柽柳WRKY转录因子应答外源ABA的分子调控机制

项目名称： 柽柳WRKY转录因子应答外源ABA的分子调控机制

项目编号： No.31200498

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 林学

项目作者： 郑磊

作者单位： 东北林业大学

项目金额： 22万元

中文摘要： WRKY转录因子是近年来发现的一类植物特有的新型转录因子。WRKY基因受多种环境因子诱导表达，与植物的生物和非生物胁迫应答密切相关。本研究从抗逆植物柽柳中克隆WRKY基因，将其转入拟南芥中，筛选对ABA抗性较强的株系进一步研究。利用cDNA微阵列技术筛选转WRKY基因拟南芥与野生型拟南芥的差异表达基因，从而确定可能受WRKY转录因子调控的下游靶基因。分析这些靶基因的启动子序列，通过酵母单杂交鉴定出可能与WRKY作用的顺式作用元件。将在胁迫响应过程中起重要作用的启动子区段构建到pHis2报告载体中，利用酵母单杂交技术研究在这段序列上起调控作用的基因。通过进一步验证，确定在抗逆过程中调控WRKY表达的上游基因。通过上述研究阐明WRKY转录因子应答ABA胁迫的分子调控机制。

中文关键词： 非生物胁迫；ABA；基因表达调控；刚毛柽柳；WRKY转录因子

英文摘要： WRKY transcription factor is a class of new plant-specific transcription factor that are found in the recent years. The expression of WRKY is found to be induced by various of environmental factors, and is closely related with the biotic and abiotic stress responses. In this project, a WRKY gene is cloned from Tamarix hispida which is extremely resistant to stresses. The WRKY gene is transferred to Arabidopsis. The transgenic line with the strongest resistance to ABA is screened for further study. The differentially expressed genes between WRKY transgenic Arabidopsis and wild-type Arabidopsis will be monitored by using cDNA microarray technology. Consequently, the potential downstream target genes that may be regulated by WRKY transcription factor can be confirmed. Some cis-acting elements in common in these target promoters are confirmed by sequence analysis using bioinformatic program. Cis-acting elements potentially bind to WRKY transcription factor are identified by yeast one-hybrid method. Promoter segments which play an important role in the stress response process is constructed into reporter vector pHis2, and the genes bind with this promoter segment are determined by using yeast one-hybrid system . Further validation is performed to determine the upstream regulatory genes of WRKY in the stress resistanc

英文关键词： Abiotic stress；abscisic acid；gene expression regulation；Tamarix hispida；WRKY transcription factor