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Analysis of MYB75and TT8Interaction on Anthocyanin Synthesis in Brassica Rapa Turnip

Author :WangGuanJie
Tutor:LiYuHua
School :Northeast Forestry University
CLC :Q945
TYPE :Masters thesis
Download the PDF Full Text:http://www.topresearch.org/showinfo-178-678043-0.html
Year:2013
Abstract:
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 OEBrTT8were 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.