牦牛EPO、PPARα基因SNP与高原低氧适应的相关性分析

doi:10.16431/j.cnki.1671-7236.2021.05.014

摘要/Abstract

摘要： 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。χ²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。

关键词: 牦牛; EPO基因; PPARα基因; SNP; 高原低氧适应性

Abstract: The aim of this study was to explore the correlation between genetic polymorphism of erythropoietin (EPO), peroxisome proliferator-activated receptor α (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 PPARα genes 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 PPARα genes had 3 SNPs sites,in EPO gene there were rs527G→A,rs1031A→T and rs1192T→C,while in PPARα gene 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 PPARα gene was accorded with Hardy-Weinberg equilibrium in 6 yak groups (P>0.05).However,rs77471C→A and rs77534C→T of PPARα gene 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 PPARα gene was significantly higher than other haplotypes in 6 yak groups.Taken together,EPO and PPARα genes could be used as molecular markers for further exploring the mechanism of high altitude hypoxia adaptation of yak.

Key words: yak; EPO gene; PPARα gene; SNP; high altitude hypoxia adaptability

中图分类号:

S823.85

赵丽玲, 向超, 王会, 钟金城, 柴志欣, 王吉坤, 王嘉博, 武志娟, 信金伟, 姬秋梅. 牦牛EPO、PPARα基因SNP与高原低氧适应的相关性分析[J]. 中国畜牧兽医, 2021, 48(5): 1633-1646.

ZHAO Liling, XIANG Chao, WANG Hui, ZHONG Jincheng, CHAI Zhixin, WANG Jikun, WANG Jiabo, WU Zhijuan, XIN Jingwei, JI Qiumei. The Correlation Analysis Between EPO,PPARα genes SNP and High Altitude Hypoxia Adaptation in Yak[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1633-1646.

