苦荞R2R3-MYB SG7类转录因子基因功能分析

R2R3-MYB SG7类转录因子家族包括MYB11、MYB111和MYB12三个成员，对拟南芥黄酮醇类物质合成起着重要的调控作用。已有研究表明苦荞含有高活性黄酮醇类物质-芦丁。我们前期研究工作证实苦荞品种间芦丁含量存在差异，且外源生理生化因子影响其芦丁的合成及结构基因表达变化，为进一步揭示苦荞黄酮醇类物质合成的转录调控机制，我们拟从苦荞中克隆调控黄酮醇类物质合成的R2R3-MYB SG7类转录因子，进行功能分析：（1）keypoint技术筛选苦荞myb11、myb111和myb12突变体，比较分析黄酮醇含量变化部位及结构基因表达差异，明确该类转录因子调控的靶基因及调控部位；（2）EMSA技术验证转录因子与靶基因启动子的相互作用；酵母单杂技术验证转录因子的转录激活特征。上述结果将揭示苦荞R2R3-MYB SG7类转录因子参与黄酮醇类物质的转录调控机制，为苦荞分子育种提供科学依据。

R2R3-MYB SG7 transcription factors play important roles in the regulation of the biosynthesis of flavonls in Arabidopsis thaliana.There are three members including MYB11,MYB111 and MYB12 in R2R3-MYB SG7 family. F.tartarcium contains highly active flavonls such as rutin.In our perious work, the content of flavonls in different cultivars of F.tartarcium varied to a great extent, and treatments with exogenous physiological and biochemical factors such as UV-C, Salicylic Acid and Phenylalanine afftected the the rutin content and the expression level of key genes in the flavonl biosynthesis pathway. In order to understand the regulation mechanism of flavol biosynthesis in F.tartarcium, we propose here to clone and study the functions of the genes of R2R3-MYB SG7 transcription factors:(1) Screening F.tartarcium EMS mutagenesis library for the mutants of R2R3 MYB-SG7 transcription factors (myb11,myb11,myb12) by keypoint technonlogies;Comparative analysis the content of flavonls and the expression level of structual genes,then the target gene and regulatory aera will be idetified.(2) By electrophoretic mobility shift assay（EMSA）could be identified the interaction between promotor of target sequence in R2R3 MYB SG7 transcription factors and regulatory aera.Yeast one-hybrid technology to understand the regulation mechanism of R2R3 MYB SG7 transcription factors in flavonols biosynthesis.The results dicovered the regulatory mechnism of R2R3-MYB SG7 transcription factors,thus providing the scientific basis for the molecular breeding of F.tartarcium.