单链DNA结合蛋白WHIRLY1转录及表观遗传调控植物衰老和细胞死亡的研究

项目名称： 单链DNA结合蛋白WHIRLY1转录及表观遗传调控植物衰老和细胞死亡的研究

项目编号： No.31470383

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 植物学

项目作者： 缪颖

作者单位： 福建农林大学

项目金额： 85万元

中文摘要： 本项目基于已证明单链DNA结合蛋白WHIRLY1是WRKY的上游调控因子，结合在WRKY53启动子上，激活或抑制WRKY基因的表达而调控衰老与细胞死亡的基础，利用WHIRLY1突变体通过荧光原位杂交和荧光标记技术、染色质免疫共沉淀序列分析和PCR分析以及实时定量PCR技术，寻找WHIRLY1参与调控下游基因转录的证据，即WHIRLY1是否通过改变染色质构象，或参与直接招募RNA聚合酶II，或直接和间接参与调控组蛋白的修饰等转录及其表观遗传调控事件，进而调控下游衰老相关基因WRKY53和细胞死亡相关基因WRKY33的表达，探明WHIRLY1转录及表观遗传调控下游基因表达的分子机制；并通过对不同发育时期和不同衰老和细胞死亡诱导处理材料的研究，阐明单链DNA结合蛋白调控植物细胞衰老与细胞死亡启动与进程的分子机理。这将是植物衰老调控理论的补充与深化，也为有效控制作物产量、品质和保鲜等提供理论依据。

中文关键词： 单链DNA结合蛋白；转录调控；表观遗传调控；衰老与细胞死亡；拟南芥

英文摘要： Senescence is the last phase of leaf development finally leading to death of the tissue. This phase of development is characterized by remobilization of nutrients and in crop plants limits yield and energy transmission. It has been shown that transcription factors of the WRKY and NAC families have prominent roles in controlling the process. Our previous research has demonstrated that single stranded DNA binding proteins, WHIRLY constitute another family of transcription factors playing key role in senescence as well as in cell death processes during biotic or abiotic stresses whereby WHIRLY1 acts upstream of WRKY transcription factors. WHIRLY1 binds to the promoter of WRKY53 and activates or represses downstream WRKY gene expression and senescence events. The major goal of the project is to elucidate the molecular mechanism of the upstream regulation of WRKY transcription factors by WHIRLY1 using fluorescence in situ hybridization，ChIP sequencing， quantitative PCR and quantitative reverse transcription PCR assay. In addition, chromatin remodeling, recruitment of RNA polymerase II and epigenetic histone modification will be investigated using WHIRLY1 mutants. It aims to elucidate the molecular mechanism underlining Whirly1 actions on the downstream WRKY genes expression at the transcriptional and epigenetic levels. Furthermore, the developmental cue and environmental condition including darkness and H2O2 treatments will be included to decipher the molecular mechanism of Whirly1 upstream controls plant senescence and cell death.

英文关键词： single-stranded-DNA binding protein;transcriptinal control;epigenetical control;senescence and cell death;Arabidopsis