内蒙古绒山羊KAP9.2基因遗传多态性及其与生长性状的关联分析

doi:10.16431/j.cnki.1671-7236.2022.11.017

中国畜牧兽医 ›› 2022, Vol. 49 ›› Issue (11): 4270-4278.doi: 10.16431/j.cnki.1671-7236.2022.11.017

内蒙古绒山羊KAP9.2基因遗传多态性及其与生长性状的关联分析

王聪亮¹, 付琪¹, 巨敏莹⁵, 白晶晶¹, 李新曙⁵, 宋晓越¹, 李陇平¹, 蓝贤勇², 屈雷¹, 张磊¹, 朱海鲸^1,3,4

1. 榆林学院, 陕西省绒山羊工程技术研究中心, 榆林 719000;
2. 西北农林科技大学动物科技学院, 杨凌 712100;
3. 陕西省"四主体一联合"肉羊工程技术校企联合研究中心, 神木 719318;
4. 陕西浩丽绒山羊科技发展有限公司, 子洲 718499;
5. 榆林市畜牧兽医服务中心, 榆林 719000

收稿日期:2022-03-26 出版日期:2022-11-05 发布日期:2022-11-04
通讯作者: 张磊, 朱海鲸 E-mail:nmgzhangl@126.com;haijing12585@163.com
作者简介:王聪亮,E-mail:wcl15596064203@126.com。
基金资助:
国家自然科学基金项目（31702115）；国家现代农业产业园建设绒山羊产业重点项目（2020HX02）；陕西省科技统筹创新工程计划项目（2014KTDZ02-01）；榆林学院博士科研启动基金项目（22GK24）

The Genetic Polymorphisms of KAP9.2 Gene and Its Correlation Analysis with Growth Traits of Inner Mongolia Cashmere Goats

WANG Congliang¹, FU Qi¹, JU Minying⁵, BAI Jingjing¹, LI Xinshu⁵, SONG Xiaoyue¹, LI Longping¹, LAN Xianyong², QU Lei¹, ZHANG Lei¹, ZHU Haijing^1,3,4

1. Shaanxi Province Engineering & Technology Research Center of Cashmere Goat, Yulin University, Yulin 719000, China;
2. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China;
3. Shaanxi Province ‘Four Subjects and One Joint’ Sheep and Goat Engineering & Technology University & Enterprise Alliance Research Center, Shenmu 719318, China;
4. Shaanxi Haoli Cashmere Goat Technology Development Co., Ltd., Zizhou 718499, China;
5. Yulin Animal Husbandry and Veterinary Service Center, Yulin 719000, China

Received:2022-03-26 Online:2022-11-05 Published:2022-11-04

摘要/Abstract

摘要： 【目的】探究内蒙古绒山羊KAP基因家族角蛋白相关蛋白9.2（KAP9.2）基因插入/缺失（insertion/deletion，InDel）突变及其与生长性状的关系，以期为内蒙古绒山羊分子育种提供理论基础。【方法】选取606只雌性阿拉善型内蒙古绒山羊为研究对象，采集耳组织提取DNA并测定其体重及体高、体长和胸围等体尺数据，利用琼脂糖凝胶电泳方法与直接测序技术检测KAP9.2基因的InDel突变，检测遗传多样性指标，利用SPSS 23.0软件分析突变位点与生长性状的关联性。【结果】在内蒙古绒山羊KAP9.2基因外显子区存在30 bp的InDel突变，产生II、ID和DD 3种基因型。多态信息含量（PIC）显示，该位点在育成羊群体中属于中度多态（0.25<PIC<0.50），在成年羊群体中属于低度多态（PIC<0.25），且均处于哈代-温伯格平衡状态（P>0.05）；在整个群体中，属于低度多态（PIC<0.25），但偏离哈代-温伯格平衡状态（P<0.05）。关联分析结果表明，在育成羊群体中，此30 bp的InDel突变对体长和十字部高有显著影响（P<0.05），对体高、胸深和胸宽有极显著影响（P<0.01）；在成年羊群体中，此突变对管围、胸深和胸宽有极显著影响（P<0.01）。进一步的分析发现，在育成羊和成年羊全部群体中，此突变对体长和十字部高有显著影响（P<0.05），对体高、胸深和胸宽有极显著影响（P<0.01）。【结论】 KAP9.2基因InDel突变对阿拉善型内蒙古绒山羊多个生长性状有显著或极显著影响，可作为内蒙古绒山羊优良性状选育的分子标记。

