努比亚山羊BMPR-IB基因多态性与其产羔性状的关联分析
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摘要
【目的】明确努比亚山羊骨形态发生蛋白受体-IB(BMPR-IB)基因多态性与其产羔性状的关联,为山羊育种寻找新的遗传分子标记。【方法】采用DNA池结合PCR产物直接测序技术检测努比亚山羊BMPR-IB基因全部编码区的变异情况,以时间飞行质谱列阵技术对候选SNP位点进行基因分型,并分析BMPR-IB基因多态性与产羔性状(产羔数、初生重和断奶重)的关联性。【结果】努比亚山羊BMPR-IB基因5'UTR第469位存在A→G碱基突变,内含子1第1664位和内含子9第11344位处存在G→A碱基突变。3个SNP位点(5'UTR 469~(th)、Intron1 1664~(th)和Intron9 11344~(th))存在3种基因型(GG型、GA型和AA型),对应的多态信息含量(PIC)分别为0.37、0.11和0.37,其中Intron1 1664~(th)位点的基因频率和基因型频率处于Hardy-Weinberg平衡状态(P>0.05),5'UTR 469~(th)和Intron9 11344~(th)位点则处于Hardy-Weinberg不平衡状态(P<0.05)。关联分析结果表明,努比亚山羊BMPR-IB基因5'UTR第469位碱基由A→G,其GG型母羊比AA型母羊的产羔数和子代平均初生重有所减少,但子代平均断奶重增加;内含子1第1664位碱基由G→A,其GA型母羊比GG型母羊的产羔数和子代平均初生重均有所增加,但子代平均断奶重减轻;内含子9第11344位碱基由G→A,其AA型母羊比GG型母羊产羔数少,但子代平均初生重和平均断奶重均增加。可见,努比亚山羊BMPR-IB基因的3个SNP位点对其产羔性状均无显著影响(P>0.05)。【结论】努比亚山羊BMPR-IB基因存在3个SNP位点(5'UTR 469~(th)、Intron11664~(th)和Intron9 11344~(th)),但对努比亚山羊产羔性状影响不显著,说明BMPR-IB基因对山羊的产羔性状具有一定种属特异性。
【Objective】The correlation between Nubian goat bone morphogenetic protein receptor-IB(BMPR-IB)gene polymorphism and its lambing traits was clarified to search new genetic molecular markers related to goat breeding.【Method】In this study,DNA pool and PCR products sequencing technique were used to detect the variation of whole coding region of Nubian goat BMPR-IB gene. Time flight mass spectrometry was applied to perform the genotype of candidate SNP loci,and analyze the relationship between polymorphism of BMPR-IB gene and goat lambing traits(lamb number,primary weight and weaning weight).【Result】There was a G→A base mutation at the 469~(th)position of 5'UTR of BMPR-IB gene of Nubian goat,a G→A base mutation at the 1664~(th)position of intron 1 and the 11343 thposition of intron9. The three SNP loci(5'UTR 469~(th),Intron1 1664~(th)and Intron9 11344~(th))had three genotypes(GG type,GA type and AAtype),the polymorphic information conten(tPIC)respectively were 0.37,0.11 and 0.37. The gene frequency and genotype frequency of Intron1 1664~(th)were in Hardy-Weinberg equilibrium state(P>0.05),5'UTR 469~(th)and Intron9 11344~(th) were in Hardy-Weinberg imbalance(P<0.05). Correlation analysis showed that the 5'UTR 469~(th)base of the BMPR-IB gene of Nubian goat was A→G,which resulted in a reduction of the lamb number and the average primary weight of the lamb in GG ewes compared with the AA ewes,but increase of the average weaning weight of the offspring. Intron1 1664~(th)base was mutated from G to A,while the lamb number and the average primary weight of the GA ewes were higher than those of the GG ewes,but the average weaning weight of the offspring was reduced. Intron9 11344~(th)base was mutated from G to A. The AA type ewes had fewer lambs than the GG ewes,and their offspring's average primary weight and average weaning weight increased.The three SNP loci of BMPR-IB gene of Nubian goat had no significant effects on its lambing traits(P>0.05). 【Conclusion】There are three SNP loci in the BMPR-IB gene of Nubian goats(5'UTR 469~(th),Intron11664~(th)and Intron9 11344~(th)),but the effect on the lamb traits of Nubian goats is not significant,indicating that the BMPRIB gene has species specificity on lamb traits for goats.
