Functional Anlaysis Of The Key Transcription Factors Invovled In The Blue Flower Formation In Muscari Botryoides

Grape hyacinths(Muscari botryoides) is a liliaceous plants of the genus blue pot of flowers and a kind of imported from European perennial herbaceous flower flower. In the plant transcription factor, MYB transcription factor is one of the largest and most widely research family number. MYB transcription factors are widely involved in regulating cell growth and differentiation, the external environment and plant hormone response and regulation of plant flavonoids metabolism. In recent years, the pear, gentiana, lily has related gene cloning and identified in plants, such as the MYB transcription factors, and about cloning analysis of transcription factors in grape hyacinths rarely reported. The study clones a new MYB transcription factor gene from the grape hyacinths to bioinformatics and expression pattern analysis. In addition, the blue pigment content in grape hyacinths was determined, and the transcription factor gene expression has carried on the correlation analysis, the results are as follows:1.According to the grape hyacinths transcriptome sequencing result, choose high expression of candidate genes. Design specific primers and get the transcription factor gene through RACE-PCR, with Ma MYB2 named. Sequence analysis, obtained the open reading frame of 600 bp, the gene encoding 199 amino acids, speculated that its coding protein molecular weight of about 22.34 k D, and petunias homology, the highest is 82%; With lily, columbines, violet in the summer, apple, tomato, strawberry, dahlia, hula and other 18 kinds of plant MYB transcription factors of homology in 43% to 82%. The grape hyacinths Ma MYB2 protein sequence analysis found that is typical in the conservative HTH structure can be combined with a DNA region, indicating it is likely to have transcription regulation effect.2.The real time quantitative PCR results show that the grape hyacinths‘M.armeniacum’are detected in different tissue parts Ma MYB2 gene expression. And Ma MYB2 in flower development of S5 expression quantity is higher than the others. The result is in line with the determination of the content of anthocyanins.3.Build yeast expression vector p GBKT7-Ma MYB2 into yeast strains Y2 HGOLD, X-α-gal in containing three short of medium(SD /- Trp- His- Ade + X- alpha gal), the results showed that the Ma MYB2 in yeast can grow and show blue with transcriptional activation activity. Make PBI221-MYB-GFP recombinant plasmid into onion skin cell. The results showed that the protein localization in the nucleus. Yeast activation and verification test show that the Ma MYB2 has the basic characteristics of transcription factors.4.Different tissue parts was carried out on the blue varieties‘M.armeniacum’flower pigment content determination, the results show that grape hyacinths almost undetectable in root, stem and leaf of anthocyanin accumulation. S1 contains only a small amount of cornflower pigment. S2-S5 period have a lot of delphinidin anthocyanin detected. And gene transcription factor correlation analysis found a positive correlation. Correlation analysis was carried out on the genes and gene transcription factors, finding Ma MYB2 and ANS, DFR, F3 ’5’ H correlation, presumably the transcription factors likely to manage through with these genes anthocyanin accumulation.