Cloning And Sequence Analysis For Anthocyanin Transcriptional Activator Genes Of Potato

Anthocyanin has some important physiological functions, such as antioxidation, antimutation, antiproliferation etc. Purple potato is intituled healthy food for its abundant anthocyanin. Pigmentation patterns and the intensity of anthocyanin in various plant tissues are coordinately controlled by the expression of anthocyanin transcriptional activator genes in plants. Cloning and identification of anthocyanin transcriptional activator genes of potato (MYC and MYB like genes) are very useful for elucidation the mechanisms that control the patial and temporal pigmentation in potato and for controlling the pigmentation in potato by using the anthocyanin transcriptional activator genes as switches.In this study, the conserved 5–end fragment (Stmyc1) of myc like anthocyanin transcriptional activator gene was amplified by using RT–PCR from the RNA of purple potato coat. Then the cDNA 3-end sequence (Stmyc2) was amplified by using 3'-Race technique from the RNA of purple potato coat. The myc like anthocyanin transcriptional activator gene (Stmyc) of potato was got by connecting Stmyc1 fragment and Stmyc2 fragment. The length of Stmyc gene is 1106bp, containing an intact ORF and coding 326aa.At the same time, the conserved fragment (Stmyb1) of myb like anthocyanin transcriptional activator gene was amplified by using RT–PCR from the RNA of purple potato coat. They belong to the R2R3 myb conserved fragment (named Stmyb280 and Stmyb211) of myb like anthocyanin transcriptional activator gene. Conclusion:1) We isolated the myc like anthocyanin transcriptional activator cDNA (Stmyc) from purple potato coat. The length of it is 1106bp, Which contained an intact ORF. The ORF located between positions 10 and 990 and coding 326aa. After sequence analysis, The deduced protein of Stmyc gene is similar to that of Perilla frutescens, the similarity is 94%. Stmyc gene encodes a bHLH protein domain that is more important for MYC-like proteins. So we speculate that Stmyc gene is correlated with the anthocyanin transcriptional activator myc like gene.2) We isolated the conserved fragments of R2R3 myb like anthocyanin transcriptional activator gene from purple potato coat. The deduced protein of Stmyb211 fragment shares the most similarity with X95296 from L.esculentum, DQ277683 from Vitis vinifera, BT006302 and AY519615 from Arabidopsis thaliana.