【Objective】The correlation between Nubian goat bone morphogenetic protein receptor-IB(BMPR-IB)gene polymorphism and its lambing traits was clarified to search new genetic molecular markers related to goat breeding.【Method】In this study,DNA pool and PCR products sequencing technique were used to detect the variation of whole coding region of Nubian goat BMPR-IB gene. Time flight mass spectrometry was applied to perform the genotype of candidate SNP loci,and analyze the relationship between polymorphism of BMPR-IB gene and goat lambing traits(lamb number,primary weight and weaning weight).【Result】There was a G→A base mutation at the 469~(th)position of 5'UTR of BMPR-IB gene of Nubian goat,a G→A base mutation at the 1664~(th)position of intron 1 and the 11343 thposition of intron9. The three SNP loci(5'UTR 469~(th),Intron1 1664~(th)and Intron9 11344~(th))had three genotypes(GG type,GA type and AAtype),the polymorphic information conten(tPIC)respectively were 0.37,0.11 and 0.37. The gene frequency and genotype frequency of Intron1 1664~(th)were in Hardy-Weinberg equilibrium state(P>0.05),5'UTR 469~(th)and Intron9 11344~(th) were in Hardy-Weinberg imbalance(P<0.05). Correlation analysis showed that the 5'UTR 469~(th)base of the BMPR-IB gene of Nubian goat was A→G,which resulted in a reduction of the lamb number and the average primary weight of the lamb in GG ewes compared with the AA ewes,but increase of the average weaning weight of the offspring. Intron1 1664~(th)base was mutated from G to A,while the lamb number and the average primary weight of the GA ewes were higher than those of the GG ewes,but the average weaning weight of the offspring was reduced. Intron9 11344~(th)base was mutated from G to A. The AA type ewes had fewer lambs than the GG ewes,and their offspring's average primary weight and average weaning weight increased.The three SNP loci of BMPR-IB gene of Nubian goat had no significant effects on its lambing traits(P>0.05). 【Conclusion】There are three SNP loci in the BMPR-IB gene of Nubian goats(5'UTR 469~(th),Intron11664~(th)and Intron9 11344~(th)),but the effect on the lamb traits of Nubian goats is not significant,indicating that the BMPRIB gene has species specificity on lamb traits for goats.
引文
焦丹,刘秀,李少斌,王继卿,闫伟,李文浩,罗玉柱.2016.绵羊BMPR1B基因的生物信息学分析[J].甘肃农业大学学报,51(4):1-6.[Jiao D,Liu X,Li S B,Wang J Q,Yan W,Li W H,Luo Y Z.2016.Bioinformatics analysis of BMPR1B gene in sheep[J].Journal of Gansu Agricultural University,51(4):1-6.]
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李文杨,刘远,林仕欣,张晓佩,高承芳,林碧芬,董晓宁.2012.BMPR-IB,BMP15,GDF9基因作为福建省4个主要山羊品种多胎性能候选基因的研究[J].浙江农业学报,24(6):975-979.[Li W Y,Liu Y,Lin S X,Zhang X P,Gao C F,Lin B F,Dong X N.2012.Studies on BMPR-IB,BMP15 and GDF9 as candidate genes of fecundity in four breeds of goats in Fujian[J].Acta Agriculturae Zhejiangensis,24(6):975-979.]
李隐侠,张俊,钱勇,孟春花,王慧利,钟声,曹少先.2017.湖羊NR5A1基因SNPs筛选及其与产羔数的关联分析[J].江苏农业学报,33(1):124-132.[Li Y X,Zhang J,Qian Y,Meng C H,Wang H L,Zhong S,Cao S X.2017.SNPs selection of NR5A1 and its association analysis with litter size in Hu sheep[J].Jiangsu Journal of Agricultural Sciences,33(1):124-132.]
