| The spotted horse is a unique color type of Kazakh Aletai type cultivated in the history of Kazakh.This study uses 60 spotted horses and 12 Khazak horses,the KIT gene exon 21 variant polymorphism analysis and bioinformatics analysis,using real-time fluorescence quantitative PCR of 5 kinds of spotted horses skin tissue KIT,MITF,EDNRB in mRNA expression analysis.The results of the test are as follows:(1)The expression of KIT gene is different in the skin tissue of spotted horses.The expression level of KIT gene according to the sequence showing white greys > white spots1 >white spots2 >brown spots >chestnut spots,the expression of KIT gene was the highest in the skin tissues of white grey horses.In the same skin tissue,white spot1 > brown spots,white spot 2 > chestnut spots.There was significant difference between the expression in white grey hair and that in the skin of chestnut spots and brown spots and the lowest expression was found in the skin of chestnut spots.(2)The expression of MITF gene is different in the skin tissue of spotted horses.The expression level of MITF gene according to the sequence showing brown spots >chestnut spots > white spots1 > white spots2 >white greys,the expression of MITFgene was the highest in the skin tissues of brown spots.In the same skin tissue,brown spots >white spot1,chestnut spots > white spot 2.There was significant difference between the expression in white greys hair and that in the skin of chestnut spots,brown spots,white spot1,white spot2 and the lowest expression was found in the skin of white greys hair.(3)The expression of EDNRB gene is different in the skin tissue of spotted horses.The expression level of EDNRB gene according to the sequence showing white spots2 >white spots1 >white greys >chestnut spots >brown spots,the expression of EDNRB gene was the highest in the skin tissues of white spots2.There was significant difference between the expression in white spots2 and that in the skin of chestnut spots,brown spots,there wassignificant difference between the expression in white spots1 and that in the skin of chestnut spots,brown spots,in the same skin tissue,white spot1 > brown spots,white spot 2 >chestnut spots,the lowest expression was found in the skin of brown spots.(4)The three genes of KIT,MITF and EDNRB were expressed in different degrees in the skin tissue of spotted horse.The expression of EDNRB gene was the highest in every skin tissue.The EDNRB gene was significantly different from the KIT and MITF genes in white spot1,white spot 2,brown spots and chestnut spots(P < 0.05).There was no significant difference between the EDNRB gene and the KIT gene in the white greys,and there was a significant difference from the MITF gene(P< 0.01).The expression of EDNRB was highest in all skin tissues,especially in white spots and white greys.(5)The exons of the hairy spot KIT gene of Kazakh horse were sequenced.the first exon of KIT gene was found to be SNP site g.210 T > G trans version,which was transformed from amino acid encoded leucine(CGC)to arginine(CUC).So there is a wrong mutation in the g.210 T > G site,by chi square test,it was shown that the effect of the mutation site on the color of white spots hair was significant(P< 0.01).Bioinformatics analysis showed that Pi =0.000505,indicating that the KIT gene of spotted horse is a relatively conservative gene,Tajima'D =-1.695842(P >0.05).Phylogenetic tree analysis of all samples,Fuyun County,Qinghe County,the spotted horse genetic distance,fully consistent with the geographical location,genetic distance between Aletai city and Fuhai County far.There is no special genetic distance between the spotted horses in each area,and all of them belong to a group of Kazakh horses.In conclusion,The mutation of the KIT gene may be associated Of spotted horses.The MITF gene is brown spots,and KIT and EDNRB are the candidates for white spots and white greys,In particular,EDNRB is the candidate gene that has the greatest impact on white spots. |
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