马MC1R基因多态位点与毛色相关性分析

doi:10.11843/j.issn.0366-6964.2018.08.005

摘要/Abstract

摘要：

旨在研究马MC1R基因多态位点与马毛色的关联性，为马的毛色育种提供重要的理论参考依据。本研究采集了10种不同毛色共132匹马的颈静脉血，提取基因组并应用PCR技术克隆MC1R基因编码序列，通过限制性长度多态性（RFLP）分析和测序分析，对马MC1R基因多态性与毛色进行关联分析，并预测了SNPs对其编码蛋白结构的影响。构建和分析了MC1R基因的系统发育树。结果发现，马MC1R基因存在3个多态位点，其中c.G102A为同义突变，c.C248T和c.G250A为错义突变。c.C248T造成MC1R蛋白的第83个氨基酸由极性亲水丝氨酸变为非极性疏水苯丙氨酸，预测突变后该位点氨基酸与第三跨膜区的第127个氨基酸（丝氨酸）之间的氢键消失而改变了MC1R的蛋白构象，该突变导致MC1R基因出现EE、Ee和ee 3种基因型。错义突变c.G250A导致其翻译的氨基酸由天冬氨酸变为天冬酰胺，该位点突变后与邻近氨基酸及跨膜域作用力复杂，推测比只出现c.C248T突变对MC1R蛋白功能的影响要小，该位点突变导致出现基因型ee^a。综合NCBI中的马MC1R基因型数据（共571匹）与其毛色性状进行相关性分析发现，在骝毛马和黑毛马群体中存在EE和Ee两种基因型，青毛马群体中存在EE、Ee、ee和ee^a基因型，栗毛中存在ee和ee^a两种基因型。黑毛和骝毛马的基因型分布差异极显著（P<0.01），呈现Hardy-Weinberg不平衡，等位基因E为优势等位基因；在栗色马中未检测到E等位基因，e为优势等位基因。MC1R基因系统发育树分析显示，EE基因型个体与家马、普氏野马相似性最高。本研究结果显示，马MC1R多态位点产生的不同基因型与马的骝毛、黑毛、栗毛及以这3种毛色为基础毛色的花马毛色存在相关性，骝毛马与黑毛马中不存在基因型ee，栗毛马中则不存在等位基因E。为应用基因型对马的毛色进行定向选择育种奠定了基础。

Abstract:

The study of the association between horse MC1R gene polymorphic loci and coat colors would provide important theoretical reference for coat color breeding of horses. In this study, the CDS of MC1R gene from 132 horses with 10 different coat colors was obtained by cloning and Polymerase Chain Reaction (PCR), Restriction Fragment Length Polymorphsm (RFLP) and sequencing were implemented to analyze the correlation between MC1R gene polymorphism and coat color, the effect of SNPs on its coding protein structure was predicted, and the MC1R gene NJ phylogenetic tree was constructed and analyzed. The results showed that 3 SNPs loci were found in horse MC1R gene, including one synonymous mutation(c.G102A) and two missense mutations(c.C248T, c.G250A).The c.C248T caused change of the 83rd amino acids of MC1R protein from polar hydrophilic serine to nonpolar hydrophobic phenylalanine, predicting that the hydrogen bond between amino acid at this locus and 127th amino acid (serine) at the third transmembrane domain would disappear and the conformation of MC1R protein would be changed. This mutation resulted in 3 genotypes of EE, Ee and ee in MC1R gene.The missense mutation c.G250A caused the change of amino acid from aspartic acid to asparagine. The mutation site had more complex interaction with adjacent amino acids and transmembrane domains. It was speculated that c.G250A and c.C248T had less effect on the MC1R protein function than only the c.C248T mutation, which resulted in the emergence of genotype ee^a. Based on the association analysis of the horse MC1R genotype data (a total of 571 individuals) published by NCBI with their coat color traits, we found that there were two genotypes of EE and Ee in the bay and black horse populations, four genotypes of EE, Ee, ee and ee^ain gray horse population, and two genotypes of ee and ee^a in the chestnut horse population. The distribution of genotypes were extremely significantly different(P<0.01) in black and bay horse populations,showing Hardy-Weinberg disequilibrium, and the allele E was the dominant one. However, allele E was not detected in chestnut horses and e was the dominant allele. The result of phylogenetic tree showed that the individuals with EE genotype had the highest similarity to horse and Przewalskii. These results indicate that the different genotypes produced by the MC1R polymorphic loci are related to the bay,black,chestnut coat colors and the colorful hair color of horses. There was no ee genotype in bay and black horses and no allele E in chestnut horses. This study lay the foundation for the application of genotypes in the directional selection breeding of horse coat color.