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刘远,李文杨,林仕欣,张晓佩,高承芳,林碧芬,董晓宁.2012b.BMPR-IB、BMP15、GDF9基因作为闽东山羊多胎性能候选基因的研究[J].家畜生态学报,33(1):19-22.[Liu Y,Li W Y,Lin S X,Zhang X P,Gao C F,Lin B F,Dong X N.2012b.Study on BMPR-IB,BMP15 and GDF9 as candidate genes for fecundity in mindong goat[J].Acta Ecologae Animalis Domastici,33(1):19-22.]
潘章源,狄冉,刘秋月,储明星.2015.绵羊多羔主效基因BMPR1B的研究进展[J].家畜生态学报,36(5):1-6.[Pan Z Y,Di R,Liu Q Y,Chu M X.2015.Advances in ovine prolificacy gene BMPR1B[J].Journal of Domestic Animal Ecology,36(5):1-6.]
邵勇钢,米日尼萨汗·库尔班,刘武军.2012.策勒黑羊BMPR-IB基因多态性与产羔数的相关性研究[J].中国畜牧兽医,39(6):221-223.[Shao Y G,Mirinisahan Kuerban,Liu W J.2012.Study on the relationship between the polymorphism of BMPR-IB gene and litter size in Xinjiang Cele Black sheep[J].China Animal Husbandry&Veterinary Medicine,39(6):221-223.]
孙红霞,田秀娥,王永军.2009.BMPR-IB、BMP15和GDF9基因作为滩羊繁殖性状主效候选基因的研究[J].西北农业学报,18(5):17-21.[Sun H X,Tian X E,Wang Y J.2009.BMPR-IB,BMP15 and GDF9 as candidate genes for prolific trait in Tan sheep[J].Acta Agriculturae Boreali-Occidentalis Sinica,18(5):17-21.]
王小佳,贺建宁,程明,刘开东,刘积风,柳楠.2017.BMPR1B基因在细毛羊皮肤毛囊中的表达及与羊毛性状关系的研究[J].中国畜牧杂志,53(1):33-38.[Wang X J,He JN,Cheng M,Liu K D,Liu J F,Liu N.2017.Expression of BMPR1B gene in skin and its association with wool traits in fine wool sheep[J].Chinese Journal of Animal Science,53(1):33-38.]
姚雅馨.2017.绵羊FecB突变高通量分型技术的建立及巴美肉羊多羔候选基因的筛选[D].北京:中国农业科学院.[Yao Y X.2017.Establishment of high throughput genotyping methods for FecB mutation in sheep and screening for candidate genes of prolificacy in Bamei Mutton sheep[D].Beijing:Chinese Academy of Agricultural Sciences.]
邹辉,崔悦悦,夏琴,程晓芳,吴延军,张名媛,廖顺连,韦英明,蒋钦杨.2018.山羊产羔性状相关基因研究进展[J].黑龙江畜牧兽医,(11):63-67.[Zou H,Cui Y Y,Xia Q,Cheng X F,Wu Y J,Zhang M Y,Liao S L,Wei Y M,Jiang Q Y.2018.Research progress on the lambing traits related genes in goat[J].Heilongjiang Journal of Animal Science and Veterinary Medicine,(11):63-67.]
Chu M X,Zhao X H,Zhang Y J,Wang J Y,Di R,Cao G L,Feng T,Fang L,Ma Y H,Li K.2012.Polymorphisms of BMPR-IB gene and their relationship with litter size in goats[J].Molecular Biology Reports,37(8):4033-4039.
Dash S,Sarashetti P M,Rajashekar B,Chowdhury R,Mukherjee S.2018.TGF-β2-induced EMT is dampened by inhibition of autophagy and TNF-αtreatment[J].Oncotarget,9(5):6433-6449.
Mahdavi M,Nanekarani S,Hosseini S D.2014.Mutation in BMPR-IB gene is associated with litter size in Iranian Kalehkoohi sheep[J].Animal Reproduction Science,147(3-4):93-98.
Miao X,Luo Q,Qin X.2016.Genome-wide transcriptome analysis in the ovaries of two goats identifies differentially expressed genes related to fecundity[J].Gene,582(1):69-76.
