Screening And Functional Study Of Fox Fur Color Differentially Expressed Genes

Foxes are divided into genus Vulpes vulpes foxes and Alopex lagopus foxes,which have diverse coat colors.Screening for coat color regulation genes and studying gene expression are conducive to revealing the mechanism of fox coat color formation and mutation,and promoting fur animal breeding and molecular breeding progress.RNA-seq was used to detect the sequences of five fox species including arctic blue fox,arctic white fox,red fox,silver fox and silver cross fox.After data processing and Long sequence clustering,378741 All-unigenes were obtained,of which the total length is 288149813 nt,the average length is 761 nt,N50 is 1530 nt.Through function and pathway analysis,34 genes including MC1 R,Agouti,MITF and so on related to fox coat color were marked in the KEGG melanogenesis pathway.Then,the real-time quantitative PCR technology was used to verify the three differentially expressed genes(MITF,MC1 R,and Agouti)in melanogenesis pathway,and the two genes(CBD103 and PMEL)with differential expression multiple largely in the transcriptome results that have important effects on coat color in other species.High-throughput sequencing results showed that Genes such as MITF and Agouti are down-regulated genes;MC1R,PMEL and so on are more expressed in red fox than other fox species.The real-time quantitative PCR results showed that 5 genes were expressed in different tissues(skin,heart,liver,spleen,lung,kidney and muscle)of arctic foxes,and the highly expressed in the skin.Comparing the mRNA expression levels of different foxes including artic blue fox,artic white fox,red fox and silver black fox,the results showed:(1)MITF is the highest expressed in the skin of Arctic white fox,followed by Arctic blue fox,red fox and silver black fox.(2)MC1R gene expressed in red fox skin mRNA is significantly higher than other foxes(P <0.05),followed by Arctic blue fox,and Arctic white fox and silver black fox.(3)The expression level of Agouti is highest in Artic white fox,and followed by Arctic blue fox,red fox and silver black fox.(4)CBD103 is the highest expressed in the skin of red fox,followed by Arctic wtite fox and Artic blue fox,while silver black fox is lowest.(5)The expression of PMEL in red fox is signicificantly higher than others(P <0.05),followed by Artic blue fox,Artic white fox and silver black fox,the difference of which is not signicifant.The mRNA expression levels of five genes in the arctic blue fox and arctic white fox skin tissues were the highest,and the mRNA expression results in the skin tissues of different coat foxes were basically consistent with the transcriptome sequencing results.And we get the conclusion: the expression of MITF and Agouti gene gradually decreased as the coat color deepened.MC1 R,CBD103 and PMEL genes were highly expressed in red foxes,from which we speculate that promoted the formation of brown phenotype..