中华鳖GHSR基因SNPs的筛选及生长性状的关联分析
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摘要
为探究GHSR基因多态性对中华鳖(Pelodiscus sinensis)生长相关性状的影响,采用直接测序法在GHSR基因5'侧翼和3'侧翼上筛选SNPs位点,共检测到5个单核苷酸多态性位点:A335T、G397T、A527G、A13482C和T13526A。随机选取同批繁殖的1冬龄200只中华鳖用直接测序法进行SNPs位点的分型,并分析与生长性状的相关性。检测结果显示,所有SNP位点均符合Hardy-Weinberg平衡状态(p>0.05)。方差分析显示,A336T位点的AT、TT基因型的体重、背甲长、背甲宽和裙边宽4项生长数据均显著高于AA基因型。A13482C位点的AC基因型的体重、背甲长、背甲宽和裙边宽4项数据均显著高于AA基因型(p<0.05)。研究表明,本实验在GHSR基因上获得的这些SNP位点可能影响着中华鳖的生长性状或与之紧密连锁,可为中华鳖分子辅助育种提供助力与参考。
To investigate the effect of polymorphism in GHSR gene on the growth correlation traits of Chinese soft-shell turtle(Pelodiscus sinensis), this study screened SNPS site from 5'UTR and 3'UTR of GHSR gene by direct sequencing method. We detected 5 SNPs: A335 T, G397 T, A527 G, A13482 C and T13526 A. 200 Chinese soft-shell turtles, which were bred in the same batch and were 1 winter age, were randomly selected. The genotype and gene frequency of these SNPs in 200 Chinese Soft-shell Turtles, which was used for association analysis with growth traits, were assayed by direct sequencing. The result showed that all the SNPs were in Hardy-Weinberg equilibrium(p>0.05). The variance analysis results showed that the growth data(body weight, body length, body width, body height, calipash length) of AT and TT genotype were significantly higher than those of AA genotype in locus A336T(p<0.05). The growth data(body weight, body length, body width, calipash length) of AC genotype were significantly higher than those of AA genotype in locus A13482C(p<0.05). The results suggested that these SNPs loci obtained from the GHSR gene in this study might link or be closely related to the growth performance of Chinese soft-shell turtle, it could provide reference and help to molecular assisted selection.
To investigate the effect of polymorphism in GHSR gene on the growth correlation traits of Chinese soft-shell turtle(Pelodiscus sinensis), this study screened SNPS site from 5'UTR and 3'UTR of GHSR gene by direct sequencing method. We detected 5 SNPs: A335 T, G397 T, A527 G, A13482 C and T13526 A. 200 Chinese soft-shell turtles, which were bred in the same batch and were 1 winter age, were randomly selected. The genotype and gene frequency of these SNPs in 200 Chinese Soft-shell Turtles, which was used for association analysis with growth traits, were assayed by direct sequencing. The result showed that all the SNPs were in Hardy-Weinberg equilibrium(p>0.05). The variance analysis results showed that the growth data(body weight, body length, body width, body height, calipash length) of AT and TT genotype were significantly higher than those of AA genotype in locus A336T(p<0.05). The growth data(body weight, body length, body width, calipash length) of AC genotype were significantly higher than those of AA genotype in locus A13482C(p<0.05). The results suggested that these SNPs loci obtained from the GHSR gene in this study might link or be closely related to the growth performance of Chinese soft-shell turtle, it could provide reference and help to molecular assisted selection.