Rodriguez A,Tripurani S K,Burton J C,Clementi C,Larina I,Pangas S A.2016.SMAD signaling is required for structural integrity of the female reproductive tract and uterine function during early pregnancy in mice[J].Biology of Reproduction,95(2):44.
Souza C J,MacDougall C,Campbell B K,Mcneilly A S,Baird D T.2001.The Booroola(FecB)phenotype is associated with a mutation in the bone morphogenetic receptor type 1 B(BMPR1B)gene[J].The Journal of Endocrinology,169(2):R1-R6.
Vacca G M,Dhaouadi A,Rekik M,Carcangiu V,Pazzola M,Dettori M L.2010.Prolificacy genotypes at BMPR1B,BMP15 and GDF9 genes in North African sheep breeds[J].Small Ruminant Research,88(1):67-71.
Wang W M,Liu S J,Li F,Pan X Y,Li C,Zhang X X,Ma YJ,La Y F,Xi R,Li T F.2015.Polymorphisms of the ovine BMPR-IB,BMP-15 and FSHR and their associations with litter size in two Chinese indigenous sheep breeds[J].International Journal of Molecular Sciences,16(5):11385-11397.
李闯,柳海星,高见红,乔佳琦.2018.羊主效多胎基因的研究现状[J].延边大学农学学报,40(1):87-93.[Li C,Liu HX,Gao J H,Qiao J Q.2018.A research progress on major prolificacy genes in sheep[J].Agricultural Science Journal of Yanbian University,40(1):87-93.]
李文杨,刘远,林仕欣,张晓佩,高承芳,林碧芬,董晓宁.2012.BMPR-IB,BMP15,GDF9基因作为福建省4个主要山羊品种多胎性能候选基因的研究[J].浙江农业学报,24(6):975-979.[Li W Y,Liu Y,Lin S X,Zhang X P,Gao C F,Lin B F,Dong X N.2012.Studies on BMPR-IB,BMP15 and GDF9 as candidate genes of fecundity in four breeds of goats in Fujian[J].Acta Agriculturae Zhejiangensis,24(6):975-979.]
李隐侠,张俊,钱勇,孟春花,王慧利,钟声,曹少先.2017.湖羊NR5A1基因SNPs筛选及其与产羔数的关联分析[J].江苏农业学报,33(1):124-132.[Li Y X,Zhang J,Qian Y,Meng C H,Wang H L,Zhong S,Cao S X.2017.SNPs selection of NR5A1 and its association analysis with litter size in Hu sheep[J].Jiangsu Journal of Agricultural Sciences,33(1):124-132.]
刘远,李文杨,林仕欣,张晓佩,高承芳,林碧芬,董晓宁.2012a.BMPR-IB、BMP15、GDF9基因作为福清山羊高繁殖力候选基因的研究[J].中国草食动物科学,32(3):12-14.[Liu Y,Li W Y,Lin S X,Zhang X P,Gao C F,Lin B F,Dong X N.2012a.Study on BMPR-IB,BMP15 and GDF9 as candidate genes for fecundity in Fuqing goat[J].China Herbivore Science,32(3):12-14.]
刘远,李文杨,林仕欣,张晓佩,高承芳,林碧芬,董晓宁.2012b.BMPR-IB、BMP15、GDF9基因作为闽东山羊多胎性能候选基因的研究[J].家畜生态学报,33(1):19-22.[Liu Y,Li W Y,Lin S X,Zhang X P,Gao C F,Lin B F,Dong X N.2012b.Study on BMPR-IB,BMP15 and GDF9 as candidate genes for fecundity in mindong goat[J].Acta Ecologae Animalis Domastici,33(1):19-22.]
潘章源,狄冉,刘秋月,储明星.2015.绵羊多羔主效基因BMPR1B的研究进展[J].家畜生态学报,36(5):1-6.[Pan Z Y,Di R,Liu Q Y,Chu M X.2015.Advances in ovine prolificacy gene BMPR1B[J].Journal of Domestic Animal Ecology,36(5):1-6.]
邵勇钢,米日尼萨汗·库尔班,刘武军.2012.策勒黑羊BMPR-IB基因多态性与产羔数的相关性研究[J].中国畜牧兽医,39(6):221-223.[Shao Y G,Mirinisahan Kuerban,Liu W J.2012.Study on the relationship between the polymorphism of BMPR-IB gene and litter size in Xinjiang Cele Black sheep[J].China Animal Husbandry&Veterinary Medicine,39(6):221-223.]
