调控甘草黄酮生物合成的MYB转录因子的筛选与功能研究

项目名称： 调控甘草黄酮生物合成的MYB转录因子的筛选与功能研究

项目编号： No.31460064

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 李雅丽

作者单位： 内蒙古科技大学

项目金额： 50万元

中文摘要： 转录因子可激活特定次生代谢产物系列合成酶的协同表达，促进该类产物的大量合成，因而利用转录因子构建高产转基因细胞系是解决目前植物细胞培养产量低的有效途径。黄酮类化合物作为甘草中的主要活性成分，其生物合成途径已非常明晰，但调控黄酮类合成的分子及其调控机制尚不清楚，本项目以鄂尔多斯高原甘草细胞为研究对象，在对经茉莉酸甲酯（MeJA）诱导前后的甘草悬浮细胞进行转录组高通量测序，通过比较发现12个可能参与甘草黄酮类合成相关的MYB转录因子的基础上，拟以甘草黄酮合成途径中的关键基因chi启动子为诱饵进行酵母单杂交实验，确定参与调控甘草黄酮合成的MYB转录因子，将其转化到甘草细胞中进行过表达及特异性干涉，并利用DNaseI足迹法和EMSA确定其与chi基因启动子的结合位点，确定参与调控甘草黄酮合成的MYB转录因子及其调控效应，为推动甘草细胞大规模培养的工业化进程奠定基础。

中文关键词： 转录因子；甘草黄酮；甘草；生物合成

英文摘要： Transcription factors can activate a series of synthetase producing specific secondary metabolites to coexpress and promote the product synthesis, so it is an effective ways to solve the problem of low yield in plant cell culture by using transcription factor to construct a transgenic cell lines of high yield. As the main active ingredient in Glycyrrhiza, flavonoids biosynthetic pathway has been very clear,but the regulation molecules and regulation mechanism of flavonoids synthesis is unclear.In this study, Glycyrrhiza cell from Erdos plateau as the object,High-Throughput Transcriptome Sequencing for MeJA treatment and mock-treated licorice suspension cells had found 12 transcription factors related to Glycyrrhiza flavonoids synthesis.It is proposed Chi gene (the key genes in the biosynthesis of flavonoids) promoter as bait for yeast one hybrid experiment to determine the MYB transcription factors involved in the regulation of flavonoid synthesis. Then the MYB transcription factors were transformed into cells overexpressing and specificity interference, and identified the binding sites with chi gene promoter using DNaseI footprinting and EMSA. After that, regulation effect of these MYB transcription factors to flavonoid synthesis and chi gene expression would be explored.By executing this project,it is expected to solve the problem of low yield and lay the foundation for promoting the industrialization process of large-scale cultivation of licorice cells.

英文关键词： Transcription factors;Glycyrrhiza flavonoids;Glycyrrhiza;Biosynthesis