牦牛乳铁蛋白基因多态性及其与乳品质性状的关联分析

doi:10.16431/j.cnki.1671-7236.2021.05.016

摘要/Abstract

摘要： 试验旨在检测牦牛乳铁蛋白（lactoferrin，LF）基因多态性，评估基因突变对牦牛乳品质性状的影响，以期丰富牦牛重要经济性状的分子遗传研究。应用PCR-SSCP方法，检测甘南牦牛、西藏牦牛和天祝白牦牛LF基因5'UTR和内含子4突变，结合甘南牦牛乳品质测定，评估基因突变对其乳品质性状的影响。结果表明，牦牛LF基因5'UTR检测到c.-568 G>A的突变，等位基因A₁和基因型A₁B₁频率最高，为优势等位基因和基因型；内含子4检测到c.499+56 C>A突变，基因型A₂B₂频率最高，为优势基因型，等位基因A₂为甘南牦牛和天祝白牦牛优势等位基因，而B₂为西藏牦牛优势等位基因；各类群牦牛检测区域多态信息含量（PIC）介于0.25~0.50，属中度多态。关联分析表明，甘南牦牛LF基因突变影响部分胎次甘南牦牛乳品质性状，其中5'UTR第2胎基因型A₁A₁和A₁B₁个体无脂固体物质百分含量显著高于B₁B₁个体（P<0.05）；内含子4第3胎基因型A₂B₂和B₂B₂个体乳脂率显著高于A₂A₂型个体（P<0.05），第4胎基因型B₂B₂个体的乳脂率显著低于A₂A₂和A₂B₂型个体（P<0.05）。以上结果表明，甘南牦牛、西藏牦牛和天祝白牦牛LF基因存在多态性，甘南牦牛LF基因5'UTR、内含子4突变分别影响无脂固体物质含量和乳脂率，可以作为潜在的分子遗传标记。

关键词: 牦牛; LF基因; PCR-SSCP; 乳品质性状

Abstract: In this study,polymorphisms of yak lactoferrin (LF) gene were investigated and their effects on some milk quality traits were estimated so as to enrich the molecular genetic data for yak.Variations in 5'UTR and intron 4 of LF gene were checked by PCR-SSCP in 3 yak populations (Gannan yak,Tianzhu White yak and Tibetan yak) and their effects on the milk quality traits were analyzed in Gannan yak.The results showed that:Variation of c.-568 G>A and c.499+56 C>A were found in 5'UTR and intron 4 of yak LF gene,respectively.Allele A₁,genotype A₁B₁ of 5'UTR and genotype A₂B₂ of intron 4 were the dominant allele and genotype in three yaks.The allele A₂ of intron 4 was dominant in Gannan yak and Tianzhu White yak,but allele B₂ was dominant in Tibetan yak.The polymorphism information content (PIC) in 5'UTR and intron 4 of LF gene in yaks ranged from 0.25 to 0.50,belonging to moderate polymorphism.The association analysis showed that the variations in LF gene affected the milk quality traits in different parities of Gannan yak.Individuals with genotypes A₁A₁ and A₁B₁ of 5'UTR in second parity yak had significantly higher non-fat solids rate than B₁B₁ genotype (P<0.05),the genotypes A₂B₂ and B₂B₂ of intron 4 in the third parity had significantly higher milk fat rate than A₂A₂ genotype (P<0.05),as well as yaks with B₂B₂ genotype in fourth parity had significant lower milk rate than A₂A₂ and A₂B₂ genotype (P<0.05).In conclusion,the polymorphisms were found in LF gene of Gannan yak,Tianzhu White yak and Tibetan yak,and these variations in 5'UTR and intron 4 that were association with content of non-fat solids and milk fat rate of Gannan yak,respectively,could be used as potential molecular genetic markers for milk quality traits.

