绵羊FSTL1基因组织表达及其多态性与产羔数之间的关系  

Tissue Expression and Polymorphism of Sheep(Ovis aries) FSTL1 Gene and Their Association with Litter Size

作　　者：周梅[1] 曹晓涵[1] 贺小云[1] 孙庆 狄冉[1] 胡文萍[1] 王翔宇[1] 张效生[2] 张金龙[2] 刘秋月[1] 储明星[1] ZHOU Mei;CAO Xiao-Han;HE Xiao-Yun;SUN Qing;DI Ran;HU Wen-Ping;WANG Xiang- Yu;ZHANG Xiao-Sheng;ZHANG Jin-Long;LIU Qiu-Yue;CHU Ming-Xing(Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;Tianjin Institute of Animal Sciences, Tianjin 300381, China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所/农业部动物遗传育种与繁殖重点实验室,北京100193 [2]天津市畜牧兽医研究所,天津300381

出　　处：《农业生物技术学报》2018年第5期801-810,共10页Journal of Agricultural Biotechnology

基　　金：国家转基因科技重大专项(No.2016ZX08009-003-006和No.2016ZX08010-005-003);国家自然科学基金(No.31772580);国家肉羊产业技术体系专项(No.CARS-38);中央级公益性科研院所基本科研业务费专项(No.Y2017JC24;No.2017ywf-zd-13;No.2013ywf-zd-1;No.2015ywf-zd-2和No.2015ywf-zd-8);中国农业科学院科技创新工程(No.ASTIPIAS13;No.CAAS-XTCX2016010-01-03;No.CAAS-XTCX2016010-03-03和No.CAAS-XTCX2016011-02-02);宁夏农林科学院科技创新先导资金项目(No.DWJLC-2016001);内蒙古自治区科技重大专项;农业科研杰出人才及其创新团队项目;国家万人计划科技创新领军人才项目;天津市科技计划项目(No.16ZXZYNC00050)

摘　　要：产羔数是绵羊(Ovis aries)最重要的经济性状之一,该性状受微效多基因控制,目前已经发现的与产羔数相关的基因依然屈指可数,对其进行挖掘一直是研究热点。为了验证通过全基因组重测序(whole genome re-sequencing,WGS)筛选出来的卵泡抑制素样蛋白1(follistatin-like protein 1,FSTL1)基因组织表达水平及其多态性与绵羊产羔数之间的关系,本研究利用半定量反转录-聚合酶链反应(semi-quantitative reverse transcription and polymerase chain reaction,sqRT-PCR)和实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术对FSTL1基因在不同产羔能力绵羊各组织中的表达进行研究,同时采用Sequenom Mass ARRAY~?SNP技术对FSTL1基因4个单核苷酸多态性位点(single nucleotide polymorphism sites,SNPs)分别在多羔绵羊品种(小尾寒羊,380只)和单羔绵羊品种(滩羊,苏尼特羊,萨福克羊,杜泊羊和草原型藏羊,共380只)中的多态性进行检测,并与小尾寒羊产羔数进行关联分析。结果表明:FSTL1基因在绵羊组织中呈广谱表达;qRT-PCR显示,FSTL1基因在苏尼特羊垂体、卵巢、输卵管、子宫体以及子宫角中的表达量均高于小尾寒羊,但并未达到差异显著水平(P>0.05),另外,其在++型小尾寒羊垂体、输卵管和子宫体中的表达均显著高于BB和B+型(P<0.05);FSTL1基因4个SNPs的基因型频率和等位基因频率在单、多羔品种间差异均达到显著水平(P<0.05);另外,4个SNPs在小尾寒羊中均表现为中度多态(0.25<PIC<0.5),而在其他绵羊品种中表现为部分中度(0.25<PIC<0.5)和部分低度多态(PIC<0.25);卡方适合性检验表明,4个SNPs在绝大部分绵羊品种中均处于Hardy-Weinberg平衡状态(P>0.05);关联分析表明,FSTL1基因4个SNPs多态性与小尾寒羊各胎产羔数均无显著相关(P>0.05),且4个SNPs突变纯合型的产羔数基本上都低于野生纯合型。本研究初步得出,FSTL1基因的表达水平与绵羊产羔数之间可能存在一定程度�Litter size is one of the most important economic traits in sheep （Ovis aries） which is controlled by minorgenes. While genes that have been found to be related to litter size are numbered, and it is still a hotspot to mine them. To investigate the tissue expression level and polymorphisms of follistatin-like protein 1 gene （FSTL1） which is found by whole genome re-sequencing （WGS） and its association with litter size in sheep, semi-quantitative reverse transcription and polymerase chain reaction （sqRT-PCR） and quantitative real-time PCR （qRT-PCR） were performed to investigate the expression of FSTL1 gene in sheep with different lambing abilities. Multiparous （380 Small Tail Han sheep） and uniparous （total 380 for Tan, Sunite, Suffolk, Dorper and Prairie Tibetan sheep） sheep breeds were selected and Sequenom MassARRAY？SNP assay was applied to genotype four single nucleotide polymorphism sites （SNPs） of FSTL1 gene. Then the association was analyzed between FSTL1 and litter size in Small Tail Han sheep. The results showed that FSTL1 expressed in all tissues selected. qRT-PCR indicated that the expression of FSTL1 had no significant difference between uniparous and multiparous sheep （P＞0.05）, but the expression in pituitary, ovary, oviduct, uterus body and uterine horn of uniparous Sunite sheep was higher than that in multiparous Small Tail Han sheep, and the expression in pituitary, oviduct and uterus body of ＋＋ was significantly higher than those in BB and B＋ （P＜0.05）. From genotyping, this study found the genotype frequencies and allele frequencies of the four SNPs were significantly different between uniparous and multiparous sheep （P＜0.05）. Population genetic analysis showed that four SNPs were at moderate polymorphism in Small Tail Han sheep （0.25＜PIC＜0.5） and partial at moderate （0.25＜PIC＜0.5） and partial at low polymorphism （PIC＜0.25） in other sheep breeds. The χ2 test indicated that, the four SNPs were under Hardy-Weinberg equi

关 键 词：绵羊  卵泡抑制素样蛋白1基因(FSTL1)  组织表达  分型  产羔数 

分 类 号：S826[农业科学—畜牧学]