中国荷斯坦牛CXCR1基因编码区SNP多态与临床乳房炎和生产寿命的关联分析

doi:10.3864/j.issn.0578-1752.2017.12.016

摘要/Abstract

摘要： 【目的】探讨中国荷斯坦牛CXCR1基因编码区SNP突变与临床乳房炎和生产寿命的相关性。【方法】根据CXCR1基因编码区序列，利用PCR-直接测序法对低SCS和高SCS样本各20个样本进行SNP筛查，最后对所选择的4个SNP位点利用飞行质谱法对866头中国荷斯坦牛进行检测，同时收集所检测牛只临床乳房炎和生产寿命等信息，利用多因素方差分析法、Logistic回归、Cox生存回归等方法分析以上SNP位点突变与临床乳房炎发生次数和生产寿命的相关性。【结果】CXCR1基因编码区共发现13个SNP位点（291 C＞T、333 C＞G、337 A＞G、365 C＞T、570 A＞G、642 A＞G、735 C＞G、816 A＞C、819 A＞G、980 A＞G、995 A＞G、1008 C＞T和1068 A＞G），分为4个连锁群，随后从每个连锁群中各选择1个SNP位点（642 A＞G, 816 A＞C, 980 A＞G和1068 A＞G），利用飞行质谱法对大样本中国荷斯坦牛进行检测。4个SNP位点共有9种单倍型，其中单倍型GAGG频率最高（0.3141），而单倍型ACAA频率最低（0.0017）。CXCR1-642与2胎牛患临床乳房炎次数有显著相关（P＜0.05），AG基因型个体2胎奶牛患临床乳房炎次数显著高于AA基因型（P＜0.05），CXCR1-816 AA 基因型个体3胎奶牛患临床乳房炎次数显著低于AC和CC基因型（P＜0.05），其它SNP位点与各胎次临床乳房炎发生次数均无显著相关（P＞0.05）。CXCR1-816与奶牛离群月龄有极显著相关（P＜0.01），与奶牛生产月龄和离群胎次有显著相关（P＜0.05），CXCR1-816 AA基因型个体生产月龄、离群月龄和离群胎次均显著高于CC基因型个体（P＜0.05），而其它SNP位点对生产月龄、离群月龄和离群胎次均无显著相关（P＞0.05）。Cox生存分析表明：只有CXCR1-816位点与奶牛生存时间有显著相关(P＜0.05)，CXCR1-816 CC基因型个体在各时间段的生存概率均低于AA和AC基因型个体。【结论】CXCR1-816 A＞C突变与中国荷斯坦牛患临床乳房炎次数和生产寿命有显著相关，在进一步验证其功能后，可用于中国荷斯坦牛生产寿命的分子标记辅助选择。