Key words: yak; LF gene; PCR-SSCP; milk quality traits

中图分类号:

S823.8⁺5

杨晓斌, 何莉莉, 张扬, 周瑞锋, 后康丽, 张芸芸, 王月丽, 李青青, 石斌刚, 胡江. 牦牛乳铁蛋白基因多态性及其与乳品质性状的关联分析[J]. 中国畜牧兽医, 2021, 48(5): 1655-1663.

YANG Xiaobin, HE Lili, ZHANG Yang, ZHOU Ruifeng, HOU Kangli, ZHANG Yunyun, WANG Yueli, LI Qingqing, SHI Bingang, HU Jiang. Polymorphisms of LF Gene and Their Association with Milk Quality Traits in Yak[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1655-1663.

参考文献

[1] 周小玲,刘书杰,郝力壮,等.补饲油菜籽对放牧牦牛乳生产的影响[J].黑龙江畜牧兽医,2010,1:64-65. ZHOU X L,LIU S J,HAO L Z,et al.Effects of supplementary feeding of rapeseed on milk production of grazing yak[J].Heilongjiang Animal Science and Veterinary Medicine,2010,1:64-65.(in Chinese)
[2] 张容昶,胡江.牦牛生产技术[M].北京:金盾出版社,2002. ZHANG R C,HU J.Yak Production Technology[M].Beijing:Golden Shield Press,2002.(in Chinese)
[3] 占今舜,张彬,胡金杰.乳铁蛋白的生物学功能及在仔猪生产上的应用[J].养猪,2011,5:21-23. ZHAN J S,ZHANG B,HU J J.Biological function of lactoferrin and its application in piglet production[J].Swine Production,2011,5:21-23.(in Chinese)
[4] LACASSE P,LAUZON K,DIARRA M S,et al. Utilization of lactoferrin to fight antibiotic-resistant mammary gland pathogens[J].Journal of Animal Science,2008,86(13):66-71.
[5] VALENTI P,ANTONINI G.Lactoferrin:An important host defence against microbial and viral attack[J].Cellular and Molecular Life Sciences,2005,62(22):2576-2587.
[6] KAMINSKI S,OLESKI K,BRYM P,et al.Single nucleotide polymorphism in the promoter region of the lactoferrin gene and its associations with milk performance traits in Polish Holstein Friesian cows[J].Russian Journal of Genetics,2006,42(8):924-927.
[7] CARVAJAL A M,HUIRCAN P,LEPORI A.Single nucleotide polymorphisms in immunity-related genes and their association with mastitis in Chilean dairy cattle[J].Genetics and Molecular Research,2013,12(3):2702-2711.
[8] MUHAGHEGH-DOLATABADY M,SHAFAEIPOUR A.Single strand conformation polymorphisms(SSCPs) in the 5'-flanking region of lactoferrin gene and its association with milk production traits and somatic cell score in Holstein cattle[J].International Journal of Advanced Biological and Biomedical Research,2014,2(2):279-285.
[9] 李强子,马生虎,徐军,等.中国荷斯坦牛LF基因多态性与泌乳性状及乳腺炎的关联分析[J].浙江农业学报,2016,28(7):1142-1147. LI Q Z,MA S H,XU J,et al.Genetic polymorphism of LF gene and its associations with milk traits and mastitis of Chinese Holstein cattle[J].ACTA Agriculturae Zhejiangensis,2016,28(7):1142-1147.(in Chinese)
[10] 蒋艳艳,石斌刚,陈彦丽,等.牦牛LPL基因组织表达及其突变与乳质性状的关联分析[J].西北农林科技大学学报(自然科学版),2019,47(12):1-9. JIANG Y Y,SHI B G,CHEN Y L.Tissue expression and polymorphisms of LPL gene and their association with milk quality traits in Yak[J].Journal of Northwest A&F University (Natural Science Edition).2019,47(12):1-9.(in Chinese)
