不同猪种ADIPOQ基因外显子2多态性及其与山西白猪体重和体尺性状的关联分析
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摘要
本研究旨在检测猪脂联素(adiponectin,ADIPOQ)基因外显子2的多态性,并分析其对山西白猪体重和体尺性状的影响。采用PCR-SSCP技术检测了长白猪、大白猪、杜洛克猪、山西白猪、山西黑猪和马身猪6个猪种392个个体ADIPOQ基因外显子2的多态性,并采用GLM程序分析了ADIPOQ基因外显子2多态性与山西白猪体重和体尺性状的关联性。结果显示,在ADIPOQ基因外显子2的89bp处检测到G→A错义突变,引起缬氨酸(Val)向异亮氨酸(Ile)的转变。ADIPOQ基因外显子2存在3种基因型:AA、AB、BB,2个等位基因:A和B。杜洛克猪中只有BB基因型,长白猪、大白猪、山西白猪和山西黑猪中BB基因型为优势基因型,马身猪中AA基因型频率最高。在引入品种长白猪、大白猪和杜洛克猪中B等位基因为优势等位基因,基因频率分别为0.96、0.96和1.00;在地方品种马身猪中A等位基因频率(0.52)略高于B等位基因(0.48);在培育品种山西白猪和山西黑猪中B等位基因频率分别为0.76和0.78,介于引入猪种和地方品种之间。基因型频率分布在马身猪、山西白猪和山西黑猪之间无显著差异(P>0.05),杜洛克猪与长白猪、大白猪间差异均不显著(P>0.05),而长白猪和大白猪间差异显著(P<0.05),任意一个引入品种与马身猪、山西白猪和山西黑猪之间的差异均达到极显著水平(P<0.01)。ADIPOQ基因外显子2多态性对断奶重有显著影响,其中BB基因型个体28日龄断奶重显著高于AA和AB基因型(P<0.05),AA和AB基因型间无显著差异(P>0.05),但对其他性状无显著影响,说明该位点只在个体发育早期阶段起作用。
This study was aimed to investigate the polymorphism of adiponectin(ADIPOQ)gene exon 2 and its correlation on body weights and body sizes in Shanxi White pig.A total of 392 pigsfrom Landrace pig,Large White pig,Duroc pig,Shanxi White pig,Shanxi Black pig and Mashen pig populations were used to detect the polymorphism of ADIPOQgene exon 2 by PCR-SSCP,and the association of the SNPs on body weights and body sizes in Shanxi White pig was analyzed by GLM.The results showed that there was a missense mutation of G→A at 89 bp of ADIPOQ gene exon 2,which resulted in the change of amino acid from valine(Val)to isoleucine(Ile).There were three genotypes(AA,AB and BB)and two alleles(A and B)at this site.There was only BB genotype in Duroc pig,and the advantageous genotype was also BB genotype in Landrace pig,Large White pig,Shanxi White pig and Shanxi Black pig,while AA genotype frequency in Mashen pig was greater than AB and BB genotypes.In the imported pig breeds such as Landrace pig,Large White pig and Duroc pig,the frequency of the dominant allele B were 0.96,0.96,and1.00,respectively.In Chinese domestic Mashen pig,the frequency of allele A(0.52)was little greater than that of allele B(0.48).The frequency of allele B in Shanxi White pig and Shanxi Black pig were 0.76 and 0.78,respectively.The genotype distribution among Mashen pig,Shanxi White pig and Shanxi Black pig was not significant(P>0.05),there was no significant difference between Duroc pig and Landrace pig or Duroc pig and Large White pig(P>0.05),however,the difference of genotype distribution between Landrace pig and Large White pig was significant(P<0.05).Comparing to the imported breeds with Chinese domestic Mashen pig and the composite populations of Shanxi White pig and Shanxi Black pig,the genotype distribution difference of each other was extremely significant(P<0.01).The effect of the SNP in exon 2 of ADIPOQ gene on weaning weight was significant,the weaning weight at 28-day-old of BB genotype individuals was significantly greater than those of AA and AB genotypes(P<0.05),and there was no significant difference between AA and AB genotypes(P>0.05).The effect of this polymorphic site on other traits was not significant,which indicated that it mainly played role at early stage of pig growth and development.
