Association Analysis of Polymorphism of KRTCAP3 Gene with Wool Production and Growth Performance in Fine Wool Sheep (Ovis aries)
WANG Li1, HU Rui-Xue2, LI Fa-Di1, YUE Xiang-Peng1,*
1 State Key Laboratory of Grassland Agro-ecosystems/Key Laboratory of Grassland Livestock Industry Innovation/Ministry of Agriculture and Rural Affairs/Engineering Research Center of Grassland Industry, Ministry of Education/College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730020, China; 2 College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
摘要角化细胞相关蛋白3基因(keratinocyte-associated protein 3, KRTCAP3)主要调节角蛋白表达进而影响机体皮肤、指甲和毛发的生长发育。为了解KRTCAP3基因在'南甘'杂交绵羊(Ovis aries)群体中的遗传变异及其与产毛和生长性状之间的关系,本研究以南非肉用美利奴(Ovis aries)(♂)×甘肃高山细毛羊(♀)('南甘')及南非肉用美利奴(♂) ×'南甘' (♀) ('南南甘')杂种羊为研究对象,采用DNA混合池测序结合SNPscanTM高通量SNP分型技术对KRTCAP3的基因全长进行SNP扫描,同时分析SNP位点与杂种羊产毛和生长性状的相关性。结果表明,在KRTCAP3基因第2内含子上发现1个g.34287851C>G的SNP位点,鉴定到CC、CG和GG 3种基因型,其基因型频率分别为0.35、0.52和0.13,C和G等位基因频率分别为0.61和0.39。遗传参数分析发现,杂合度(heterozygosity, He)、纯合度(homozygosity, Ho)、有效等位基因数(effective number of alleles, Ne)和多态信息含量(polymorphic information content, PIC)分别为0.47、0.53、1.90和0.36,属于中度多态(0.25<PIC<0.50)。χ2适合性检验表明,该SNP在研究群体中不处于Hardy-Weinberg平衡状态(P<0.05)。SNP与毛用性状的关联分析发现,该SNP位点与肩部和体侧毛长、股部和背部毛长、羊毛卷曲度和卷曲回复率相关(P<0.05或P<0.01);SNP与生长性状的关联分析发现,该位点与管围和胸围性状相关(P<0.05或P<0.01)。综上,KRTCAP3基因g.34287851C>G位点可作为分子标记辅助选育的候选分子标记,在细毛羊品种选育和培育肉毛兼用新品种中利用,加快育种进程。
Abstract：The keratinocyte-associated protein 3 gene (KRTCAP3) affects the growth of skin, nails and hair by regulating keratin. In order to uncover the genetic variation in the KRTCAP3 gene and its genetic roles in wool production and growth performance in nangan (South African Mutton Merino (♂)×Gansu alpinefine-wool (♀)) and nannangan (South African Mutton Merino (♂)×nangan (♀)) crossbred sheep breed. DNA pool sequencing was used to scan the potential SNPs within the full-length DNA of KRTCAP3 gene in AF×MF population. The correlation between SNP locus and hair growth and growth traits of hybrid sheep was also analyzed. Six hundred crossbred sheep were individually genotyped by SNPscanTM method. Then, the association analysis between SNP locus and wool production and growth performance was implemented. The results showed that one SNP (g.34287851C>G) was identified in intron2. Three genotypes, CC, CG and GG, in this locus were found in AF×MF crossbreed sheep population. The genotypic frequencies of CC, CG and GG were 0.35, 0.52 and 0.13, respectively. The allelic frequencies of C and G were 0.61 and 0.39, respectively. The population genetic parameters analyses showed that the homozygosity (Ho), heterozygosity (He) and effective allele number (Ne) were 0.53, 0.47 and 1.90, respectively. The polymorphism information contents (PIC) was 0.36 and suggested that this SNP was moderate diversity (0.25<PIC<0.5). The χ2 test indicated that this locus was not in Hardy-Weinberg equilibrium (P<0.05). The association analysis of polymorphisms in KRTCAP3 gene with wool traits revealed that g.34287851C>G was significantly associated with the wool length of shoulder and side , wool length of thigh and notum, crimpness and crimp recovery (P<0.05 or P<0.01). The association analysis of polymorphisms in KRTCAP3 gene with growth performance revealed that g.34287851C>G was significantly associated with tube circumference and chest girth (P<0.05 or P<0.01). In summary, the g.34287851C>G site in KRTCAP3 gene could be used as a useful molecular marker for selection wool traits and growth performance. This study provides a molecular basis for utilization of fine wool resources and accelerates the breeding progress.
