|
|
Correlation Analysis of ALX4 Gene Polymorphism and Cashmere Traits in Cashmere Goats (Capra hircus) |
ZHANG Zhi-Long1,2, WU Tie-Cheng2, LI Yu-Rong2, MA Yue-Jun2, ZHAO Xia2, ZHAO Sheng-Guo1, LIU Bin2,*, CAI Yuan1,* |
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 Animal Husbandry Institute of Inner Mongolia Autonomous Region Academy of Agricultural and Husbandry Sciences, Hohhot 010031, China |
|
|
Abstract During the development of cashmere goat (Capra hircus), different gene expression regulation plays an important role in hair follicle metabolism. In this study, ALX4 (aristaless-like homeobox 4) was selected as the candidate gene to verify the correlation between ALX4 gene and cashmere goat traits, and to provide a basis for molecular marker-assisted selection of cashmere goats. The blood DNA of 20 cashmere goats with maximum and minimum was first extracted, and the 3'-UTR, 5'-UTR, exon, and promoter sequences of the ALX4 gene were amplified and sequenced by PCR amplification technology. According to the results of Blast comparison, SNP loci with mutations in ALX4 gene were counted, and 315 cashmere goat individuals were detected using KASP (kompetitive allele-specific PCR)genotyping technology. The correlation analysis between SNPs loci and villi traits was performed using a general linear model. The results showed that SNP1 (T-1700C) exists in the promoter region of ALX4 gene, SNP2 (T-14621C), SNP3 (T-15433C), SNP4 (G-16377C), SNP5 (A-17780G) and SNP6 (A-18044G) in the 3'-UTR. Among them, the frequency of CG genotype of SNP4 locus of ALX4 gene is the highest (0.4459), which is the dominant genotype, and the G allele is the dominant allele; the frequency of GG genotype of SNP6 locus is the highest (0.5641), which is the dominant genotype, and the G allele is the dominant allele. All SNP loci were in Hardy-Weinberg equilibrium in cashmere goat population (P>0.05), and the homozygosity was higher and the heterozygosity was lower in the population. The effective allele of SNP4 (1.8947) was the highest among all SNPs, and that of SNP1 was the highest. The number of effective alleles (1.2534) was the lowest than other SNPs; except for SNP1 in low polymorphism (PIC<0.25), all other SNPs loci were in moderately high (0.5>PIC>0.25). Correlation analysis showed that the SNP4 locus of ALX4 gene was significantly associated with wool length traits (P<0.05), the wool length of individuals with GG genotype was significantly higher than those of CC and CG individuals (P<0.05); the SNP6 was significantly associated with cashmere fineness traits (P<0.05) the cashmere fineness of AA individuals was significantly higher than that of GG and AG individuals (P<0.05); A total of 12 haplotypes were constructed using PHASE software, with 4 haplotypes having a frequency>5%, which were: H1 (TCCAA, 20.384%), H4 (TCCGG, 15.029%), H7 (TCGGG, 38.498%), and H12 (CTGGG, 22.248%). In the haplotype combination of 5 SNPs of the ALX4 gene, there were 9 haplotype combinations with a frequency>5%. Correlation analysis of these 9 haplotype combinations with cashmere production traits indicated that different haplotype combinations of 5 SNPs in the 3'-UTR region of ALX4 gene were significantly associated with wool length correlation (P<0.05), the wool length of H1H1 individuals was significantly higher than that of HIH7, H1H12, H4H7 and H7H7 (P<0.05). The above results indicate that the ALX4 gene can be used as a key gene for molecular marker-assisted breeding in cashmere goats. In the breeding work of cashmere goats, individuals with AG genotypes at the SNP6 locus can be selected for ultrafine cashmere goat breeding.
|
Received: 17 December 2019
|
|
Corresponding Authors:
*liubin0613@126.com;caiyuan@gsau.edu.cn
|
|
|
|
[1] 惠太宇,郑圆媛,岳畅,等.2019.羊绒细度候选基因靶标miRNAs筛选和鉴定[J].沈阳农业大学学报,50(01):19-27.
(Hui T Y,Zheng Y Y,Yue C,et al.2019.Screening and identification of cashmere fineness candidate gene targets miRNAs[J].Journal of Shenyang Agricultural University,50(01):19-27.)
[2] 丽春,张文广,杨文柱,等.2012.褪黑激素对PDGFA基因在绒山羊皮肤毛囊中表达模式的影响[J].中国农业科学,45(10):2031-2039.
(Li C,Zhang W G,Yang Y Z,et al.2012.Effects of melatonin on the expression pattern of PDGFA gene in the skin and hair follicles of cashmere goats[J].Chinese Agricultural Sciences,45(10):2031-2039.)
[3] 李学武.2019.内蒙古绒山羊毛被类型遗传规律及其对重要经济性状间接选择研究[D].博士学位论文,内蒙古农业大学,导师:李金泉.pp.62-65.
(Li X W.2019.Study on the genetic law of Inner Mongolia cashmere wool quilt type and its indirect selection of important economic traits.[D] Thesis for Ph.D.,Inner Mongolia Agricultural University,Supervisor:Li J Q.pp.62-65.)
