牦牛EPO、PPARα基因SNP与高原低氧适应的相关性分析

The Correlation Analysis Between EPO,PPARαgenes SNP and High Altitude Hypoxia Adaptation in Yak

试验旨在探究促红细胞生成素基因(EPO)、过氧化物酶体增殖剂激活受体α(PPARα)的遗传多样性,并分析其多态性与牦牛高原低氧适应性的相关性。采集不同海拔高度的6个牦牛类群(中甸牦牛、麦洼牦牛、斯布牦牛、类乌齐牦牛、帕里牦牛、申扎牦牛)以及三江黄牛共375头耳样,提取DNA并分别构建DNA池,采用直接测序法结合PCR-RFLP检测分析EPO、PPARα基因的多态性,最后应用SHEsis软件统计分析候选基因SNPs与牦牛高原适应性的相关性。结果表明,EPO基因存在3个SNPs位点:rs527G→A、rs1031A→T、rs1192T→C;PPARα基因存在3个SNPs位点:rs77363C→T、rs77471C→A和rs77534C→T。χ^(2)适合性检验结果显示,EPO基因3个SNPs位点均符合Hardy-Weinberg平衡状态;PPARα基因的rs77363C→T位点上,6个牦牛类群都处于平衡状态(P>0.05),在PPARα基因的rs77471C→A和rs77534C→T位点,麦洼牦牛偏离Hardy-Weinberg平衡(P<0.05)。单倍型分析得出,EPO基因的ATC单倍型在高海拔地区牦牛中的分布频率随海拔升高而升高;PPARα基因的TAC单倍型在6个牦牛类群中分布频率显著高于其他单倍型。研究表明,EPO、PPARα基因可作为牦牛适应高原环境的分子标记,为进一步探讨牦牛高原低氧适应性机制提供一定理论帮助。

The aim of this study was to explore the correlation between genetic polymorphism of erythropoietin(EPO),peroxisome proliferator-activated receptorα(PPARα)genes and hypoxia adaptability in yak.375 ears sample from 6 yak groups(Zhongdian yak,Maiwa yak,Sibu yak,Leiwuqi yak,Pali yak and Shenzha yak)and Sanjiang cattle at different altitudes were collected to extracted DNA and constructed DNA pool.The polymorphism of EPO and PPARαgenes were detected by direct sequencing and PCR-RFLP.Finally,the association between the SNPs of the candidate genes and the high altitude hypoxia adaptation of yak were analyzed by SHEsis.The results showed that both EPO and PPARαgenes had 3 SNPs sites,in EPO gene there were rs527G→A,rs1031A→T and rs1192T→C,while in PPARαgene the 3 SNPs sites were rs77363C→T,rs77471C→A and rs77534C→T.The fitness test indicated that the three SNPs sites of EPO gene all conformed to Hardy-Weinberg equilibrium;The rs77363C→T locus of PPARαgene was accorded with Hardy-Weinberg equilibrium in 6 yak groups(P>0.05).However,rs77471C→A and rs77534C→T of PPARαgene were deviated in Maiwa yak(P<0.05).The haplotype analysis showed that the distribution frequency of the ATC haplotype of EPO gene increased in high altitude yaks with the increase of altitude.Furthermore,the frequency of the TAC haplotype of PPARαgene was significantly higher than other haplotypes in 6 yak groups.Taken together,EPO and PPARαgenes could be used as molecular markers for further exploring the mechanism of high altitude hypoxia adaptation of yak.