王丽, 胡瑞雪, 李发弟, 乐祥鹏. KRTCAP3基因多态性与细毛羊产毛及生长性状的相关性分析[J]. 农业生物技术学报, 2020, 28(1): 84-91.
WANG Li, HU Rui-Xue, LI Fa-Di, YUE Xiang-Peng. Association Analysis of Polymorphism of KRTCAP3 Gene with Wool Production and Growth Performance in Fine Wool Sheep (Ovis aries). 农业生物技术学报, 2020, 28(1): 84-91.
[1] 常伟. 2013. 高寒牧区引入南非肉用美利奴与甘肃高山细毛羊的杂交效果观察[J].畜牧兽医杂志, 32(4): 64-65. (Chang W.2013. Observation on the hybridization effect of introducing south african meat merino and gansu alpine fine wool sheep in alpine pastoral area[J]. Journal of Animal Science and Veterinary Medicine, 32(4): 64-65.) [2] 陈宗芳. 2009. 浅析制约天祝草畜产业发展的原因及治理对策[J]. 中国牛业科学, 35(5): 58-61. (Chen Z F.2009. Brief analysis of the causes and managing measures of confining the development of grassland and aminal production[J]. China Cattle Science, 35(5): 58-61.) [3] 楚晓, 崔萍, 陈莉萍, 等. 2014. 南非肉用美利奴羊与甘肃高山细毛羊杂交一代羊毛品质分析[J]. 中国草食动物科学, 34(6): 13-14. (Chu X, Cui P, Chen L P, et al.2014. Quality analysis of hybrids of south african meat merino and gansu alpine fine wool sheep[J], Grass-Feeding Livestock, 34(6): 13-14.) [4] 胡瑞雪. 2018. 南甘杂种羊产毛和生长性能测定及其分子标记筛选[D]. 硕士学位论文, 兰州大学, 导师:李发弟, pp. 99-104. (Hu R X.2018. Wool production and growth performance measurement and related molecular markers screening of south african mutton merino×gansu alpine fine wool crossbred sheep[D]. Thesis for M.S., Lanzhou University, Supervisor:Li F D, pp. 99-104.) [5] 李锋红, 马友记, 常伟, 等. 2015. 南非肉用美利奴羊与甘肃高山细毛羊杂交羔羊屠宰性能和肉品质测定[J]. 中国草食动物科学, 35(3): 10-13. (Li F H, Ma Y J, Chang W, et al.2015. Analysis on slaughter performance and meat quality of F1 lambs between south african mutton merino and gansu alpine fine wool sheep[J], China Herbivores Science, 35(3): 10-13.) [6] 李锋红. 2018. 南非肉用美利奴与甘肃高山细毛羊杂交效果实验[J]. 中国畜禽种业, 4: 80-81. (Li F H.2018. Experiment on the hybrid effect of south african meat merino and gansu alpine fine wool sheep[J]. The Chinese Livestock and Poultry Breeding, 4: 80-81.) [7] 李桂英. 2017. 高山美利奴羊多胎、肉用品系选育方案[J]. 甘肃畜牧兽医, 47(4): 62-64. (Li G Y.2017. Alpine merino multi-child, meat product breeding program[J]. Gansu Animal Husbandry and Veterinary, 47(4): 62-64.) [8] 李娜, 盛迅伦. 2007. 基因内含子突变与视网膜色素变性的研究进展[J]. 国际眼科杂志, 7(1): 147-150. (Li N, Sheng X L.2007. Progress in the research of mutation of genetic intron and retinitis pigmentosa[J]. International Journal of Ophthalmology, 7(1): 147-150.) [9] 李岩. 2015. 导入南非美利奴对甘肃高山细毛羊F1代周岁公母羊生产性能的影响[J]. 甘肃畜牧兽医, 45(6): 41-43. (Li Y.2015. Effects of introduction of south african merino on the performance of F1 generation of male and female ewes in gansu[J]. Gansu Animal Husbandry and Veterinary, 45(6): 41-43.) [10] 王喜军. 2017. 甘肃高山细毛羊资源保护及利用研究[J]. 甘肃畜牧兽医, 47(4): 33-38. (Wang X J.2017. Study on the protection and utilization of alpine fine-wool sheep resources in gansu province[J]. Gansu Animal Husbandry and Veterinary, 47(4): 33-38.) [11] 王新基, 冉玉明, 马友记. 2016. 南非肉用美利奴羊与甘肃高山细毛羊杂交后代羊毛品质比较[J].中国草食动物科学, 36(04): 67-69. (Wang X J, Ran Y M, Ma Y J.2016. Comparison of wool quality between hybrids of south african meat merino and gansu alpine fine wool sheep[J]. China Herbivore Science, 36(04): 67-69.) [12] 王遵宝, 赵宗胜, 吴洪宾, 等. 2011. 绵羊皮肤源 EST-SSR 标记与羊毛性状的关联性分析[J]. 农业生物技术学报, 19(06): 1056-1062. (Wang Z B, Zhao Z S, Wu H B, et al.2011. Correlation analysis of ovine skin derived EST-SSR markers with wool traits[J]. Journal of Agricultural Biotechnology, 19(06): 1056-1062.) [13] 谢晓玲, 马思雨, 吴希, 等. 2018. 