参考文献

[1] 施奇静,王静,孙军平,等.中国牦牛资源保护及可持续利用:驯化与品种培育[J].家畜生态学报,2016,37(1):81-86. SHI Q J,WANG J,SUN J P,et al.Conservation and sustainable utilization of yak resources of China:domestication and breed[J].Acta Ecologiae Animalis Domastici,2016,37(1):81-86.(in Chinese)
[2] 汪琦,钟金城,柴志欣,等.三江黄牛mtDNA D-Loop区遗传多样性及系统进化[J].西北农业学报,2016,25(9):1269-1278. WANG Q,ZHONG J C,CHAI Z X,et al.Analysis of genetic diversity of mtDNA D-Loop regions and phylogenetic evolution of Sanjiang cattle[J].Acta Agriculturae Boreali-occidentalis Sinica,2016,25(9):1269-1278.(in Chinese)
[3] 巴桑旺堆,郭仪,朱彦宾,等.牦牛高原低氧适应性生理及分子机制[J].世界生态学,2018,7(2):76-79. BA S W D,GUO Y,ZHU Y B,et al.Physiology and molecular mechanism of plateau adaptability in yak[J].International Journal of Ecology,2018,7(2):76-79.(in Chinese)
[4] 张剑搏,丁学智,ANUM ALI A,等.高原土著动物适应性进化的研究进展[J].畜牧兽医学报,2019,50(9):1723-1736. ZHANG J B,DING X Z,ANUM A A,et al.Advances in research on adaptive evolution of native animals of Tibetan plateau[J].Acta Veterinariaet Zootechnica Sinica,2019,50(9):1723-1736.(in Chinese)
[5] 党鹏举,李少斌,王继卿,等.甘肃高山细毛羊和湖羊HIF-2α基因遗传特性与高原低氧适应性分析[J].华北农学报,2016,31(3):88-93. DANG P J,LI S B,WANG J Q,et al.The analysis of Gansu Alpine Merino and Hu sheep HIF-2α gene genetic characteristics and adaptabilit[J].Acta Agriculturae Boreali-Sinica,2016,31(3):88-93.(in Chinese)
[6] LI C Y,LI X W,LIU J,et al.Investigation of the differences between the Tibetan and Han populations in the hemoglobin-oxygen affinity of red blood cells and in the adaptation to high-altitude environments[J].Hematology,2018,23(5):309-313.
[7] TOLINEN J P,HEIKKILA M,MALINEN M,et al.A long hypoxia-inducible factor 3 isoform 2 is a transcription activator that regulates erythropoietin[J].Cell Moleculor Life Science,2020,77(18):3627-3642.
[8] 赵生国,贺希文,郭永博,等.不同海拔猪种EPO基因核苷酸多态性研究[J].基因组学与应用生物学,2017,36(3):854-858. ZHAO S G,HE X W,GUO Y B,et al.Study onnucleotide polymorphisms of the erythropoietin (EPO) gene in pigs adapted to different altitudes[J].Genomics and Applied Biology,2017,36(3):854-858.(in Chinese)
[9] 张春梅,陈倩,谢雯,等.基于促红细胞生成素基因表达及血液生理生化指标探究湖羊在青海高海拔地区的适应性[J].动物营养学报,2020,32(11):5380-5387. ZHANG C M,CHEN Q,XIE W,et al.Based on erythropoietin gene expression and blood physio-logical and biochemical indexes to explore the adaptability of Hu sheep in high-altitude area[J].Chinese Journal of Animal Nutrition,2020,32(11):5380-5387.(in Chinese)
[10] URDAMPILLETA A,GONZALEZMUNIESA P,PORTILLO M P.Usefulness of combining intermittent hypoxia and physical exercise in the treatment of obesity[J].Journal of Physiology and Biochemistry,2012,68(2):289-304.
[11] SIMONSON T S,YANG Y Z,HUFF C D.Genetic evidence for high-altitude adaptation in Tibet[J].Science,2010,329(5987):72-75.
[12] 张亚强,刘秀,李少斌,等.藏绵羊PPARα基因遗传变异特征及组织表达分析[J].基因组学与应用生物学,2019,38(4):1519-1524. ZHANG Y Q,LIU X,LI S B,et al.Genetic variation characteristics and tissue expression analysis of PPARα gene in Tibetan sheep[J].Genomics and Applied Biology,2019,38(4):1519-1524.(in Chinese)
[13] 穷达,巴桑,唐晓惠,等.藏绵羊eNOS基因外显子8多态位点检测及高原适应性分析[J].高原农业,2019,3(3):277-281. QIONG D,BA S,TANG X H,et al.Thepolymorphisms of Tibet sheep eNOS gene exon 8 and its association with adaptability of plateau[J].Journal of Plateau Agriculture,2019,3(3):277-281.(in Chinese)
[14] 董坤哲.藏猪高海拔环境适应性分子机制探讨[D].北京:中国农业科学院,2015. DONG K Z.Molecular mechanism of high-altitude environmental adaptability of Tibetan pig[D].Beijing:Chinese Academy of Agricultural Sciences,2015.(in Chinese)
[15] 周长卿.牦牛MMP3、ADAM17、ARG2基因SNPs与其高原低氧适应性相关分析[D].兰州:甘肃农业大学,2014. ZHOU C Q.Relationship between yak MMP3,ADAM17,ARG2 gene SNPs and its adaptation to high altitude hypoxia[D].Lanzhou:Gansu Agricultural University,2014.(in Chinese)
[16] QIU Q,ZHANG G J,MA T,et al.The yak genome and adaptation to life at high altitude[J].Nature Genetic,2012,44(8):946-949.
[17] 金龙.藏猪高原适应性相关基因的鉴定及其DNA甲基化调控规律[D].雅安:四川农业大学,2015. JIN N.The identification and regulation of DNA methylation of adaptive genes in Tibetan pigs[D].Ya'an:Sichuan Agricultural University,2015.(in Chinese)
[18] WEI C,WANG H,LIU G,et al.Genome-wide analysis reveals adaptation to high altitudes in Tibetan sheep[J].Scientific Reports,2016,6:26770.
[19] 许海霞,吴建平,张利平,等.牦牛、黄牛和绵羊EPO基因部分片段遗传多态性分析[J].中国兽医学报,2013,33(6):950-955. XU H X,WU J P,ZHANG L P,et al.Analysis on genetic polymorphism of the partial fragment of EPO gene in yak,cattle and sheep[J].Chinese Journal of Veterinary Science,2013,33(6):950-955.(in Chinese)
[20] 陈雪梅,金双,钟金城,等.牦牛EPO基因的PCR-SSCP多态性研究[J].河南农业科学,2008,7:104-107. CHEN X M,JIN S,ZHONG J C,et al.Study of polymorphism on EPO gene in yak by PCR-SSCP[J].Journal Henan Agricultural Sciences,2008,7:104-107.(in Chinese)
[21] 许海霞.高海拔和低海拔部分家畜促红细胞生成素(EPO)基因遗传多态性研究[D].兰州:甘肃农业大学,2012. XU H X.Study on genetic polymorphism of erythropoietin (EPO) gene in high and low altitude livestock[D].Lanzhou:Gansu Agricultural University,2012.(in Chinese)
[22] 张亚强.藏绵羊EGLN1和PPARA基因变异特征与高原低氧适应性探究[D].兰州:甘肃农业大学,2018. ZHANG Y Q.Study on the EGLN1 and PPARA gene variation characteristics of Tibetan sheep and the adaptability of high altitude hypoxia[D].Lanzhou:Gansu Agricultural University,2018.(in Chinese)
[23] 张琼琼.牛PPARα、PPARβ基因多态性及其与生长性状相关性分析[D].信阳:信阳师范学院,2016. ZHANG Q Q.Polymorphism of bovine PPARα、PPARβ gene and its correlation with growth traits[D].Xinyang:Xinyang Normal College,2016.(in Chinese)