关键词: KAP9.2基因; 内蒙古绒山羊; 插入/缺失(InDel); 生长性状

Abstract: 【Objective】 The purpose of this study was to explore the relationship between insertion/deletion (InDels) mutation of KAP gene family keratin-associated protein 9.2 (KAP9.2) gene and growth traits of Inner Mongolia cashmere goat, in order to provide a theoretical basis for molecular breeding of goats.【Method】 A total of 606 female Inner Mongolia cashmere goats were selected as the research objects, the ear tissue was collected to extract DNA, and their body weight and body size, such as body height, length and chest circumference were measured.The InDel mutation of KAP9.2 gene was detected by agarose gel electrophoresis and direct sequencing technology, and genetic diversity indicators were detected.The association between mutation locus and growth traits was analyzed by SPSS 23.0 software.【Result】 There was a 30 bp InDelmutation in the exon region of the KAP9.2 gene in Inner Mongolia cashmere goats, which produced three genotypes of II, ID and DD.The polymorphism information content (PIC) showed that the locus was moderately polymorphic (0.25<PIC<0.50) in the breeding goat population, low polymorphism (PIC<0.25) in the adult goat population, and both were in Hardy-Weinberg equilibrium (P>0.05).In the whole population, the locus was low polymorphism (PIC<0.25), but deviated from Hardy-Weinberg equilibrium (P<0.05).The results of association analysis showed that in the breeding goat population, the 30 bp InDel mutation had significant effect on body length and height at hip cross (P<0.05), and had extremely significant effect on body height, chest depth and chest width (P<0.01).In the adult goat population, this mutation had extremely significant effect on cannon circumference, chest depth and chest width (P<0.01).Further analysis found that this mutation had significant effect on body length and height at hip cross (P<0.05), and had extremely significant effect on body height, chest depth and chest width (P<0.01) in the whole population.【Conclusion】 The InDel mutation of KAP9.2 gene had significant or extremely significant effect on many growth traits of Alashan-type Inner Mongolia cashmere goat, and could be used as a molecular marker for the breeding of excellent traits of Inner Mongolia cashmere goat.

Key words: KAP9.2 gene; Inner Mongolia cashmere goats; insertion/deletion (InDel); growth traits

中图分类号:

S823.8

王聪亮, 付琪, 巨敏莹, 白晶晶, 李新曙, 宋晓越, 李陇平, 蓝贤勇, 屈雷, 张磊, 朱海鲸. 内蒙古绒山羊KAP9.2基因遗传多态性及其与生长性状的关联分析[J]. 中国畜牧兽医, 2022, 49(11): 4270-4278.

WANG Congliang, FU Qi, JU Minying, BAI Jingjing, LI Xinshu, SONG Xiaoyue, LI Longping, LAN Xianyong, QU Lei, ZHANG Lei, ZHU Haijing. The Genetic Polymorphisms of KAP9.2 Gene and Its Correlation Analysis with Growth Traits of Inner Mongolia Cashmere Goats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4270-4278.

