影响猪精子耐冻性的候选基因分析  

Analysis of candidate genes affecting semen freezability of boar sperm

作　　者：王文君[1] 王小康 徐国强 周翔[1,2] 刘榜 WANG Wenjun;WANG Xiaokang;XU Guoqiang;ZHOU Xiang;LIU Bang(Key Laboratory of Agricultural Animal Genetics,Breeding and Reproduction,Ministry of Education,Huazhong Agricultural University,Wuhan 430070,China;The Cooperative Innovation Center for Sustainable Pig Production/The Engineering Technology Research Center of Hubei Province Local Pig Breed Improvement,Wuhan 430070,China)

机构地区：[1]华中农业大学农业动物遗传育种与繁殖教育部重点实验室,武汉430070 [2]生猪健康养殖省部共建协同创新中心/湖北省地方猪品种改良工程技术研究中心,武汉430070

出　　处：《华中农业大学学报》2021年第3期203-211,共9页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金重点项目(31930104)。

摘　　要：在对通城猪精液冷冻保存的过程中,发现精液冻存质量存在个体差异。根据多次精液冻存后的精子活力检测结果,结合系谱关系筛选出精子耐冻与不耐冻个体各3头,并采用“中芯一号”SNP芯片进行群体遗传分化指数(FST)分析,应用生物信息学方法挖掘影响猪精子耐冻性差异的关键候选基因。结果显示:22头通城猪精液冻存后的精子活力存在明显差异,3头精子耐冻个体之间和3头精子不耐冻个体之间分别具有更近的亲缘关系,表明猪精子耐冻性差异可能受遗传因素影响;对耐冻性差异个体SNP芯片数据的FST分析结果显示,FST平均值为0.1705,存在较高程度的遗传分化;进一步选取FST前1%(FST>0.53)的266个SNP位点,并在其上下游100 kb区域共注释到了397个候选基因,富集在细胞形态、肌动蛋白细胞骨架和离子通道等生物学过程;其中KCNG4、ARPC1B、DNAAF4、MNS1、LIMK2、RPL31和VIM等基因可作为猪精子耐冻性差异的关键候选基因。以上结果说明,猪精子耐冻性受遗传因素影响,并受到关键基因控制,值得进一步对其基因功能进行研究。In the semen cryopreservation of Tongcheng boars,it was found that there were individual differences in semen quality after cryopreservation.According to the results of sperm motility after repeated semen cryopreservation and the pedigree relationship,three individuals with good semen freezability(GSF)and three individuals with poor semen freezability(PSF)were selected,respectively.Fixation index(FST)analysis of GSF and PSF was performed by using the"Zhongxin No.1"SNP chip,and the key candidate genes influencing the semen freezability were identified by bioinformatics method.The results showed that there were significant differences in sperm motility of 22 Tongcheng pigs after cryopreservation.The pedigree relationship among three GSF or three PSF was closer than that between them,which indicated that the differences of semen freezability in boar might be affected by genetic factor.After FST analysis,the mean value of FST was 0.1705,which indicated a high degree of genetic differentiation between GSF and PSF.Furthermore,266 SNPs in the top 1%of FST(FST>0.53)were selected.By annotating the 100 kb flank regions of these SNPs,we found 397 candidate genes,which enriched in the gene ontology(GO)terms related to cell morphology,actin cytoskeleton and ion channel,etc.Some highly differentiated genes in these biological processes such as KCNG4,ARPC1B,DNAAF4,MNS1,LIMK2,RPL31 and VIM were identified as key candidate genes.These results indicate that semen freezability of boar after cryopreservation is affected by genetic factor and controlled by some key genes,which are worthy of further study.

关 键 词：猪 精液冷冻保存 精子耐冻性 差异候选基因 SNP芯片 人工授精 

分 类 号：S828.2[农业科学—畜牧学]