that:A total of 86,298 high-quality sequences were obtained from the two groups of samples using the Illumina Miseq sequencing platform,clustered into 346 operational taxonomic units(OTU),and identified by taxonomy as 13 phyla,21 classes,24 orders,40 family,and 123 genera.43.16%of Bacteroidetes and 36.29%of Firmicutes are dominant bacteria.Based on the composition of the genus,they were Prevotella_729.28%,Succinivibrionaceae_UCG-00111.30%,Succiniclasticum 11.10%,Prevotella_16.65%,Ruminococcus_15.17%,and Succinivibrio 2.75%.It was found by comparison of 16S functional prediction and COG,KEGG metabolic pathway databases that the functions were concentrated in amino acid transport and metabolism,carbohydrate transship and metabolism related genes,which may contain rich proteolysis,transship and metabolism enzyme related genes and a large number of fibers and lignin degrading enzyme genes.Comparison of the CAZymes annotation with the KEGG metabolizing enzyme database showed that the predicted functional genes glycoside hydrolase and glycosyltransferase accounted for a higher proportion;the acetogenic and butyric acid-related enzyme gene abundance was comparatively high.In sum,the rumen of Angus cattle is rich in bacterial population of protein degradation,lignocellulose degradation,and the formation of volatile fatty acid and its enzyme systems,and it laid theoretical foundation to display rumen microbial diversity,discover,and screen new functional enzyme genes.