关键词: 中国荷斯坦牛, CXCR1, SNP, 临床乳房炎, 生产寿命

Abstract: 【Objective】 The objective of this study was to investigate the association between the SNPs in the coding sequence (CDS) of CXCR1 gene and the clinical mastitis and lifetime for Chinese Holstein.【Method】SNPs in the CDS of CXCR1 gene were screened by using PCR and direct sequencing for 20 cows with low SCS and 20 cows with high SCS. Finally, the selected SNPs of CXCR1 gene for 866 Chinese Holstein cows were detected using flight mass spectrometry. The clinical mastitis and productive life of tested cows were collected from the management system of dairy farm. The association between the SNPs and the clinical mastitis and productive life of tested cows was analyzed using multi factor variance analysis, Logistic regression, Cox regression.【Result】A total of 13 SNPs were found in the CDS region of CXCR1 gene (291 C＞T, 333 C＞G, 337 A＞G, 365 C＞T, 570 A＞G, 642 A＞G, 735 C＞G, 816 A＞C, 819 A＞G, 980 A＞G、, 995 A＞G, 1008 C＞T and 1068 A＞G), and these SNPs were divided into 4 linkage groups, and four SNPs were selected from each linkage group for further analysis（642 A＞G, 816 A＞C, 980A＞G and 1068 A＞G). A total of 9 haplotypes were observed for 4 SNPs of CXCR1 gene, the GAGG haplotype frequency was the highest (0.3141), and the frequency of haplotype ACAA was the lowest (0.0017). CXCR1-642 A＞G showed a significant association with the number of dairy cows suffering from clinical mastitis in second lactation (P＜0.05). The number of dairy cows suffering from clinical mastitis for individual with AG genotype was significantly higher than that of AA genotype (P＜0.05). The number of dairy cows suffering from clinical mastitis for individual with CXCR1-816 AA genotype was significantly higher than that of AC and CC genotypes (P＜0.05). The other SNPs showed no significant association with the number of clinical mastitis. CXCR1-816 A＞C mutation showed an extremely significant association with culling age (P＜0.01) and a significant association with productive month and culling lactation (P＜0.05). The productive month, culling age and culling lactation for the individual of CXCR1-816 AA were significantly higher than that of CC genotype (P＜0.05). Cox regression showed that only CXCR1-816 showed significant association with the productive life（P＜0.05), the survival probability of individual with CC genotype was significantly lower than that of AA and AC genotype in each time period. 【Conclusion】The mutation of CXCR1-816 A＞C showed significant association with the clinical mastitis and lifetime for Chinese Holstein, and this SNP could be used for molecular marker-assisted selection of productive life for Chinese Holstein when the molecular function of this mutation were proved.

Key words: Chinese Holstein, CXCR1, SNPs, clinical mastitis, productive lifetime

王梦琦，倪炜，张慧敏，杨章平，王西朴，蒋彦森，毛永江. 中国荷斯坦牛CXCR1基因编码区SNP多态与临床乳房炎和生产寿命的关联分析[J]. 中国农业科学, 2017, 50(12): 2359-2370.

WANG MengQi, NI Wei, ZHANG HuiMin, YANG ZhangPing, WANG XiPu, JIANG YanSen, MAO YongJiang. Association Between SNPs in the CDS Regions of CXCR1 Gene and the Clinical Mastitis and Lifetime for Chinese Holstein[J]. Scientia Agricultura Sinica, 2017, 50(12): 2359-2370.

0
/ / 推荐

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

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2017.12.016

https://www.chinaagrisci.com/CN/Y2017/V50/I12/2359

参考文献

[1] MIDDLETON J R, SAEMAN A, FOX L K, LOMBARD J, HOGAN J S, SMITH K L. The National mastitis council: A global organization for mastitis control and milk quality, 50 years and beyond. Journal of Mammary Gland Biology & Neoplasia, 2014, 19(3-4): 241-251.

[2] 何阳花. CD4 与STAT5b 基因单核苷酸多态性与SCC 及产奶性状的关联分析[D]. 北京: 中国农业大学, 2009.

HE Y H. Association analysis between the single nucleotide polymorphism of CD4 and STAT5b gene and SCC and milk traits[D]. Beijing: China Agricultural University, 2009.(in Chinese)

[3] 李芬，周娟，马云，张立岗. 奶牛生产寿命和产奶量的相关性研究. 中国牛业科学, 2006, 32(6):12-14.

LI F, ZHOU J, MA Y, ZHANG L G. Study of association between production life of dairy cattle and its milk yield. China Cattle Science, 2006, 32(6):12-14.(in Chinese)

[4] WOLFOVA M, WOLF J, KVAPILIK J, KICA J. Selection for profit in cattle: I. Economic weights for purebred dairy cattle in the Czech Republic. Journal of Dairy Science, 2007, 90: 2442-2455.

[5] KOECK A, LOKER S, MIGLIOR F, KELTON D F, JAMROZIK J, SCHENKEL F S. Genetic relationships of clinical mastitis, cystic ovaries, and lameness with milk yield and somatic cell score in first-lactation Canadian Holsteins. Journal of Dairy Science, 2014，97:5806-5813.

[6] ZHANG Q, GULDBRANDTSEN B, THOMASEN J R, LUND M S, SAHANA G. Genome-wide association study for longevity with whole-genome sequencing in 3 cattle breeds. Journal of Dairy Science, 2016, 99:7289-7298.