引文
Chen T.G.,Yi H.F.,Zhang Y.K.,and Xiang Z.,2006,DNA marker technologies and their applications in animal genetics and breeding,Xumu Shichang(Stockbreeding Market),(8):47-50(陈天国,易华锋,张廷科,向钊,2006,DNA分子标记与动物遗传育种,畜牧市场,(8):47-50)
Dong Y.,and Li Q.,2016,Correlation analysis between Snps and growth traits of Apostichopus japonicas,Haiyang Huzao Tongbao(Transactions of Oceanology and Limnology),(2):49-58(董玉,李琪,2016,刺参SNP标记与生长性状的关联分析,海洋湖沼通报,(2):49-58)
Kumar R.,Gupta I.D.,and Verma A.,2016,Novel SNP identification in exon 3 of HSP90AA1 gene and their association with heat tolerance traits in Karan Fries(Bos taurus×Bos indicus)cows under tropical climatic condition,Tropical Animal Health and Production,48(4):735-740
Lei M.,Luo C.,Peng X.,Fang M.,Nie Q.,Zhang D.,Yang G.,and Zhang X.,2007,Polymorphism of growth-correlated genes associated with fatness and muscle fiber traits in chickens,Poultry Science,86(5):835-842
Li J.Q.,Bao Z.M.,Li L.,and Hu X.L.,2013,Development and characterization of EST-SNP in Chlamys farreri,Zhongguo Haiyang Daxue Xuebao(Periodical of Ocean University of China),43(1):56-63(李纪勤,包振民,李玲,胡晓丽,2013,栉孔扇贝EST-SNP标记开发及多态性分析,中国海洋大学学报,43(1):56-63)
Liu B.,Garcia E.A.,and Korbonits M.,2011,Genetic studies on the ghrelin,growth hormone secretagogue receptor(GHSR)and ghrelin O-acyl transferase(GOAT)genes,Peptides,32(11):2191-2207
Liu B.,Song T.W.,Luo W.X.,Cai H.F.,and Sun Y.Y.,2013,Study on relationship between polymorphism of GHSR gene and growth traits in Qianbei Ma goat,Guangdong Nongye Kexue(Guangdong Agricultural Sciences),40(22):161-165(刘彬,宋桃伟,罗卫星,蔡惠芬,孙岩岩,2013,GHSR基因多态性与黔北麻羊生长性状关联性分析,广东农业科学,40(22):161-165)
Miao L.,Li M.Y.,Pan N.,Mu F.S.,Hou H.H.,Wang L.S.,and Chen J.,2017,Cloning,tissue expression and SNP screening of MSTN gene in Acrossocheilus fasciatus,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),25(5):700-708(苗亮,李明云,潘娜,穆方申,侯红红,王礼闪,陈炯,2017,光唇鱼MSTN基因克隆、组织表达及SNP位点筛查,农业生物技术学报,25(5):700-708)
Pan Y.J.,ed.,2007,Shuichancidian,Shanghai Lexicographical Publishing House,Shanghai,China,pp.26-28(潘迎捷,主编,2007,水产辞典,上海辞书出版社,中国,上海,pp.26-28)
Pantel J.,Legendre M.,Cabrol S.,Hilal L.,Hajaji Y.,Morisset S.,Nivot S.,and Vie L.,2006,Loss of constitutive activity of the growth hormone secretagogue receptor in familial short stature,Journal of Clinical Investigation,116(3):760
Song T.W.,Cai H.F.,Luo W.X.,Liu R.Y.,Zhang Y.Y.,Sun Y.Y.,and Liu B.,2015,Association of GHSR and GHRL gene genetic variation with growth traits in two Guizhou goat breeds,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(1):140-153(宋桃伟,蔡惠芬,罗卫星,刘若余,张依裕,孙岩岩,刘彬,2015,两个贵州山羊品种GHSR和GHRL基因遗传变异及其与生长性状的关联性,中国农业科学,48(1):140-153)
Wang C.X.,Lu M.X.,Gao F.Y.,Liu Z.G.,Cao J.M.,Zhu H.P.,and Ke X.L.,2015,Screening of single nucleotide polymophisms(SNPs)related with growth in growth hormone secretagogue receptor gene(GHSR)of Nile Tilapia(Oreochromis niloticus),Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),23(6):762-771(王春晓,卢迈新,高风英,刘志刚,曹建萌,朱华平,可小丽,2015,尼罗罗非鱼生长激素促分泌素受体基因(GHSR)生长相关单核苷酸多态性(SNPs)位点的筛选,农业生物技术学报,23(6):762-771)
Wang P.C.,ed.,2000,Turtle of China,East China Normal Uni versity Press,Shanghai,China,pp.50-56(王培潮,主编,2000,中国的龟鳖,华东师范大学出版社,中国,上海,pp.50-56)