参考文献

[1] 王岩, 秦志宏, 丁伟东.内蒙古羊绒产业开发研究[J].内蒙古大学学报(人文社会科学版), 2001, 1:14-20. WANG Y, QIN Z H, DING W D.A study on the development of cashmere industry in Inner Mongolia[J].Journal of Inner Mongolia University(Philosophy and Social Sciences), 2001, 1:14-20.(in Chinese)
[2] 朱乐乐, 李保荣, 王军庆, 等.山羊绒纤维结构与性能研究现状[J].纺织科技进展, 2020, 10:15-18. ZHU L L, LI B R, WANG J Q, et al.Research status of cashmere fiber structure and properties[J]. Progress in Textile Science & Technology, 2020, 10:15-18.(in Chinese)
[3] 李学武, 刘燕, 王瑞军, 等.内蒙古绒山羊不同毛被类型产绒量和体重的遗传参数估计[J].中国农业科学, 2018, 51(12):2410-2417. LI X W, LIU Y, WANG R J, et al.Genetic parameter estimation of cashmere yield body weight at different staple types of Inner Mongolia cashmere goats[J]. Scientia Agricultura Sinica, 2018, 51(12):2410-2417.(in Chinese)
[4] 海龙, 阿娜尔, 苏雅, 等.内蒙古绒山羊羊绒纤维平均直径现状与分析[J].中国畜牧杂志, 2017, 53(10):126-130. HAI L, ANAER, SU Y, et al.Analysis of the average fiber diameter in Inner Mongolia White cashmere goats[J].Chinese Journal of Animal Science, 2017, 53(10):126-130.(in Chinese)
[5] LIU H, YUE C W, ZHANG W, et al.Association of the KAP8.1 gene polymorphisms with fibre traits in Inner Mongolian cashmere goats[J].Asian Australasian Journal of Animal Sciences, 2011, 24(10), 1341-1347.
[6] WANG J Q, CHE L J, HICKFORD J G H, et al.Identification of the caprine Keratin-Associated protein 20-2(KAP20-2) gene and its effect on cashmere traits[J].Genes, 2017, 8(11):328.
[7] COCKETT N E, SHAY T L, SMIT M.Analysis of the sheep genome[J].Physiological Genomics, 2001, 7(2):69-78.
[8] MATSUNAGE R, ABE R, ISHII D, et al.Bidirectional binding property of high glycine-tyrosine keratin-associated protein contributes to the mechanical strength and shape of hair[J].Journal of Structural Biology, 2013, 183(3):484-494.
[9] POWELL B C, ROGERS G E.The role of keratin proteins and their genes in the growth, structure and properties of hair[J].Experientia Supplementum, 1997, 78:59-148.
[10] GONG H, ZHOU H, FORREST R H, et al.Wool Keratin-associated protein genes in sheep——A review[J].Genes, 2016, 7(6):24.
[11] POWELL B C, LWIGH M J, WARD K A, et al.Mammalian keratin gene families:Organisation of genes coding for the B2 high-sulphur proteins of sheep wool[J].Nucleic Acids Research, 1983, 11(16):5327-5346.
[12] LIU H Y, LI N, JIA C L, et al.Effect of the polymorphisms of keratin associated protein 8.2 gene on fibre traits in Inner Mongolia cashmere goats[J].Asian Australasian Journal of Animal Sciences, 2007, 20(6):821-826.
[13] ZHAO M L, ZHOU H T, HICKFORD J G H, et al.Variation in the caprine keratin-associated protein 15-1(KAP15-1) gene affects cashmere fibre diameter[J].Archives Animal Breeding, 2019, 62(1):125-133.
[14] WANG J Q, HAO Z Y, ZHOU H T, et al.A keratin-associated protein (KAP) gene that is associated with variation in cashmere goat fleece weight[J].Small Ruminant Research, 2018, 167:104-109.
[15] WANG X, ZHAO Z D, XU H R, et al.Variation and expression of KAP9.2 gene affecting cashmere trait in goats[J].Molecular Biology Reports, 2012, 39(12):10525-10529.
[16] YU H, WANG X, CHEN H, et al.The polymorphism of a novel 30 bp-deletion mutation at KAP9.2 locus in the cashmere goat[J].Small Ruminant Research, 2008, 80(1-3):111-115.
