转IGF-1基因超细毛羊与非转基因羊肠道粪便菌群多样性及组成差异分析

Differences of the Intestinal Microbial Flora Diversity and Composition in IGF-1 Transgenic Superfine Wool Sheep and Non-transgenic Sheep

旨在探究超细毛羊转入IGF-1基因后是否会引起超细毛羊肠道微生物菌群群落改变而导致转基因羊生物安全存在隐患的问题。本研究在同一羊场随机选取临床健康羊3组,其中转IGF-1基因阳性组(GP组)41只(24母(GDF),17公(GPM)),转基因阴性羊(GN组)43只(25母(GNF),18公(GNM)),非转基因超细毛羊(NG组)29只(18母(NGF),11公(NGM))。取直肠粪样,应用IIlumina HiSeq高通量测序技术测定粪便样本中细菌的16S rRNA V3-V4区序列,分析转基因超细毛羊与非转基因羊间肠道粪样细菌群落组成。结果表明,共计得到17个门,32个纲,56个目,94个科,228个属及185个种;LEfSe分析显示,GPF组有10个生物标记物;GPM组有5个,均是反刍动物肠道菌群的常见定植菌;NGF组的生物标记物为拟杆菌BS11科;NGM组为厚壁菌门。均为反刍动物肠道菌群的主要优势菌。3组肠道菌群的共享菌分析显示,在门纲目科属种6个分类水平拥有共享菌比例高达91%~94%。本研究证实,转入IGF-1基因并未改变超细毛羊肠道菌群的整体结构分布,转基因羊具有生物安全及环境安全性。

The study aimed to explore the intestinal microbial community changes after transfering the IGF-1 gene into the superfine wool sheep,which would lead to potential problems in the biosafety of transgenic sheep.Forty one individuals in IGF-1 transgene positive group(GP group,female(GPF)24 and male(GPM)17),43 individuals in genetically modified negative group(GN group,female(GNF)25 and male(GNM)18)and 29 individuals in non-genetically modified group(NG group,female(NGF)18 and male(NGM)11)from clinical healthy sheep in the same field were randomly selected.The rectal fecal samples were collected.The IIlumina HiSeq high-throughput sequencing technology was used to determine bacterial 16S rRNA V3-V4 area sequence,and the bacteria community composition in fecal samples of transgenic superfine wool sheep and non-transgenic sheep was analyzed.There were 17 phyla,32 classes,56 orders,94 families,228 genera and 185 species.LEfSe analysis showed that there were 10 biomarkers in the GPF group,there were 5 biomarkers in GPM group,all of which were common colonization bacteria of ruminant intestinal flora.The biomarkers in NGF group was Bacteroides BS11 family,the biomarkers in NGM group was Firmicutes.They were the main dominant bacteria of ruminant intestinal flora.Analysis of shared bacteria of intestinal flora in 3 groups showed that the proportion of shared bacteria was as high as 91%-94%in 6 taxa of genus and family.It was confirmed that the introduction of IGF-1 gene did not change the overall distribution of intestinal flora in superfine wool sheep,and the transgenic sheep were biosafety and environmental safety.