This study was aimed to investigate the polymorphism of adiponectin(ADIPOQ)gene exon 2 and its correlation on body weights and body sizes in Shanxi White pig.A total of 392 pigsfrom Landrace pig,Large White pig,Duroc pig,Shanxi White pig,Shanxi Black pig and Mashen pig populations were used to detect the polymorphism of ADIPOQgene exon 2 by PCR-SSCP,and the association of the SNPs on body weights and body sizes in Shanxi White pig was analyzed by GLM.The results showed that there was a missense mutation of G→A at 89 bp of ADIPOQ gene exon 2,which resulted in the change of amino acid from valine(Val)to isoleucine(Ile).There were three genotypes(AA,AB and BB)and two alleles(A and B)at this site.There was only BB genotype in Duroc pig,and the advantageous genotype was also BB genotype in Landrace pig,Large White pig,Shanxi White pig and Shanxi Black pig,while AA genotype frequency in Mashen pig was greater than AB and BB genotypes.In the imported pig breeds such as Landrace pig,Large White pig and Duroc pig,the frequency of the dominant allele B were 0.96,0.96,and1.00,respectively.In Chinese domestic Mashen pig,the frequency of allele A(0.52)was little greater than that of allele B(0.48).The frequency of allele B in Shanxi White pig and Shanxi Black pig were 0.76 and 0.78,respectively.The genotype distribution among Mashen pig,Shanxi White pig and Shanxi Black pig was not significant(P>0.05),there was no significant difference between Duroc pig and Landrace pig or Duroc pig and Large White pig(P>0.05),however,the difference of genotype distribution between Landrace pig and Large White pig was significant(P<0.05).Comparing to the imported breeds with Chinese domestic Mashen pig and the composite populations of Shanxi White pig and Shanxi Black pig,the genotype distribution difference of each other was extremely significant(P<0.01).The effect of the SNP in exon 2 of ADIPOQ gene on weaning weight was significant,the weaning weight at 28-day-old of BB genotype individuals was significantly greater than those of AA and AB genotypes(P<0.05),and there was no significant difference between AA and AB genotypes(P>0.05).The effect of this polymorphic site on other traits was not significant,which indicated that it mainly played role at early stage of pig growth and development.
引文
[1]唐金凤,杨颖,汪启迪,等.血清脂联素水平与肥胖度的关系[J].中华内分泌代谢杂志,2003,19(3):166-168.TANG J F,YANG Y,WANG Q D,et al.Relationship between serum adiponectin concentration and adiposity[J].Chinese Journal of Endocrinology and Metabolism,2003,19(3):166-168.(in Chinese)
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[5] HUSSAIN M K,DELI F A,ALGENBAI A H A,et al.Adiponectin gene polymorphisms as a predictor for development of type 2diabetes mellitus in Iraqi population[J].Gene,2018,662:118-122.
[6] HSUEH Y M,CHEN W J,LIN Y C,et al.Adiponectin gene polymorphisms and obesity increase the susceptibility to arsenic-related renal cell carcinoma[J].Toxicology and Applied Pharmacology,2018,350:11-20.
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[8] YAMAUCHI T,KAMON J,MINOKOSHI Y,et al.Adiponectin stimulates glucose utilization and fattyacid oxidation by activating AMP-activated protein kinase[J].Nature Medicine,2002,8(11):1288-1295.
[9]张佳,周杰,盛晟,等.重组脂联素(rAdp)对皖南花猪骨骼肌卫星细胞脂联素及受体和肌球蛋白重链基因表达的影响[J].农业生物技术学报,2011,19(2):335-341.ZHANG J,ZHOU J,SHENG S,et al.Effect of recombinant adiponectin(rAdp)on adiponectin,adiponectin receptors and myosin heavy chain(MyHC)mRNA expression in muscle satellite cell of Wannanhua pigs in vitro[J].Journal of Agricultural Biotechnology,2011,19(2):335-341.(in Chinese)
[10]盛晟,周杰,张佳,等.重组脂联素对皖南花猪脂肪细胞脂联素及其受体mRNA表达的影响[J].中国兽医学报,2012,32(4):577-582.SHENG S,ZHOU J,ZHANG J,et al.Effect of recombinate adiponectin and its receptors mRNA expression in adipocyte of Wannanhua pigs in vitro[J].Chinese Journal of Veterinary Science,2012,32(4):577-582.(in Chinese)
[11] HEINZ J F,SINGH S P,JANOWITZ U,et al.Characterization of adiponectin concentrations and molecular weight forms in serum,seminal plasma,and ovarian follicular fluid from cattle[J].Theriogenology,2015,83(3):326-333.