中图分类号:

S821.2

赵珊珊, 肖宁, 李志鹏, 潘庆杰, 李桢, 石德顺, 刘庆友. 马MC1R基因多态位点与毛色相关性分析[J]. 畜牧兽医学报, 2018, 49(8): 1605-1616.

ZHAO Shan-shan, XIAO Ning, LI Zhi-peng, PAN Qing-jie, LI Zhen, SHI De-shun, LIU Qing-you. Correlation Analysis between the Polymorphism Loci of MC1R Gene and Coat Color in Horses[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1605-1616.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.08.005

https://www.xmsyxb.com/CN/Y2018/V49/I8/1605

参考文献

[1] 李蓓,何小龙,赵一萍,等.马毛色遗传的分子基础与应用[J].遗传,2010,32(11):1133-1140.
LI B,HE X L,ZHAO Y P,et al.Molecular basis and applicability in equine color genetics[J].Hereditas (Beijing),2010,32(11):1133-1140. (in Chinese)
[2] 杨永升,李宁,邓学梅,等.黑素皮质素受体1-哺乳动物黑色素形成中的关键基因[J].遗传,2004,26(4):544-550.
YANG Y S,LI N,DENG X M,et al.MC1R-the key gene in mammalian melanin synthesis[J].Hereditas (Beijing),2004,26(4):544-550. (in Chinese)
[3] 吴宇婷.哺乳动物毛色形成机制与影响因素[J].四川动物,2011,30(6):1003-1007.
WU Y T.The formation mechanism of mammal coat color and its possible influencing factors[J].Sichuan Journal of Zoology,2011,30(6):1003-1007. (in Chinese)
[4] 徐桂利,张文香,段玲欣,等.狐狸MC1R基因编码区c.40A>C和c.41C>T相邻变异研究[J].畜牧兽医学报,2016,47(10):2020-2026.
XU G L,ZHANG W X,DUAN L X,et al.Study on adjacent variation of c.40A>C and c.41C>T of fox MC1R gene in different populations[J].Acta Veterinaria et Zootechnica Sinica,2016,47(10):2020-2026. (in Chinese)
[5] 封竣淇,徐伟,黄兰,等.MC1R基因的研究进展[J].中国畜牧兽医,2017,44(4):1141-1148.
FENG J Q,XU W,HUANG L,et al.Researchadvances on MC1R gene[J].China Animal Husbandry & Veterinary Medicine,2017,44(4):1141-1148. (in Chinese)
[6] 孙笑尉,张雪莲,庞全海,等.RNAi对羊驼黑色素细胞中MC1R表达量及黑色素合成的影响[J].畜牧兽医学报,2015,46(7):1163-1168.
SUN X W,ZHANG X L,PANG Q H,et al.Effect of RNAi onMC1R expression and melanin synthesis in the melanocytes of alpaca[J].Acta Veterinaria et Zootechnica Sinica,2015,46(7):1163-1168. (in Chinese)
[7] 高荣琨,陈伟,曲海娥,等.黑素皮质素受体1(MC1R)基因与VC-獭兔毛色关系的研究[J].畜牧兽医学报,2013,44(6):888-893.
GAO R K,CHEN W,QU H E,et al.Study on the relationship between the melanocortin 1 receptor gene (MC1R) and the coat color in Rex-rabbits[J].Acta Veterinaria et Zootechnica Sinica,2013,44(6):888-893. (in Chinese)
[8] RIEDER S,TAOURIT S,MARIAT D,et al.Mutations in the agouti (ASIP),the extension (MC1R),and the brown (TYRP1) loci and their association to coat color phenotypes in horses (Equus caballus)[J].Mamm Genome,2001,12(6):450-455.
[9] WAGNER H J,REISSMANN M.New polymorphism detected in the horse MC1R gene[J].Anim Genet,2000,31(4):289-290.
[10] 李蓓,芒来.蒙古马黑素细胞激素受体1(MC1R)基因多态性与毛色表型相关性研究[J].黑龙江畜牧兽医,2011(5):43-45.
LI B,MANG L.Study on the correlation between mongolian horse melanocyte cytokine receptor 1(MC1R) gene polymorphism and coat phenotype[J].Heilongjiang Animal Science and Veterinary Medicine,2011(5):43-45. (in Chinese)
[11] LUDWIG A,PRUVOST M,REISSMANN M,et al.Coat color variation at the beginning of horse domestication[J].Science,2009,324(5926):485.
[12] KIM N Y,SHIN K Y,LEE C E,et al.Genetic characterization of Wolla coat color in Jeju horses[J].J Anim Sci Technol,2012,54(5):375-379.
[13] KAKOI H,TOZAKI T,NAGATA S,et al.Development of a method for simultaneously genotyping multiple horse coat colour loci and genetic investigation of basic colour variation in Thoroughbred and Misaki horses in Japan[J].J Anim Breed Genet,2009,126(6):425-431.