[17] HE L B, KANG Z H, KANG Y X, et al.Goat CMTM2:mRNA expression profiles of different alternative spliced variants and associations analyses with growth traits[J]. 3 Biotech, 2020, 10(3):131.
[18] AIJANABI S M, MARTINEZ I.Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques[J].Nucleic Acids Research, 1997, 25(22):4692-4693.
[19] WANG Z Y, WANG Z X, LIU Y, et al.Genetic evaluation of fiber length and fiber diameter from Inner Mongolia White cashmere goats at different ages[J].Small Ruminant Research, 2015, 123(1):22-26.
[20] 杨洁, 赫佳, 王丹碧, 等.InDel标记的研究和应用进展[J].生物多样性, 2016, 24(2):237-243. YANG J, HE J, WANG D B, et al.Progress in research and application of InDel markers[J]. Biodiversity Science, 2016, 24(2):237-243.(in Chinese)
[21] 孙珂欣, 郭应威, 曹睿, 等.InDel标记在畜禽育种中的应用现状分析[J].家畜生态学报, 2020, 41(3):76-80. SUN K X, GUO Y W, CAO R, et al.Analysis on application status on InDel marking in livestock and poultry breeding[J]. Journal of Domestic Animal Ecology, 2020, 41(3):76-80.(in Chinese)
[22] ROGERS M A, LANGBEIN L, WINTER H, et al.Characterization of a cluster of Human high/ultrahigh sulfur keratin-associated protein genes embedded in the type Ⅰ keratin gene domain on chromosome 17q12-21[J]. Journal of Biological Chemistry, 2001, 276(22):19440-19451.
[23] ITENGE-MWEZA T O, FORREST R H, MCKENZIE G W, et al.Polymorphism of the KAP1.1, KAP1.3 and K33 genes in Merino sheep[J].Molecular and Cellular Probes, 2007, 21(5-6):338-342.
[24] 赵志东, 徐华荣, 屈雷, 等.羊绒生长期、休止期陕北白绒山羊皮肤组织中KAP9.2和Hoxc13基因的表达[J].西北农林科技大学学报(自然科学版), 2012, 40(9):24-28. ZHAO Z D, XU H R, QU L, et al.Expression analysis of KAP9.2 and Hoxc13 gene in skin tissues during anagen of Shaanbei White cashmere goat[J]. Journal of Northwest A & F University(Natural Science Edition), 2012, 40(9):24-28.(in Chinese)
[25] 孙凯, 赵孟丽, 郝志云, 等.陇东绒山羊KRTAP2-1基因鉴定及其对羊绒性状影响的研究[J].核农学报, 2021, 35(5):1091-1098. SUN K, ZHAO M L, HAO Z Y, et al.Identification of the KRTAP2-1 gene and Its effect on Longdong cashmere traits[J].Journal of Nuclear Agricultural Sciences, 2021, 35(5):1091-1098.(in Chinese)
[26] PARSONS Y M, COOPER D W, PIPER L R.Evidence of linkage between high-glycine-tyrosine keratin gene loci and wool fibre diameter in a Merino half-sib family[J].Animal Genetics, 2009, 25(2):105-108.
[27] GONG H, ZHOU H, DYER J M, et al.Identification of the keratin-associated protein 13-3(KAP13-3) gene in Sheep[J].Open Journal of Genetics, 2011, 1(3):60-64.
[28] 李志刚, 王丽, 李树伟.新疆南疆和田羊、卡拉库尔羊毛囊KAP7基因的原核表达及多克隆抗体制备[J].中国畜牧兽医, 2017, 44(12):3410-3417. LI Z G, WANG L, LI S W.Prokaryotic expression and preparation antibody of wool follicle KAP7 gene of Hetian sheep and Karakul sheep in southern Xinjiang[J].China Animal Husbandry & Veterinary Medicine, 2017, 44(12):3410-3417.(in Chinese)
[29] 吴庭辉, 李涛, 李树伟.平原型和田羊毛囊KAP4.2基因的克隆与原核表达[J].黑龙江畜牧兽医, 2019, 13:11-20. WU T H, LI T, LI S W.Cloning and prokaryotic expression of folliculus Pili KAP4.2 gene of Plain Hetian sheep[J].Heilongjiang Animal Science and Veterinary Medicine, 2019, 13:11-20.(in Chinese)
[30] 李梦云, 郑萍, 李婉涛, 等.PRKAG3基因在不同品种猪不同生长阶段骨骼肌中的表达差异及其表达量与肉质的关系[J].动物营养学报, 2017, 29(5):1661-1669. LI M Y, ZHENG P, LI W T, et al.The expression differential of PRKAG3 gene in muscle at different growth stages for different breeds of Pigs and the relationship between PRKAG3 gene expression level and meat quality[J].Chinese Journal of Animal Nutrition, 2017, 29(5):1661-1669.(in Chinese)