[12]钟志君,洪亮,顾以韧,等.藏猪和大约克夏猪血清中脂代谢相关细胞因子的发育性变化研究[J].养猪,2016,1:65-67.ZHONG Z J,HONG L,GU Y R,et al.The research on developmental changes of lipid metabolism related cytokines in serum of Tibetan and Yorkshire pigs[J].Swine Production,2016,1:65-67.(in Chinese)
[13]苗志国,谢红兵,王永强,等.淮南猪与长白猪脂肪代谢酶及脂联素的发育差异[J].贵州农业科学,2018,46(1):61-63.MIAO Z G,XIE H B,WANG Y Q,et al.Development difference in fat metabolic enzymes and serum adiponectin between Huainan and Landrace pig[J].Guizhou Agricultural Sciences,2018,46(1):61-63.(in Chinese)
[14] TURECK L V,LEITE N,SOUZA R L,et al.ADIPOQ single nucleotide polymorphism:Association with adiponectin and lipoproteins levels restricted to men[J].Meta Gene,2015,5:98-104.
[15] ZHOU Q,CHEN B,JI T,et al.Association of genetic variants in RETN,NAMPTand ADIPOQgene with glycemic,metabolic traits and diabetes risk in a Chinese population[J].Gene,2018,642:439-446.
[16] OLUSEGUN E,KHALID O,ABRAHAM A,et al.Association of adiponectin gene(ADIPOQ)polymorphisms with measures of obesity in Nigerian young adults[J].The Egyptian Journal of Medical Human Genetics,2018,19(2):123-127.
[17] PERALTA-ROMERO J J,KARAM-ARAUJOC R,BURQUETE-GARCA A I,et al.ADIPOQand ADIPOR2gene polymorphisms:Association with overweight/obesity in Mexican children[J].Boletin Médico del Hospital Infantil de México,2015,72(1):26-33.
[18] CEPICA S,MASOPUST M,KNOLL A,et al.Linkage and RH mapping of the porcine adiponectin gene on chromosome 13[J].Animal Genetics,2005,36(3):276-277.
[19]李云,张凌齐,周杰,等.脂联素及其受体与GnRH和GnIH在皖南花猪下丘脑中的发育性表达及相关性研究[J].西北农林科技大学学报(自然科学版),2013,41(12):7-13.LI Y,ZHANG L Q,ZHOU J,et al.Developmental expression and correlation analysis of adiponectin,adiponectin receptors,GnRHand GnIHin hypothalamus of Wannan Hua pigs[J].Journal of Northwest A&F University(Nature Science Edition),2013,41(12):7-13.(in Chinese)
[20] WANG P,KO Y,LIU B,et al.The expression of porcine adiponectin and stearoyl coenzyme A desaturase genes in differentiating adipocytes[J].Asian-Australasian Journal of Animal Sciences,2004,17:588-593.
[21] STEFANIA D,ROBERTA D,LUCA B,et al.Study of porcine adiponectin(ADIPOQ)gene and association of a missense mutation with EBVs for production and carcass traits in Italian Duroc heavy pigs[J].Livestock Science,2009,125:101-104.
[22]杨树猛,马登录,郭淑珍,等.藏羊脂联素基因多态性及其与产肉性能的相关性分析[J].中国草食动物科学,2013,34(2):5-9.YANG S M,MA D L,GUO S Z,et al.SNPs of adiponectin gene and its relationship with meat traits in Tibetan sheep[J].Chinese Herbivore Science,2013,34(2):5-9.(in Chinese)
[23] CHOI Y,DAVIS M,CHUNG H.Effects of genetic variants in the promoter region of the bovine adiponectin(ADIPOQ)gene on marbling of Hanwoo beef cattle[J].Meat Science,2015,105:57-62.
[24] DAI L,XIONG Y,DENG C,et al.Association of the A-G polymorphism in porcine adiponectin gene with fat deposition and carcass traits[J].Asian-Australasian Journal of Animal Sciences,2006,19:779-783.
[25]张依裕,徐琪,段修军,等.白羽番鸭脂联素基因SNP与肌内脂肪和血清总胆固醇的关联分析[J].中国畜牧杂志,2011,47(3):6-10.ZHANG Y Y,XU Q,DUAN X J,et al.SNP of adiponectin gene and its association with intramuscular fat and serum total cholesterol in White Muscovy[J].Chinese Journal of Animal Science,2011,47(3):6-10.(in Chinese)
[26]董飚,龚道清,孟和,等.鸭脂联素基因单核苷酸多态性检测及群体遗传分析[J].遗传,2007,29(8):995-1000.DONG B,GONG D Q,MENG H,et al.Identification and genetic analysis of SNPs in duck adiponectin gene[J].Hereditas,2007,29(8):995-1000.(in Chinese)
[27]杨彦杰,昝林森,王洪宝.秦川牛脂联素基因SNPs检测及其与胴体、肉质性状的相关性[J].遗传,2009,31(10):1006-1012.YANG Y J,ZAN L S,WANG H B.SNPs detection of adiponectin gene and its relationship with carcass and meat quality traits in Qinchuan cattle[J].Hereditas,2009,31(10):1006-1012.(in Chinese)
[28]凌飞,汪亮亮,杜红丽,等.猪脂联素基因启动子克隆及多态性研究[J].安徽农业科学,2010,38(1):413-415.LING F,WANG L L,DU H L,et al.Cloning of the porcine adiponectin promoter and study on genetic polymorphism[J].Journal of Anhui Agricultural Sciences,2010,38(1):413-415.(in Chinese)
[2] ZHANG Z,LI Y,YANG X,et al.Susceptibility of multiple polymorphisms in ADIPOQ,ADIPOR1and ADIPOR2genes to myocardial infarction in Han Chinese[J].Gene,2018,658:10-17.