两种新的F8内含子突变导致剪接异常的机制研究[J]. 诊断学理论与实践, 17(01): 32-37. (Xie X L, Ma S Y, Wu X, et al.2018. Molecular pathogenesis of two novel splice site mutations of F8 in hemophilia A[J]. Journal of Diagnostics Concepts & Practice, 17(01): 32-37.) [14] 曾献存. 2010. 新疆绵羊遗传多样性及主要经济性状候选基因研究[D]. 博士学位论文, 石河子大学, 导师:贾斌. pp. 15-19. (Zeng X C.2010. Studies on the genetic diversity of xinjiang sheep breeds and candidate genes of major economic traits[D]. Thesis for Ph.D., Shihezi University, Supervisor: Jia B, pp. 15-19.) [15] 张恩宇, 罗玉柱, 李少斌, 等. 2015. 甘肃高山细毛羊及其杂种羊GH基因第3内含子多态性与生长性状的相关性[J]. 华北农学报, 30(05): 71-76. (Zhang E Y, Luo Y Z, Li S B, et al.2015. Correlation ship investigation of GH gene intron 3 polymorphism with growth traits in gansu alpine merino and crossbred sheep[J]. Acta Agriculturae Boreali-Sinica, 30(5): 71-76.) [16] 赵冰茹, 黄锡霞, 田可川, 等. 2016. 基于DNA池重测序技术研究细毛羊LAMB1基因的SNPs分析[J]. 农业生物技术学报, 24(12): 1872-1881. (Zhao B R, Huang X X, Tian K C, et al.2016. SNPs analysis of LAMB1 gene in fine-wool sheep (Ovis aries) based on the DNA pools and re-sequencing technologies[J]. Journal of Agricultural Biotechnology, 24(12): 1872-1881.) [17] Bonkobara M, Das A, Takao J, et al.2003. Identification of novel genes for secreted and membrane-anchored proteins in human keratinocytes[J]. British Journal of Dermatology, 148(4): 654-664. [18] Chen X, Li S, Yang Y, et al.2012. Genome-wide association study validation identifies novel loci for atherosclerotic cardiovascular disease[J]. Journal of Thrombosis and Haemostasis, 10(8): 1508-1514. [19] Gui H F, Liu F, Gen H Y, et al.2014. Characterization of the Lect2 gene and its associations with resistance to the big belly disease in asian seabass[J]. Fish & Shellfish Immunol, 37(1): 131-138. [20] Haridas V, Kim S O, Nishimura G, et al.2005. Avicinylation (thioesterification): A protein modification that can regulate the response to oxidative and nitrosative stress[J]. Proceedings of the National Academy of Sciences of the USA, 102(29): 10088-10093. [21] Howard T D, Whittaker P A, Zaiman A L, et al.2001. Identification and association of polymorphisms in the Interleukin-13 gene with asthma and atopy in a dutch population[J]. American Journal of Respiratory Cell and Molecular Biology, 25(3): 377-384. [22] Keele G R, Prokop J W, He H, et al.2018. Genetic fine-mapping and identification of candidate genes and variants for adiposity traits in outbred rats[J]. Obesity, 26(1): 213-222. [23] Köchl S, Niederstätter H, Parson W.2005. DNA extraction and quantitation of forensic samples using the phenol-chloroform method and real-time PCR[J]. Methods in Molecular Biology, 297: 13-30. [24] Millar D S, Martin H, Chuzhanova N A, et al.2010. Characterisation of a functional intronic polymorphism in the human growth hormone (Gh1) gene[J]. Human Genomics, 4(5): 289-301. [25] Thomas G H, Ye Y, Ting J C, et al.2006. Analysis and visualization of chromosomal abnormalities in SNP data with SNPscan[J]. BMC Bioinformatics, 7(1): 25-45. [26] Wang Z, Zhang H, Yang H, et al.2014. Genome-wide association study for wool production traits in a chinese merino sheep population[J]. PLOS ONE, 9(9): 1-8. [27] Warrick E, Duval C, Nouveau S, et al.2017. Morphological and molecular characterization of actinic lentigos reveals alterations of the dermal extracellular matrix[J]. British Journal of Dermatology, 177(6): 1619-1632.