Xiao Z.H.,2012,Study on clinical efficacy of Simo Decoration on patients whit functional dyspepsia and influence of Ghrelin,GHSR of model mice expression,Dissertation for Ph.D.,Hunan University of Chinese Medicine,Supervisor:Liu B.Y.,pp.78-80(肖政华,2012,四磨汤对功能性消化不良患者的临床疗效及模型小鼠Ghrelin,GHSR表达的影响,博士学位论文,湖南中医药大学,导师:刘柏炎,pp.78-80)
Yang Z.C.,Niu C.J.,and Sun R.Y.,1999,Advances in studies of the Chinese soft-shelled turtle Trionyx sinensis biology,Dongwuxue Zazhi(Chinese Journal of Zoology),34(6):41-44(杨振才,牛翠娟,孙儒泳,1999,中华鳖生物学研究进展,动物学杂志,34(6):41-44)
Yu J.H.,Xia D.Q.,Yang H.,He Y.H.,and Wu T.T.,2003,Morphology of the progenies of Oreochromis aurea(♀)×Sinipperca chuatai(♂),Shuichan Xuebao(Journal of Fisheries of China),27(5):431-435(俞菊华,夏德全,杨弘,贺艳辉,吴婷婷,2003,奥利亚罗非鱼(♀)×鳜(♂)杂交后代的形态,水产学报,27(5):431-435)
Zhang B.,Chen H.,and Guo Y.,2009,Associations of polymorphism within the GHSR gene with growth traits in Nanyang cattle,Molecular Biology Reports,36(8):22-59
Zhang B.,2008,Studies on the genetic variations of ND5 and GHSR genes and its association with growth traits in six cattle breeds,Thesis for M.S.,Northwest A&F University,Supervisor:Chen H.,pp.3-5(张宝,2008,6个黄牛品种ND5,GHSR基因遗传变异及其与生长性状关联分析,硕士学位论文,西北农林科技大学,导师:陈宏,pp.3-5)
Zhang J.,Ma W.,He Y.,Wu J.J.,Dawar F.U.,Ren F.,Zhao X.H.,and Mei J.,2016,Sex biased expression of ghrelin and GHSRassociated with sexual size dimorphism in yellow catfish,Gene,578(2):169-176
Zhu Z.M.,2013,Cloning,expression and SNPs analysis of two genes encoding lipoprotein lipases from Micropterus salmoides,Thesis for M.S.,Jinan University,Supervisor:Liang X.F.,pp.43-45(朱择敏,2013,大口黑鲈2种脂蛋白脂肪酶基因的克隆表达特征和SNP多态性分析,硕士学位论文,暨南大学,导师:梁旭方,pp.43-45)
Dong Y.,and Li Q.,2016,Correlation analysis between Snps and growth traits of Apostichopus japonicas,Haiyang Huzao Tongbao(Transactions of Oceanology and Limnology),(2):49-58(董玉,李琪,2016,刺参SNP标记与生长性状的关联分析,海洋湖沼通报,(2):49-58)
Kumar R.,Gupta I.D.,and Verma A.,2016,Novel SNP identification in exon 3 of HSP90AA1 gene and their association with heat tolerance traits in Karan Fries(Bos taurus×Bos indicus)cows under tropical climatic condition,Tropical Animal Health and Production,48(4):735-740
Lei M.,Luo C.,Peng X.,Fang M.,Nie Q.,Zhang D.,Yang G.,and Zhang X.,2007,Polymorphism of growth-correlated genes associated with fatness and muscle fiber traits in chickens,Poultry Science,86(5):835-842
Li J.Q.,Bao Z.M.,Li L.,and Hu X.L.,2013,Development and characterization of EST-SNP in Chlamys farreri,Zhongguo Haiyang Daxue Xuebao(Periodical of Ocean University of China),43(1):56-63(李纪勤,包振民,李玲,胡晓丽,2013,栉孔扇贝EST-SNP标记开发及多态性分析,中国海洋大学学报,43(1):56-63)
Liu B.,Garcia E.A.,and Korbonits M.,2011,Genetic studies on the ghrelin,growth hormone secretagogue receptor(GHSR)and ghrelin O-acyl transferase(GOAT)genes,Peptides,32(11):2191-2207
Liu B.,Song T.W.,Luo W.X.,Cai H.F.,and Sun Y.Y.,2013,Study on relationship between polymorphism of GHSR gene and growth traits in Qianbei Ma goat,Guangdong Nongye Kexue(Guangdong Agricultural Sciences),40(22):161-165(刘彬,宋桃伟,罗卫星,蔡惠芬,孙岩岩,2013,GHSR基因多态性与黔北麻羊生长性状关联性分析,广东农业科学,40(22):161-165)
