二穗短柄草WRKY基因的表达分析及与MAPK的互作分析-制药工程专业论文.docxVIP

烯和双氧水诱导，推测乙烯和双氧水信号路径可能参与了这 5 个 BdWRKY 基因的表 达调控。 （4）构建 BdWRKYs 基因酵母表达载体 pGBKT7-BdWRKYs，与 pGADT7-BdMAPKs 质粒共同转化到酵母菌中进行互作分析，发现 BdWRKY26 与 BdMAPK7-1 之间存在 相互作用。 本文研究结果为深入探讨二穗短柄草 BdWRKY 与 BdMAPK 在响应非生物逆境 胁迫中的作用具有参考意义。 关键词：二穗短柄草，WRKY 基因，MAPK 基因，表达分析，酵母双杂交 Abstract WRKY proteins are transcription factor in eukaryotic cells that can directly combine with the promoter of the related genes, which can regulate gene expression. WRKY transcription factors have been reported to participate in seed development, bio- and abiotic stress response, and play important roles in abscisic acid, methyl jasmonate and salicylic acid signaling pathways. MAPK cascade pathway is highly conserved in the evolution of the plant，it transduces external signal to the nucleus by stepwise amplification process, so that the cells could respond to changes in the external environment. MAPK cascade plays an important role in plant responses to various bio- and abiotic stresses. It has been reported that MAPKs can interact with WRKY by means of phosphorylation, thus regulate WRKY activity and mediate various bio- and abiotic stress in the plants. Brachypodium distachyon is a new type of model plant species and its genome’s sequecing has been completed, but there is still no report on WRKYs in response to abiotic stress and the interaction with MAPKs. In this study, cDNA of five WRKY genes from B. distachyon were cloned, their expression changes under stress condition were analyzed by RT-PCR, and the interactions between BdWRKYs and BdMAPKs were exzamined by yeast two-hybrid technology. The results are as follows: Based on BdWRKYs sequences in NCBI database, full-length cDNAs sequences WRKY of five WRKYs were successfully clond using sequence-specific primers, and designed BdWRKY3、BdWRKY4、BdWRKY4-1、BdWRKY20 and BdWRKY26. Biological informatic analyses of the five BdWRKY gene sequences suggest that those BdWRKYs belong to the WRKY family members. RT-PCR analysis results of five BdWRKY gene expression under ab