[31] 杨珂伟, 胡亮, 罗斌, 等.藏绵羊KAP3.2基因的cDNA克隆、序列分析及组织表达的研究[J].西南民族大学学报(自然科学版), 2014, 40(6):809-813. YANG K W, HU L, LUO B, et al.Cloning and tissue expression of KAP3.2 gene in Tibetan sheep[J].Journal of Southwest Minzu University(Natural Science Edition), 2014, 40(6):809-813.(in Chinese)
[32] AIELLO D, PATEL K, LASAGNA E.The myostatin gene:An overview of mechanisms of action and its relevance to livestock animals[J].Animal Genetics, 2018, 49(6):505-519.
[33] 胡馨予, 姚逸安, 胡情情, 等.羊毛角蛋白基因家族及其启动子调控作用研究进展[J].中国细胞生物学学报, 2021, 43(8):1705-1713. HU X Y, YAO Y A, HU Q Q, et al.Research progress of wool Keratin gene family and its promoter regulation role[J].Chinese Journal of Cell Biology, 2021, 43(8):1705-1713.(in Chinese)
[34] 马晓萌, 轩俊丽, 王慧华, 等.TXNRD1基因多态性与乌珠穆沁绵羊生长性状的关联分析[J].畜牧兽医学报, 2016, 47(5):909-921. MA X M, XUAN J L, WANG H H, et al.TXNRD1 gene polymorphisms and its association with growth traits in Ujumqin sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(5):909-921.(in Chinese)
[35] 艾锦新, 龙安炬, 罗卫星, 等.黔北麻羊FABP3基因多态性及其与生长性状的相关性研究[J].畜牧与兽医, 2021, 53(1):13-18. AI J X, LONG A J, LUO W X, et al.Polymorphism of the FABP3 gene and its correlation with growth traits in Qianbei Ma goat[J].Animal Husbandry & Veterinary Medicine, 2021, 53(1):13-18.(in Chinese)
[36] 吴玉江, 富国文, 益西多吉, 等. KAP6.3基因与西藏绒山羊生产性状关系的研究[J].家畜生态学报, 2014, 35(12):16-20. WU Y J, FU G W, YIXI D J, et al.Correlation between KAP6.3 gene and production traits of Tibetan cashmere goats[J].Journal of Domestic Animal Ecology, 2014, 35(12):16-20.(in Chinese)
[37] 索朗达, 巴贵, 次仁德吉, 等.西藏绒山羊KAP7.1和KAP8.2基因多态性及其对部分生产性状的影响[J].畜牧与兽医, 2020, 52(2):13-17. SUO L D, BA G, CIREN D J, et al.Polymorphism of the KAP7.1 and KAP8.2 gene and their relationships with production traits of Tibetan cashmere goats[J].Animal Husbandry & Veterinary Medicine, 2020, 52(2):13-17.(in Chinese)
[38] 张亚妮, 张恩平, 吴迪, 等.KAP基因的多态性与辽宁绒山羊经济性状的关系研究[J].中国农业科学, 2007, 40(9):2062-2067. ZHANG Y N, ZHANG E P, WU D, et al.Studies of the relationship between polymorphism of KAP gene and economic traits for Liaoning cashmere goats[J].Scientia Agricultura Sinica, 2007, 40(9):2062-2067.(in Chinese)
[39] JIN M, CAO Q, WANG R L, et al.Molecular characterization and expression pattern of a novel keratin-associated protein 11.1 gene in the Liaoning cashmere goat (Capra hircus)[J].Asian-Australasian Journal of Animal Sciences, 2017, 30(3):328-337.
[40] JIN M, WANG L, LI S, et al.Characterization and expression analysis of KAP7.1, KAP8.2 gene in Liaoning new-breeding cashmere goat hair follicle[J].Molecular Biology Reports, 2011, 38(5):3023-3028.
[41] 杨雪梅, 温凌云, 何军敏.KAP16基因在苏博美利奴羊毛囊发育不同时期及不同组织中的表达分析[J].家畜生态学报, 2019, 40(10):28-32. YANG X M, WEN L Y, HE J M.Analysis of KAP16 gene expression in different stages of hair follicles development and different tissues in Subo Merino sheep[J].Journal of Domestic Animal Ecology, 2019, 40(10):28-32.(in Chinese)

内蒙古绒山羊KAP9.2基因遗传多态性及其与生长性状的关联分析

The Genetic Polymorphisms of KAP9.2 Gene and Its Correlation Analysis with Growth Traits of Inner Mongolia Cashmere Goats

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