[3]安平,王安平,母义明.脂联素与胰岛素抵抗研究进展[J].生物技术通讯,2017,28(3):360-365.AN P,WANG A P,MU Y M.Progress of the relationship between adiponectin and insulin resistance[J].Letters in Biotechnology,2017,28(3):360-365.(in Chinese)
[4] CHUNG H F,LONG K Z,HSU C C,et al.Adiponectin gene(ADIPOQ)polymorphisms correlate with the progression of nephropathy in Taiwanese male patients with type 2diabetes[J].Diabetes Research and Clinical Practice,2014,105(2):261-270.
[5] HUSSAIN M K,DELI F A,ALGENBAI A H A,et al.Adiponectin gene polymorphisms as a predictor for development of type 2diabetes mellitus in Iraqi population[J].Gene,2018,662:118-122.
[6] HSUEH Y M,CHEN W J,LIN Y C,et al.Adiponectin gene polymorphisms and obesity increase the susceptibility to arsenic-related renal cell carcinoma[J].Toxicology and Applied Pharmacology,2018,350:11-20.
[7] KADOWAKI T,YAMAUCHI T.Adiponectin and adiponectin receptors[J].Endocrine Reviews,2005,26(3):439-451.
[8] YAMAUCHI T,KAMON J,MINOKOSHI Y,et al.Adiponectin stimulates glucose utilization and fattyacid oxidation by activating AMP-activated protein kinase[J].Nature Medicine,2002,8(11):1288-1295.
[9]张佳,周杰,盛晟,等.重组脂联素(rAdp)对皖南花猪骨骼肌卫星细胞脂联素及受体和肌球蛋白重链基因表达的影响[J].农业生物技术学报,2011,19(2):335-341.ZHANG J,ZHOU J,SHENG S,et al.Effect of recombinant adiponectin(rAdp)on adiponectin,adiponectin receptors and myosin heavy chain(MyHC)mRNA expression in muscle satellite cell of Wannanhua pigs in vitro[J].Journal of Agricultural Biotechnology,2011,19(2):335-341.(in Chinese)
[10]盛晟,周杰,张佳,等.重组脂联素对皖南花猪脂肪细胞脂联素及其受体mRNA表达的影响[J].中国兽医学报,2012,32(4):577-582.SHENG S,ZHOU J,ZHANG J,et al.Effect of recombinate adiponectin and its receptors mRNA expression in adipocyte of Wannanhua pigs in vitro[J].Chinese Journal of Veterinary Science,2012,32(4):577-582.(in Chinese)
[11] HEINZ J F,SINGH S P,JANOWITZ U,et al.Characterization of adiponectin concentrations and molecular weight forms in serum,seminal plasma,and ovarian follicular fluid from cattle[J].Theriogenology,2015,83(3):326-333.
[12]钟志君,洪亮,顾以韧,等.藏猪和大约克夏猪血清中脂代谢相关细胞因子的发育性变化研究[J].养猪,2016,1:65-67.ZHONG Z J,HONG L,GU Y R,et al.The research on developmental changes of lipid metabolism related cytokines in serum of Tibetan and Yorkshire pigs[J].Swine Production,2016,1:65-67.(in Chinese)
[13]苗志国,谢红兵,王永强,等.淮南猪与长白猪脂肪代谢酶及脂联素的发育差异[J].贵州农业科学,2018,46(1):61-63.MIAO Z G,XIE H B,WANG Y Q,et al.Development difference in fat metabolic enzymes and serum adiponectin between Huainan and Landrace pig[J].Guizhou Agricultural Sciences,2018,46(1):61-63.(in Chinese)
[14] TURECK L V,LEITE N,SOUZA R L,et al.ADIPOQ single nucleotide polymorphism:Association with adiponectin and lipoproteins levels restricted to men[J].Meta Gene,2015,5:98-104.