[14] 王超,朱建伟,张海仓,等.蛋白质三级结构预测算法综述[J].计算机学报,2016,39(24):1-21.
WANG C,ZHU J W,ZHANG H C,et al.A survey on algorithms for protein tertiary structure prediction[J].Chinese Journal of Computers,2016,39(24):1-21. (in Chinese)
[15] 乔辉,李晓琴,徐海松,等.阳离子-π相互作用在两种典型蛋白质折叠型中的偏好性[J].生物信息学,2011,9(1):76-81.
QIAO H,LI X Q,XU H S,et al.The preference of cation-π interactions in two typical protein folds[J].China Journal of Bioinformatics,2011,9(1):76-81. (in Chinese)
[16] THIRUVENKADAN A K,KANDASAMY N,PANNEERSELVAM S.Coat colour inheritance in horses[J].Livest Sci,2008,117(2-3):109-129.
[17] GETHER U.Uncovering molecular mechanisms involved in activation of G protein-coupled receptors[J].Endocr Rev,2000,21(1):90-113.
[18] NORDLUND J J,BOISSY R E,HEARING V J,et al.The pigmentary system:Physiology and pathophysiology[M].New York:Oxford University Press,1998:155-156.
[19] 党珍,王嘉福,赵星艳,等.黑色素皮质激素受体1基因248位点多态性与中国马品种毛色的相关性分析[J].中国畜牧兽医,2012,39(6):149-152.
DANG Z,WANG J F,ZHAO X Y,et al.Association of polymorphism inMC1R gene at site 248 with chestnut coat color in three Chinese horse breeds[J].China Animal Husbandry & Veterinary Medicine,2012,39(6):149-152. (in Chinese)
[20] BOX N F,WYETH J R,O'GORMAN L E,et al.Characterization of melanocyte stimulating hormone receptor variant alleles in twins with red hair[J].Hum Mol Genet,1997,6(11):1891-1897.
[21] SCHAFFER J V,BOLOGNIA J L.The melanocortin-1 receptor:Red hair and beyond[J].Arch Dermatol,2001, 137(11):1477-1485.
[22] FLANAGAN N,HEALY E,RAY A,et al.Pleiotropic effects of the melanocortin 1 receptor (MC1R) gene on human pigmentation[J].Hum Mol Genet,2000,9(17):2531-2537.
[23] HEALY E,JORDAN S A,BUDD P S,et al.Functional variation of MC1R alleles from red-haired individuals[J].Hum Mol Genet,2001,10(21):2397-2402.
[24] WOLF HORRELL E M,BOULANGER M C,D'ORAZIO J A.Melanocortin 1 receptor:Structure,function,and regulation[J].Front Genet,2016,7:95.
[25] DOAN R,COHEN N D,SAWYER J,et al.Whole-Genome sequencing and genetic variant analysis of a quarter horse mare[J].BMC Genomics,2012,13:78.
[26] 白东义,杨丽华,乌尼尔夫,等.Kit基因对白马被毛褪色的影响[J].遗传,2011,33(11):1171-1178.
BAI D Y,YANG L H,UNERHU,et al.Effects of Kit gene on coat depigmentation in white horses[J].Hereditas (Beijing),2011,33(11):1171-1178. (in Chinese)
[27] NEGRO S,IMSLAND F,VALERA M,et al.Association analysis of KIT,MITF,and PAX3 variants with white markings in Spanish horses[J].Anim Genet,2017,48(3):349-352.
[28] IMSLAND F,MCGOWAN K,RUBIN C J,et al.Regulatory mutations in TBX3 disrupt asymmetric hair pigmentation that underlies Dun camouflage color in horses[J].Nat Genet,2016,48(2):152-158.
[29] HAUSWIRTH R,HAASE B,BLATTER M,et al.Mutations in MITF and PAX3 cause "splashed white" and other white spotting phenotypes in horses[J].PLoS Genet,2012,8(4):e1002653.
[30] STACHURSKA A,PIETA M,LOJEK J,et al.Performance in racehorses of various colours[J].Livest Sci,2007, 106(2-3):282-286.
[31] STACHURSKA A,BRODACKI A.Variation of gene frequencies in ASIP,MC1R and GREY loci in Thoroughbred horses[J].Livest Sci,2008,113(2-3):163-168.
[32] JACOBS L N,STAIGE E A R,ALBRIGHT J D,et al.The MC1R and ASIP coat color loci may impact behavior in the horse[J].J Hered,2016,107(3):214-219.