Analysis Of MYB75and TT8Interaction On Anthocyanin Synthesis In Brassica Rapa Turnip

Transcription factors play important roles in the regulation of plant growth and development. The transcription factors can repress or activate the expression of genes by interacting with other proteins and binding DNA through the functional domains. MYB and bHLH transcription factors are important protein families in plant genes, anthocyanin synthesis and adapt to biological or abiotic stress. In this study, Tsuda, which is a light-dependent anthocyanin biosynthesis plant, was used in our experiment. The BrMYB75and BrTT8transcription factors have been cloned in the lab, using subcellular localization to determine its role in intracellular location. To determine the regulation function of BrMYB75and BrTT8proteins in anthocyanin biosynthesis, the yeast two hybrid, genetic transformation and other experiments were used to research. The results provide the basis for the futher study of MYB and bHLH transcription factors. The main results were obtained as follows.1:MYB75and TT8transcription factors associated with plant anthocyanin synthesis,and the tissue specificity and light induced specific expression of MYB75is consistent with TT8.2:The green fluorescent protein expression vectors GFP-BrMYB75and GFP-BrTT8were constructed successfully.Transformed to the epidermal cells, the result showed that both BrMYB75and BrTT8were loealized in cell nucleus.3:The expression vectors YEHBD-MYB75and Y2HAD-TT8were constructed successfully, transformed the competence of yeast. The yeast plaque appear blue which transformed both YEHBD-MYB75and Y2HAD-TT8. The test verifies the interaction between BrMYB75and BrTT8transcription factors.4:The overexpression vectors OE-BrMYB75and OE-BrTT8were constructed successfully, mix with BrCHS agrobacterium strains contain the luciferase agrobacterium strains transient transfection of tobacco leaf.Agrobacterium strains of BrCHS, BrMYB75and BrTT8, transient expression analysis of co-transformed tobacoo leaves illuminated that CHS was regulated by BrMYB75and BrTT8.5:With Gateway clonging system, expression vector, pCAMBIA1301-PMI-BrMYB75and pCAMBIA1301-PMI-BrTT8were constructed and transformed into tomato and tobacco. The combination of PMI seleetion medium could be used to obtain the adventitious bud. Cut the adventitious bud for rooting cultivation, and selected the positive plants. Trans genie tomato and tombacoo plants have been obtained and some of them were PCR positive.In plant photoinduced anthocyanin synthesis route, through the study of this paper have made a preliminary proof that the regulation of MYB75and TT8transcription factors on CHS gene anthocyanin structure synthesis, but the function of MYB75and TT8in plants on anthocyanin synthesis pathway in plant also requires further study.