[7] HOCH R C, SCHRAUFSTAETTER I U, COCH RANE C G. In vivo, in vitro, and molecular aspects of interleukin-8 and interleukin-8 receptors. Journal of Laboratory & Clinical Medicine, 1996, 128(2): 134-145.

[8] WOLF M, DELGADO M B, JONES S A, DEWALD B, CLARK-LEWIS, BAQQIOLINI M. Granulocyte chemotactic protein 2 acts via both IL8 receptors, CXCR1 and CXCR2. European Journal of Immunology， 1998, 28(1): 164-170.

[9] PAAPE M, MEHRZAD J, ZHAO X, DETILLEUX J, BURCENICH C. Defense of the bovine mammary gland by polymorpho-nuclear neutrophil leukocytes. Journal of Mammary Gland Biology and Neoplasia, 2002, 7(2): 109-121.

[10] BAGGIOLINI M, DEWALD B, MOSER B. Interleukin-8 and related chemotactic cytokines-CXC and CC chemokines. Advances in Immunology, 1994, 55(55):97-179.

[11] BANNERMAN D D, PAAPE M J, LEE J W, ZHAO X, HOPE J C, RAINARD P. Escherichia coli and Staphylococcus aureus elicit differential innate immune responses following intramammary infection. Clinical & Diagnostic Laboratory Immunology, 2004, 11(3): 463-472.

[12] RIOLLET C, RAINARD P, POUTREL B. Differential induction of complement fragment C5a and inflammatory cytokines during intramammary infections with Escherichia coli and Staphylococcus aureus. Clinical and Diagnostic Laboratory Immunology, 2000, 7: 161-167.

[13] BARBER M R, YANG T J. Chemotactic activities in nonmastitic and mastitic mammary secretions: Presence of interleukin-8 in mastitic but not non-mastitic secretions. Clinical & Diagnostic Laboratory Immunology, 1998, 5(1): 82-86.

[14] LEYVA-BACA I, SCHENKEL F, SHARMA B S, JANSEN G B, AND KARROW N A. Identification of single nucleotide polymorphisms in the bovine CCL2, IL8, CCR2 and IL8RA genes and their association with health and production in Canadian Holsteins. Animal Genetics, 2007, 38(3):198-202.

[15] YOUNGERMAN S M, SAXTON A M AND PIGHETTI G M. Novel single nucleotide polymorphism and haplotypes within the bovine CXCR2 gene. Immunogenetics, 2004, 56: 355-359.

[16] YOUNGERMAN S M, SAXTON A M, OLIVER S P AND PIGHETTI G M. Association of CXCR2 polymorphisms with subclinical and clinical mastitis in dairy cattle. Journal of Dairy Science, 2004, 87: 2442-2448.

[17] 周雷，王洪梅，王长法，柳楠，侯明海，李秋玲，仲跻峰. 中国荷斯坦牛CXCR1基因第二外显子新SNPs与乳腺炎的关联分析. 畜牧兽医学报. 2011 , 42(8):1063-1070.

ZHOU L, WANG H M, WANG C F, LIU N, HOU M H, LI Q L，ZHONG J F. New genetic polymorphisms within Exon 2 of Chinese Holstein Cattle CXCR1 gene and its association with mastitis. Acta Veterinaria et Zootechnica Sinica, 2011, 42(8):1063-1070. (in Chinese)

[18] CHAPINAL N, BARRIENTOS A K, KEYSERLINGK M A G, GALO E, AND WEARY D M. Herd-level risk factors for lameness in free-stall farms in the northeastern United States and California. Journal of Dairy Science, 2012, 96(1):318-328.

[19] KHATIB H, HEIFETZ E, DEKKERS J C. Association of the protease inhibitor gene with production traits in Holstein dairy cattle. Journal of Dairy Science, 2005, 88(3):1208-1213.

[20] KHATIB H, SCHUTZKUS V, CHANG Y M, ROSA G J. Pattern of expression of the uterine milk protein gene and its association with productive life in dairy cattle. Journal of Dairy Science, 2007, 90(5):2427-2433.