[11] HU J,ZHOU H,SYMTH A,et al.Polymorphism of the bovine ADRB3 gene[J].Molecular Biology Reports,2010,37(7):3389-3392.
[12] PATEL V,BHUSHAN B,KUMAR V,et al.Molecular characterization of lactoferrin gene and its association with mastitis in crossbred cattle[J].Indian Journal of Animal Research,2015,49(6):757-760.
[13] 周秀敏,李强子,毕英杰,等.荷斯坦牛LF基因5'UTR区多态性分析[J].中国奶牛,2017,3:13-16. ZHOU X M,LI Q Z,BI Y J,et al.Polymorphism analysis of 5'UTR region of LF gene in Holstein cattle[J].Chinese Dairy Cows,2017,3:13-16.(in Chinese)
[14] 李国华,张沅,李宁.荷斯坦牛乳铁蛋白基因部分序列的PCR-SSCP分析[J].农业生物技术学报,2001,9(2):139-141. LI G H,ZHANG Y,LI N.PCR-SSCP analysis of the partial sequence of Holstein bovine lactoferrin[J].Journal of Agricultural Biotechnology,2001,9(2):139-141.(in Chinese)
[15] O'HALLORAN F,BERRY D P,BAHAR B,et al.Polymorphisms in the bovine lactoferrin promoter are associated with reproductive performance and somatic cell count[J].Journal of Dairy Science,2010,93(3):1253-1259.
[16] 王洪梅,孔振兴,王长法,等.奶牛乳铁蛋白基因5'侧翼区遗传多态性及其与乳腺炎关联性分析[J].遗传,2009,31(4):393-399. WANG H M,KONG Z X,WANG C F,et al.Genetic polymorphism in 5'-flanking region of the lactoferrin gene and its associations with mastitis in Chinese Holstein cows[J].Hereditas,2009,31(4):393-399.(in Chinese)
[17] ZHENG J,ATHER J L,SONSTEGARD T S,et al.Characterization of the infection-responsive bovine lactoferrin promoter[J].Gene,2005,353(1):107-117.
[18] GUO B L,JIAO Y,HE C,et al.A novel polymorphism of the lactoferrin gene and its association with milk composition and body traits in dairy goats[J].Genetics and Molecular Research,2010,9(4):2199-2206.
[19] BELLAMY W,TAKASE M,YAMAUCHI K,et al.Identification of the bactericidal domain of lacto-ferrin[J].Biochimica et Biophysica Acta,1992,1121(1-2):130-136.
[20] PAWLIK A,SENDER G,SOBCZYNSKA M,et al.Association between lactoferrin single nucleotide polymorphisms and milk production traits in Polish Holstein cattle[J].Archiv Fur Tierzucht,2014,54(1):1-12.
[21] ARNOULD V M,SOYEURT H,GENGLER N,et al.Genetic analysis of lactoferrin content in bovine milk[J].Journal of Dairy Science,2009,92(5):2151-2158.
[22] 王梦琦,倪炜,唐程,等.中国荷斯坦牛乳铁蛋白启动子区-131C>T和-28A>C位点SNP与泌乳性状、乳房炎和生产寿命的关联分析[J].农业生物技术学报,2018,26(5):811-818. WANG M Q,NI W,TANG C,et al.Association analysis on the SNP of LF-131C>T and LF-28A>C with milk performance,clinical mastitis and lifetime for Chinese Holstein (Bos taurus)[J].Journal of Agricultural Biotechnology,2018,26(5):811-818.(in Chinese)
[23] 何高明,李大全,孙庆华.IL-8受体和乳铁蛋白基因多态性与奶牛隐性乳房炎关系的初步研究[J].华北农学报,2009,24(S1):251-254. HE G M,LI D Q,SUN Q H.Preliminary study of relationship between the IL-8 receptor and lactoferrin gene polymorphisms and subclinical mastitics dairy cows[J].Acta Agriculture Boreali-Sinica,2009,24(S1):251-254.(in Chinese)