[4] 王利生,翁立强,程波.等.2007.性激素影响毛囊生长的机制进展[J].国际皮肤性病学杂志,33(05):311-313.
(Wang L S,Wen L Q,Cheng B.2007.Mechanism of sex hormones affecting hair follicle growth[J].International Journal of Dermatology,33(05):311-313.)
[5] 魏云霞,肖玉萍,杨保平,等.2012.河西绒山羊绒毛生长的季节性变化规律及其与生长激素关系的研究[J].中国草食动物科学,32(06):10-13.
(Wang Y X,Xiao Y P,Yang B P,et al.2012.A study on the seasonal variation of the growth of Hexi cshmere goat and its relationship with growth hormone[J].Chinese Herbivore Science,32(06):10-13.)
[6] 尹俊.2004.内蒙古绒山羊毛囊发育、生长周期及相关基因的研究[D].博士学位论文,内蒙古大学,导师:扈廷茂.pp.3-7.
(Yin J.2004.Study on development,growth cycle and related genes of wool fleece in Inner Mongolia[D].Thesis for Ph.D.,Mongolian University,Supervisor:Hu T M.pp.3-7.)
[7] 袁海旭,王明月,赵越,等.2018.鸡主要组织相容性复合体B区域多态性分析[J].中国家禽,40(20):8-13.
(Yuan H X,Wang M Y,Zhao Y,et al.2018.Polymorphism analysis of chicken major histocompatibility complex B region[J].Chinese Poultry,40(20):8-13.)
[8] Borasgranic K,Grosschedl R,Hamel P A.2004.Genetic interaction between Lef1 and Alx4 is required for early embryonic development[J].International Journal of Developmental Biology,50(7):601-610.
[9] Ebert M P A,Model F,Mooney S,et al.Aristaless-like2006.homeobox-4 gene methylation is a potential marker for colorectal adenocarcinomas[J].Gastroenterology,131(5):1418-1430.
[10] Jinxing H,Xiaopeng A,Yuxuan S,et al.2015.Two mutations in the caprine MTHFR 3'UTR regulated by microRNAs are associated with milk production traits[J].PLOS ONE,10(7):e0133015.
[11] Hamel.2006.Loss of Alx4,a stromally-restricted homeodomain protein,impairs mammary epithelial morphogenesis[J].Developmental Biology,297(1):0-294.
[12] Kulessa H,Turk G,Hogan B L.2000.Inhibition of Bmp signaling affects growth and differentiation in the anagen hair follicle[J].The EMBO Journal,19(24):6664-6674.
[13] Lee Y R,Yamazaki M,Mitsui S,et al.2001.Hepatocyte growth factor (HGF) activator expressed in hair follicles is involved in in vitro HGF-dependent hair follicle elongation[J].Journal of Dermatological Science,25(2):156-163.
[14] Millar S E.2002.Molecular mechanisms regulating hair follicle development[J].Journal of Investigative Dermatology,118(2):216-225.
[15] Nishimura Emi K,2011.Melanocyte stem cells:A melanocyte reservoir in hair follicles for hair and skin pigmentation[J].Pigment cell & Melanoma Rresearch,24(3):401-410.
[16] Richardson G D,Bazzi H,Fantauzzo K A,et al.2009.KGF and EGF signalling block hair follicle induction and promote interfollicular epidermal fate in developing mouse skin[J].Development,136(13):2153-2164.
[17] Ritva R,David P C,Rice B,et al.2003.Progression of calvarial bone development requires Foxc1 regulation of Msx2 and Alx4[J].Developmental Biology,262(1):75-87.
[18] Roh J,Pi W.2013.Regulation of follicle growth and apoptosis in cultured mouse ovaries by NGF and oxygen[J].Gynecological Endocrinology,29(6):574-579.
[19] Shi Y,Sun X,He X.2017.Overexpression of aristaless-like homeobox-4 inhibits proliferation,invasion,and EMT in hepatocellular carcinoma cells[J].Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics,25(1):11-18.
[20] Shu N,Jia-Qi C,Shu-Yuan G,et al.2018.Expression profile of fibroblast growth factor 7 sub-family in the first hair follicle cycle of mice skin[J].Chinese Journal of Animal and Veterinary Sciences,49(3):515-524.
[21] Stark A,Brennecke J,Bushati N,et al.2005.Animal microRNAs confer robustness to gene expression and have a significant impact on 3'UTR evolution[J].Cell,123(6):1122-1146.
[22] Wuyts W.2000.The ALX4 homeobox gene is mutated in patients with ossification defects of the skull (foramina parietalia permagna,OMIM 168500)[J].Journal of Medical Genetics,37(12):916-920.
[23] Yang,Han J T,Fei L,et al.2017.ALX4,an epigenetically down regulated tumor suppressor,inhibits breast cancer progression by interfering Wnt/β-catenin pathway[J].Journal of Experimental & Clinical Cancer Research,36(1):162-170.
[24] Yano K,Brown L F,Detmar M.2001.Control of hair growth and follicle size by VEGF-mediated angiogenesis[J].Journal of Clinical Investigation,107(4):409-417. |
|
|
|