[21] ASHWELL M S, REXROAD C E, MILLER R H, VANRADEN P M，DA Y. Detection of loci affecting milk production and health traits in an elite US Holstein population using microsatellite markers. Animal Genetics, 1997, 28(3): 216-222.

[22] ASHWELL M S, VAN TASSELL C P. Detection of putative loci affecting milk, health, and type traits in a US Holstein population using 70 microsatellite markers in a genome scan. Journal of Dairy Science, 1999, 82(11): 2497-2502.

[23] HEYEN D W,WELLER J I , RON M, BAND M, BEEVER J E, FELDMESSER E, DA Y, WIGGANS G R, VANRADEN P M, LEWIN H A. A genome scan for QTL influencing milk production and health traits in dairy cattle. Physiological Genomics, 1999, 1(3):165-175.

[24] KOMISAREK J, DORYNEK Z. Effect of ABCG2, PPARGC1A, OLR1 and SCD1 gene polymorphism on estimated breeding values for functional and production traits in Polish Holstein-Friesian bulls . Journal of Applied Genetics, 2009, 50: 125-132.

[25] COLE J B, GEORGE R, WIGGANS L M, SONSTEGARD T S, LAWLORJR T J, CROOKER B A, VAN TASSELL C P, YANG J, WANG S W, MATUKUMALLI L K, YANG D. Genome-wide association analysis of thirty one production, health, reproduction and body conformation traits in contemporary U. S. Holstein cows. BMC Genomics, 2011, 12(408): 1471-2164.

[26] CHEBEL R C, SUSCA F, SANTOS J E. Leptin genotype is associated with lactation performance and health of Holstein cows. Journal of Dairy Science, 2008, 91(7):2893-2900.

[27] 徐敏, 平富强, 陈仕毅, 赖松家, 刘益平. CXCR2 基因多态性与奶牛乳房炎和乳品质的关联. 遗传, 2008, 30: 463-468.

XU M, PING F Q, CHEN S Y, LAI S J, LIU Y P. Study on the relationships between polymorphisms of CXCR2 gene and milk quality and mastitis of dairy cow. Herditas, 2008, 30:463-468.(in Chinese)

[28] 官久强, 王洪梅, 王长法, 李秋玲, 李建斌, 帅素容, 侯明海, 仲跻峰. 中国荷斯坦牛白介素8 受体基因编码区多态性与乳腺炎的关联分析. 中国农业科学, 2010, 43(5): 1057-1065.

GUAN J Q, WANG H M, WANG C F, LI Q L, LI J B, SHUAI S R, HOU M H, ZHONG J F. Genetic Polymorphisms Within the Coding Regions of IL8R Gene and Its Association with Mastitis Trait in Chinese Holstein Cattle. Scientia Agricultura Sinica, 2010, 43(5): 1057-1065. (in Chinese)

[29] SHI Y Y, HE L. SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Research, 2005, 15(2):97-98.

[30] LEE J W, BANNERMAN D D, PAAPE M J, HUANG M K, ZHAO X. Characterization of cytokine expression in milk somatic cells during intramammary infections with Escherichia coli or Staphylococcus aureus by real-time PCR. Veterinary Research, 2006, 37(2):219-229.

[31] PIGHETTI G M, KOJIMA C J, WOJAKIEWICZ L, RAMBEAUD M. The bovine CXCR1 gene is highly polymorphic. Veterinary Immunology and Immunopathology, 2011, 145 (1-2): 464-470.

[32] 官久强, 王洪梅, 王长法, 李秋玲, 仲跻峰, 侯明海, 帅素容. 中国荷斯坦牛CXCR1基因编码区的遗传多态性. 西北农林科技大学学报(自然科学版), 2009, 37(6): 47-52.

GUAN J Q, WANG H M, WANG C F, LI Q L, ZHONG J F, HOU M H, SHUAI S R. Genetic polymorphisms within the coding regions of CXCR1 gene in Chinese Holstein cattle. Journal of Northwest A &F University (Nat. Sci. Ed .), 2009, 37(6): 47-52. (in Chinese)