[15] ZHOU Q,CHEN B,JI T,et al.Association of genetic variants in RETN,NAMPTand ADIPOQgene with glycemic,metabolic traits and diabetes risk in a Chinese population[J].Gene,2018,642:439-446.
[16] OLUSEGUN E,KHALID O,ABRAHAM A,et al.Association of adiponectin gene(ADIPOQ)polymorphisms with measures of obesity in Nigerian young adults[J].The Egyptian Journal of Medical Human Genetics,2018,19(2):123-127.
[17] PERALTA-ROMERO J J,KARAM-ARAUJOC R,BURQUETE-GARCA A I,et al.ADIPOQand ADIPOR2gene polymorphisms:Association with overweight/obesity in Mexican children[J].Boletin Médico del Hospital Infantil de México,2015,72(1):26-33.
[18] CEPICA S,MASOPUST M,KNOLL A,et al.Linkage and RH mapping of the porcine adiponectin gene on chromosome 13[J].Animal Genetics,2005,36(3):276-277.
[19]李云,张凌齐,周杰,等.脂联素及其受体与GnRH和GnIH在皖南花猪下丘脑中的发育性表达及相关性研究[J].西北农林科技大学学报(自然科学版),2013,41(12):7-13.LI Y,ZHANG L Q,ZHOU J,et al.Developmental expression and correlation analysis of adiponectin,adiponectin receptors,GnRHand GnIHin hypothalamus of Wannan Hua pigs[J].Journal of Northwest A&F University(Nature Science Edition),2013,41(12):7-13.(in Chinese)
[20] WANG P,KO Y,LIU B,et al.The expression of porcine adiponectin and stearoyl coenzyme A desaturase genes in differentiating adipocytes[J].Asian-Australasian Journal of Animal Sciences,2004,17:588-593.
[21] STEFANIA D,ROBERTA D,LUCA B,et al.Study of porcine adiponectin(ADIPOQ)gene and association of a missense mutation with EBVs for production and carcass traits in Italian Duroc heavy pigs[J].Livestock Science,2009,125:101-104.
[22]杨树猛,马登录,郭淑珍,等.藏羊脂联素基因多态性及其与产肉性能的相关性分析[J].中国草食动物科学,2013,34(2):5-9.YANG S M,MA D L,GUO S Z,et al.SNPs of adiponectin gene and its relationship with meat traits in Tibetan sheep[J].Chinese Herbivore Science,2013,34(2):5-9.(in Chinese)
[23] CHOI Y,DAVIS M,CHUNG H.Effects of genetic variants in the promoter region of the bovine adiponectin(ADIPOQ)gene on marbling of Hanwoo beef cattle[J].Meat Science,2015,105:57-62.
[24] DAI L,XIONG Y,DENG C,et al.Association of the A-G polymorphism in porcine adiponectin gene with fat deposition and carcass traits[J].Asian-Australasian Journal of Animal Sciences,2006,19:779-783.
[25]张依裕,徐琪,段修军,等.白羽番鸭脂联素基因SNP与肌内脂肪和血清总胆固醇的关联分析[J].中国畜牧杂志,2011,47(3):6-10.ZHANG Y Y,XU Q,DUAN X J,et al.SNP of adiponectin gene and its association with intramuscular fat and serum total cholesterol in White Muscovy[J].Chinese Journal of Animal Science,2011,47(3):6-10.(in Chinese)
[26]董飚,龚道清,孟和,等.鸭脂联素基因单核苷酸多态性检测及群体遗传分析[J].遗传,2007,29(8):995-1000.DONG B,GONG D Q,MENG H,et al.Identification and genetic analysis of SNPs in duck adiponectin gene[J].Hereditas,2007,29(8):995-1000.(in Chinese)
[27]杨彦杰,昝林森,王洪宝.秦川牛脂联素基因SNPs检测及其与胴体、肉质性状的相关性[J].遗传,2009,31(10):1006-1012.YANG Y J,ZAN L S,WANG H B.SNPs detection of adiponectin gene and its relationship with carcass and meat quality traits in Qinchuan cattle[J].Hereditas,2009,31(10):1006-1012.(in Chinese)
[28]凌飞,汪亮亮,杜红丽,等.猪脂联素基因启动子克隆及多态性研究[J].安徽农业科学,2010,38(1):413-415.LING F,WANG L L,DU H L,et al.Cloning of the porcine adiponectin promoter and study on genetic polymorphism[J].Journal of Anhui Agricultural Sciences,2010,38(1):413-415.(in Chinese)