Miao L.,Li M.Y.,Pan N.,Mu F.S.,Hou H.H.,Wang L.S.,and Chen J.,2017,Cloning,tissue expression and SNP screening of MSTN gene in Acrossocheilus fasciatus,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),25(5):700-708(苗亮,李明云,潘娜,穆方申,侯红红,王礼闪,陈炯,2017,光唇鱼MSTN基因克隆、组织表达及SNP位点筛查,农业生物技术学报,25(5):700-708)
Pan Y.J.,ed.,2007,Shuichancidian,Shanghai Lexicographical Publishing House,Shanghai,China,pp.26-28(潘迎捷,主编,2007,水产辞典,上海辞书出版社,中国,上海,pp.26-28)
Pantel J.,Legendre M.,Cabrol S.,Hilal L.,Hajaji Y.,Morisset S.,Nivot S.,and Vie L.,2006,Loss of constitutive activity of the growth hormone secretagogue receptor in familial short stature,Journal of Clinical Investigation,116(3):760
Song T.W.,Cai H.F.,Luo W.X.,Liu R.Y.,Zhang Y.Y.,Sun Y.Y.,and Liu B.,2015,Association of GHSR and GHRL gene genetic variation with growth traits in two Guizhou goat breeds,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),48(1):140-153(宋桃伟,蔡惠芬,罗卫星,刘若余,张依裕,孙岩岩,刘彬,2015,两个贵州山羊品种GHSR和GHRL基因遗传变异及其与生长性状的关联性,中国农业科学,48(1):140-153)
Wang C.X.,Lu M.X.,Gao F.Y.,Liu Z.G.,Cao J.M.,Zhu H.P.,and Ke X.L.,2015,Screening of single nucleotide polymophisms(SNPs)related with growth in growth hormone secretagogue receptor gene(GHSR)of Nile Tilapia(Oreochromis niloticus),Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),23(6):762-771(王春晓,卢迈新,高风英,刘志刚,曹建萌,朱华平,可小丽,2015,尼罗罗非鱼生长激素促分泌素受体基因(GHSR)生长相关单核苷酸多态性(SNPs)位点的筛选,农业生物技术学报,23(6):762-771)
Wang P.C.,ed.,2000,Turtle of China,East China Normal Uni versity Press,Shanghai,China,pp.50-56(王培潮,主编,2000,中国的龟鳖,华东师范大学出版社,中国,上海,pp.50-56)
Xiao Z.H.,2012,Study on clinical efficacy of Simo Decoration on patients whit functional dyspepsia and influence of Ghrelin,GHSR of model mice expression,Dissertation for Ph.D.,Hunan University of Chinese Medicine,Supervisor:Liu B.Y.,pp.78-80(肖政华,2012,四磨汤对功能性消化不良患者的临床疗效及模型小鼠Ghrelin,GHSR表达的影响,博士学位论文,湖南中医药大学,导师:刘柏炎,pp.78-80)
Yang Z.C.,Niu C.J.,and Sun R.Y.,1999,Advances in studies of the Chinese soft-shelled turtle Trionyx sinensis biology,Dongwuxue Zazhi(Chinese Journal of Zoology),34(6):41-44(杨振才,牛翠娟,孙儒泳,1999,中华鳖生物学研究进展,动物学杂志,34(6):41-44)
Yu J.H.,Xia D.Q.,Yang H.,He Y.H.,and Wu T.T.,2003,Morphology of the progenies of Oreochromis aurea(♀)×Sinipperca chuatai(♂),Shuichan Xuebao(Journal of Fisheries of China),27(5):431-435(俞菊华,夏德全,杨弘,贺艳辉,吴婷婷,2003,奥利亚罗非鱼(♀)×鳜(♂)杂交后代的形态,水产学报,27(5):431-435)
Zhang B.,Chen H.,and Guo Y.,2009,Associations of polymorphism within the GHSR gene with growth traits in Nanyang cattle,Molecular Biology Reports,36(8):22-59
Zhang B.,2008,Studies on the genetic variations of ND5 and GHSR genes and its association with growth traits in six cattle breeds,Thesis for M.S.,Northwest A&F University,Supervisor:Chen H.,pp.3-5(张宝,2008,6个黄牛品种ND5,GHSR基因遗传变异及其与生长性状关联分析,硕士学位论文,西北农林科技大学,导师:陈宏,pp.3-5)
Zhang J.,Ma W.,He Y.,Wu J.J.,Dawar F.U.,Ren F.,Zhao X.H.,and Mei J.,2016,Sex biased expression of ghrelin and GHSRassociated with sexual size dimorphism in yellow catfish,Gene,578(2):169-176
Zhu Z.M.,2013,Cloning,expression and SNPs analysis of two genes encoding lipoprotein lipases from Micropterus salmoides,Thesis for M.S.,Jinan University,Supervisor:Liang X.F.,pp.43-45(朱择敏,2013,大口黑鲈2种脂蛋白脂肪酶基因的克隆表达特征和SNP多态性分析,硕士学位论文,暨南大学,导师:梁旭方,pp.43-45)