Cloning And Analysis Of Dna And Promoter In Different Key Genes Involved In Flower Color Biosynthetic Pathway From Muscari

Muscari belonging to hyacinth division are bulb flowers in the spring, famous for its bright blue, and has very important aesthetics, ecological and scientific value. It is an ideal model for the study of the mechanism of color in Monocotyledons. This study bases on blue Muscari ‘Armeniacum’ and White Muscari ‘White beauty’ for material and bases on the transcriptome sequencing results and papers has been published by our team, cloning DNA sequences and promoter sequences of these seven proteins,synthase(CHS), chalcone isomerase chalcone(CHI), flavanone once enzyme(F3H), flavonoid 3’5’-hydroxylase(F3’5’H), dihydrogen flavonols reductase(DFR),anthocyanins synthetase(ANS/LDOX) and flavonol synthase(FLS), in blue and white varieties to analyze each gene of between blue and white varieties,to determine ultimately the significantly different genes. Following as main results:1. Based on blue Muscari ‘Armeniacum’ and white Muscari ‘White beauty’ for material to clone these key genes in anthocyanin pathway as CHS, CHI, F3 H, F3’5’ H, DFR, FLS, ANS and their promoter sequences, I have obtained successfully DNA sequences having completed conserved domain and promoter sequences of 7 genes above.2. I have compared these DNA sequences and their protein cloned in the blue and white varieties.I found that the similarity of CHS sequences is minimal(88.41%)of all and their protein similarity is 34%. The similarities of Other DNA sequences and proteins are all above 90% and their difference between blue and white varieties are minor. The differences of promoters between blue and white varieties more obvious than DNA. The similarity of DFR promoter between blue and white varieties is 90.52% and CHI promoter is 84.55%.Other promoters are below 75%. F3 H and FLS promoter are as low as 52.9% and 52.9% respectively.3. I have analyzed these promoters in the PlantCARE database and estimated the function and quantity of components on promoters.Comparing to the number of elements on each promoter from blue and white varieties, I found ARR1ATG8: G11 and GATABOX are different significantly in the varieties of blue and white,followed by POLLEN1LELAT52, IBOXCORE,SORLIP2 AT,other elements had smaller variations. ARR1ATG8: G11 elements from DFR, FLS,F3 H and F3’5’H promoters and GATABOX elements from CHS, FLS, ANS and CHI promoters are different significantly.4. I have analyzed bHLH binding sites and MYB binding sites of these promoters, I found bHLH binding sites and MYB binding sites of CHS, F3 H and FLS promoters in blue and white varieties are large differences, bHLH binding sites on CHS promoters in are two more than ‘White beauty’, bHLH binding sites and MYB binding sites of F3 H promoter in White beauty are one more than Armenia respectively,MYB binding sites of FLS promoter in the White beauty are two more than Armenia.