Cloning And Expressin Analysis Of Anthocyanin Biosynthesis Related Genes In Cerasus Humilis

Cerasus humilis is a Special fruit resources in our country, classified as Rosaceae Ceraras. Cerasus humilis fruit has bright color, unique flavor, rich nutrition, especially fruits rich in anthocyanins, flavonoids and carotenoids and other pigments, research and development and utilization has a very high value. In this paper, chlorophyll, carotenoid, anthocyanin and flavonoid content in’Nongda No.4’and ’Nongda No.5’were determined. The cDNA sequences of CHS、CHI、F3H、DFR、ANS、UFGT genes in Cerasus humilis were cloned by homologous cloning, and then by using fluorescence quantitative techniques to study the expression of the Cerasus humilis during development of the gene CHS、CHI、 F3H、DFR、ANS、UFGT. The main results are as follows:(1)The main anthocyanins of Cerasus humilis is Cyanidin-3-O-glucoside and Pelargonidin-3-O-glucoside. In the early stage of ’Nongda No.4’ and ’Nongda No.5’ fruit development,the content of chlorophyll is high and other pigments content is low,so the mainly pigments is chlorophyll.The anthocyanin in fruit development of ’Nongda No.4’ is gradually increased to the mature fruit content reached the highest point.carotenoid content decreased first and then increased, the content of chlorophyll decreased gradually to the lowest,so the mainly pigments is anthocyanin.The chlorophyll in fruit development of ’Nongda No.5’ is gradually decreased,carotenoid content decreased first and then increased to reach the highest,anthocyanin content has been at a low level,so the mainly pigments is carotenoid.(2) The full-length cDNA sequences of ChCHS、ChCHI、ChF3H、ChDFR、ChANS and ChUFGT genes in Cerasus humilis were cloned. The length of that genes were 1176 bp,651 bp,1086 bp,1041 bp, 1074 bp and 1428 bp. Using bioinformatics analysis, speculated that genes encode 391、216、361、346、 357and475 amino acids, with molecular mass of 42.77、23.21、40.41、38.84、40.40 and 51.59 kDa. With multiple comparisons to the amino acid sequence of others species related genes,the similarity of each gene and related genes of Prunus avium more than 95%.(3) The results of real-time PCR showed that the expression of ChCHS, ChCHI、ChDFR、ChANS and ChUFGT in peel of nongda 4 were less in early fruit development, up-regulated expression in veraisom and then expression declined. The expression of ChF3H in peel of nongda 4 was rised in the fruit development. The expression of ChCHS、ChF3H、ChDFR、ChANS and ChUFGT in pulp of nongda 4 were less in Early fruit development, up-regulated expression in veraisom and then expression declined. The expression of ChCHI in pulp of nongda 4 was rised in the fruit development. The expression of ChCHS、 ChCHI、ChF3H、ChDFR、ChANS and ChUFGT in peel and pulp of nongda 5 was expressed in low abundance in the fruit development.