相比于生物合成基因，转录因子对代谢途径的调控能力更显著，在基因工程改良方面更具前景。调控紫杉醇生物合成的转录因子的研究甚少。近期，申请人所在课题组对中国红豆杉细胞进行转录组测序，发现了两个不同类型WRKY类转录因子TcWRKY2(Ｉ型）、TcWRKY6（II型）在MeJA处理前后表达差异极显著，序列分析表明这两个基因是新成员。本项目拟对这两个WRKY类转录因子的功能进行深入研究，最终全面明晰WRKY类转录因子对紫杉醇生物合成的调节作用及调控机制。主要包括：两个WRKY类转录因子基因受诱导表达特点和亚细胞定位分析；利用超量表达、RNA干涉等技术深入解析两个基因功能及调控的生物学途径；利用染色质免疫共沉淀-深度测序（ChIP-seq）及凝胶阻滞（EMSA）技术研究两个WRKY类转录因子调控的靶基因。研究结果不仅可以丰富紫杉醇生物合成的转录调控理论，还将为稳定高产红豆杉细胞系的创建提供依据。

The regulation of transcription factors on plant secondary metabolism has become hot spot both at home and abroad, the regulation of transcription factor on metabolic pathway is more comprehensive and more notable compared with biosynthesis genes in the genetic engineering improvement. The research on the transcriptional factor regulated taxol biosynthesis is scarce. Recently, the applicant found two WRKY　transcriptional factor (TcWRKY2 and TcWRKY6) which can be induced by MeJA significantly. The sequence analysis showed these two genes are new member of type I and type II WRKY transcriptional factor. In this project, the founctions of these two genes were deeply researched, and the transcriptional regulation mechanism of WRKY transcriptional factor on taxol biosynthesis would be clarified. The main content including: induced expression characteristics by MeJA,SA,F5 fungi inducer and subcellular localization of these two transcriptional factor; clarify the founctions and regultated biology pathway of these two genes by over expression and RNA interference technology; clarify the target gene of TcWRKY2 and TcWRKY6 by ChIP-seq and EMSA technology. The research results can not only enrich the transcriptional regulation theory of taxol biosynthesis, also can provide the evidence for the genetic engineering improvement of Taxus cell.