首页|基于16S rRNA基因及宏基因组测序解析藏猪肠道菌群组成特征

基于16S rRNA基因及宏基因组测序解析藏猪肠道菌群组成特征

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[目的]旨在通过分析藏猪肠道菌群组成及其功能注释,探讨藏猪肠道菌群与其耐粗饲和抗逆性能的关系。[方法]以66头藏猪粪便微生物为试验材料,提取DNA样本,利用16S rRNA基因测序技术分析在门水平和属水平藏猪的肠道菌群组成,构建共有核心菌的菌群互作网络,结合27头藏猪肠道微生物宏基因组测序数据,进行KEGG通路和碳水化合物代谢酶(CAZymes)数据库注释分析,以期阐明与藏猪纤维代谢能力强和抗病力等优良特性相关的功能通路。[结果](1)16S rRNA测序分析结果显示,拟杆菌门(Bacteroidetes)(39。34%~60。72%)和厚壁菌门(Firmicutes)(24。65%~38。5%)在藏猪肠道微生物门水平上占优势,而在属水平则依次是普雷沃氏菌属(Prevotella)、密螺旋体属(Treponema)和琥珀酸弧菌属(Succinivibrio)的丰度最高;(2)利用Cytoscape软件构建藏猪肠道样品中共有的26个核心菌属之间的互作网络,各菌群在网络中存在合作关系和竞争关系,证明藏猪肠道菌群结构比较稳定;(3)宏基因组鸟枪法测序结果显示,在KEGG功能注释中藏猪肠道主要富集营养物质合成代谢和遗传信息处理相关通路,包括氨基酸的生物合成、碳代谢、群体感应通路等。此外,使用CAZymes数据库注释显示藏猪肠道中的糖基转移酶(GTs)基因家族相对丰度最高,主要为GT2、GT4和GT41基因,其次是糖苷水解酶基因家族(GHs)的GH13、GH2和GH3基因。[结论]藏猪具有丰富且独特的微生物组成和功能通路,与免疫和抗病相关的疣微菌门(Verrucomicrobia)和阿克曼氏菌(Akkermansia)是藏猪肠道特有的高丰度菌属,可能与藏猪适应高海拔低氧低温的环境及其抗病性能有关;与粗纤维代谢相关的密螺旋体属Treponema、Sphaerochaeta、纤维杆菌属Fibrobacter在藏猪肠道中富集,能够帮助藏猪降解饲粮中的粗纤维,适应半放牧的饲养模式。研究结果有助于了解肠道微生物在藏猪耐粗饲和抗逆性中的作用机制,为今后对藏猪优良特性的开发利用提供新的思路。
Elucidating gut microbial composition profiles of Tibetan pig based on 16S rRNA gene and metagenomic sequencing
[Objective]This study aims to investigate the relationship between the intestinal flora of Tibetan pigs and their roughage tolerance and resilience performance by analyzing the composition of intestinal flora and its functional annotation.[Method]Microbial DNA was extracted from the feces of 66 Tibetan pigs,and 16S rRNA gene sequencing was used to analyze the composition of the intestinal flora of Tibetan pigs at the phylum level and genus level.A flora interactions network of shared core bacteria was constructed.Combined with the metagenomic sequencing of intestinal microbes in 27 Tibetan pigs,the annotation analysis of the KEGG pathway and the carbohydrate metabolizing enzymes(CAZymes)database were performed to elucidate functional pathways related to the fiber metabolism and disease resistance in the Tibetan pigs.[Result](1)The results showed that in the study of intestinal flora composition of Tibetan pigs,16S rRNA sequencing analysis revealed that Bacteroidetes(39.34%-60.72%)and Firmicutes(24.65%-38.5%)were dominant at the level of intestinal microbial phylum,while at the genus level,Prevotella,Treponema and Succinivibrio had the highest abundance.(2)Cytoscape software was used to construct the interactions network between the 26 core genera shared in the Tibetan pig intestinal samples,and there were cooperative and competitive relationships among the groups in the network,which proved that the structure of the intestinal flora of Tibetan pigs was relatively stable.(3)Tibetan pig intestine was mainly enriched in pathways related to nutrient anabolism and genetic information processing in KEGG functional annotation based on the results of macrogenomic birdshot sequencing,including amino acid biosynthesis,carbon metabolism,and group sensing pathways.Furthermore,annotation using the CAZymes database showed the highest relative abundance of the glycosyltransferases(GTs)gene family in the intestine of Tibetan pigs,mainly GT2,GT4 and GT41 genes,followed by the glycoside hydrolase gene family(GHs)of GH13,GH2 and GH3 genes.[Conclusion]Tibetan pigs have rich and unique microbial composition and functional pathways.Verrucomicrobia and Akkermansia,which are related to immunity and disease resistance,are high abundance genera specific to the intestinal tract of Tibetan pigs,which may be related to the adaptation of Tibetan pigs to the environment of high altitude and low oxygen and low temperature and their disease resistance performance;those associated with crude fiber metabolism,such as Treponema,Sphaerochaeta and Fibrobacter are enriched in the intestinal tract of Tibetan pigs,which can help Tibetan pigs degrade crude fiber in the ration and adapt to the semi-grazing feeding mode.These results help us to understand the role of gut microbiota in good adaptability to roughage forage and harsh environmental stress of Tibetan pigs,and provide new ideas for the development and utilization of the excellent characteristics of Tibetan pigs in the future.

Tibetan pig16S rRNA genemetagenomic sequencinggut microbiotafunction capacity

李卓君、黄晓畅、柯善林、杨慧、周云燕、肖石军、陈从英、高军

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江西农业大学 猪遗传改良与种质创新全国重点实验室,江西 南昌 330045

南昌大学 抚州医学院,江西 抚州 344000

藏猪 16S rRNA基因 宏基因组测序 肠道菌群 肠道功能

国家自然科学基金地区科学基金项目

32260813

2024

江西农业大学学报
江西农业大学

江西农业大学学报

CSTPCD北大核心
影响因子:0.748
ISSN:1000-2286
年,卷(期):2024.46(5)