毛白杨转录因子PtWRKY01调控靶标基因表达的分子机制

项目名称： 毛白杨转录因子PtWRKY01调控靶标基因表达的分子机制

项目编号： No.31270698

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 林善枝

作者单位： 北京林业大学

项目金额： 82万元

中文摘要： 杨树叶锈病造成巨大经济损失，已引起人们高度关注。本项目以前期研究证实与毛白杨抗锈病密切相关转录因子PtWRKY01为目标，采用染色质免疫沉淀结合高通量测序(ChIP-seq)技术，获得全基因组内与转录因子PtWRKY01互作的DNA序列，进而通过计算分析确定PtWRKY01所调控的靶标基因及其转录表达所涉及的信号通路。同时，对前期已获得未转基因、RNA干扰后以及过表达PtWRKY01毛白杨进行锈病病原菌活体接种实验，开展与抗病密切相关代谢物的测定，并对病程相关蛋白基因、抗病信号转导分子基因以及PtWRKY01所调控靶标基因的转录表达进行检测，从中探讨PtWRKY01在毛白杨抗病免疫系统中的作用机理，进而从功能基因组学、转录组学、代谢组学以及信号通路等层面阐明毛白杨转录因子PtWRKY01调控靶标基因表达的分子机制，这可为PtWRKY01应用于改良某些有经济价值的农作物与林木奠定重要基础。

中文关键词： 毛白杨抗病性；WRKY转录因子PtWRY01；ChIP-Seq技术；转录组测序；基因表达调控机制

英文摘要： Recently, much attention has been focused on the problem about the growth and productivity of the poplars has adversely affected by leaf rust. Our previous studies have demonstrated that WRKY-type transcription factor PtWRKY01 from Populus tomentosa, act as one key defense-related regulator, directly participated in the transcriptional regulation of defense responses against pathogens. In the present study, the anti-PtWRKY01 polyclonal antibody with high specificity, obtained by immunized rabbit with the previously purified fusion protein His-PtWRKY01, can be used for the chromatin immunoprecipitation (ChIP) analysis of PtWRKY01 transcription factor, and the potential target DNAs specifically binding to PtWRKY01 could be obtained on the genome scale by the ChIP coupled with high-throughput sequencing (ChIP-Seq) technique. Through combination with gene ontology analysis and pathway analysis, the transcription factor PtWRKY01-regulated downstream target genes were identified, and the PtWRKY01-mediated possible defense signaling pathways were revealed. Additionally, to elucidate the role of PtWRKY01 transcription factor involved in the regulation of defense signaling pathways during the responses of P. tomentosa to pathogen-resistance, after the previously obtained transgenic P. tomentosa plants of over-expressing

英文关键词： Populus tomentosa disease resistance；WRKY transcription factor PtWRKY01；ChIP-Seq technique；Illumina transcriptomic sequencing；Regulatory mechanism of gene expression