孙红霞,田秀娥,王永军.2009.BMPR-IB、BMP15和GDF9基因作为滩羊繁殖性状主效候选基因的研究[J].西北农业学报,18(5):17-21.[Sun H X,Tian X E,Wang Y J.2009.BMPR-IB,BMP15 and GDF9 as candidate genes for prolific trait in Tan sheep[J].Acta Agriculturae Boreali-Occidentalis Sinica,18(5):17-21.]
王小佳,贺建宁,程明,刘开东,刘积风,柳楠.2017.BMPR1B基因在细毛羊皮肤毛囊中的表达及与羊毛性状关系的研究[J].中国畜牧杂志,53(1):33-38.[Wang X J,He JN,Cheng M,Liu K D,Liu J F,Liu N.2017.Expression of BMPR1B gene in skin and its association with wool traits in fine wool sheep[J].Chinese Journal of Animal Science,53(1):33-38.]
姚雅馨.2017.绵羊FecB突变高通量分型技术的建立及巴美肉羊多羔候选基因的筛选[D].北京:中国农业科学院.[Yao Y X.2017.Establishment of high throughput genotyping methods for FecB mutation in sheep and screening for candidate genes of prolificacy in Bamei Mutton sheep[D].Beijing:Chinese Academy of Agricultural Sciences.]
邹辉,崔悦悦,夏琴,程晓芳,吴延军,张名媛,廖顺连,韦英明,蒋钦杨.2018.山羊产羔性状相关基因研究进展[J].黑龙江畜牧兽医,(11):63-67.[Zou H,Cui Y Y,Xia Q,Cheng X F,Wu Y J,Zhang M Y,Liao S L,Wei Y M,Jiang Q Y.2018.Research progress on the lambing traits related genes in goat[J].Heilongjiang Journal of Animal Science and Veterinary Medicine,(11):63-67.]
Chu M X,Zhao X H,Zhang Y J,Wang J Y,Di R,Cao G L,Feng T,Fang L,Ma Y H,Li K.2012.Polymorphisms of BMPR-IB gene and their relationship with litter size in goats[J].Molecular Biology Reports,37(8):4033-4039.
Dash S,Sarashetti P M,Rajashekar B,Chowdhury R,Mukherjee S.2018.TGF-β2-induced EMT is dampened by inhibition of autophagy and TNF-αtreatment[J].Oncotarget,9(5):6433-6449.
Mahdavi M,Nanekarani S,Hosseini S D.2014.Mutation in BMPR-IB gene is associated with litter size in Iranian Kalehkoohi sheep[J].Animal Reproduction Science,147(3-4):93-98.
Miao X,Luo Q,Qin X.2016.Genome-wide transcriptome analysis in the ovaries of two goats identifies differentially expressed genes related to fecundity[J].Gene,582(1):69-76.
Rodriguez A,Tripurani S K,Burton J C,Clementi C,Larina I,Pangas S A.2016.SMAD signaling is required for structural integrity of the female reproductive tract and uterine function during early pregnancy in mice[J].Biology of Reproduction,95(2):44.
Souza C J,MacDougall C,Campbell B K,Mcneilly A S,Baird D T.2001.The Booroola(FecB)phenotype is associated with a mutation in the bone morphogenetic receptor type 1 B(BMPR1B)gene[J].The Journal of Endocrinology,169(2):R1-R6.
Vacca G M,Dhaouadi A,Rekik M,Carcangiu V,Pazzola M,Dettori M L.2010.Prolificacy genotypes at BMPR1B,BMP15 and GDF9 genes in North African sheep breeds[J].Small Ruminant Research,88(1):67-71.
Wang W M,Liu S J,Li F,Pan X Y,Li C,Zhang X X,Ma YJ,La Y F,Xi R,Li T F.2015.Polymorphisms of the ovine BMPR-IB,BMP-15 and FSHR and their associations with litter size in two Chinese indigenous sheep breeds[J].International Journal of Molecular Sciences,16(5):11385-11397.
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