摘要
【目的】探究内质网胁迫(endoplasmic reticulum stress, ERS)引发的未折叠蛋白反应(unfolded protein response,UPR)与色氨酸代谢和植物抗病性的关系;明确UPR中的转录因子bZIP28和bZIP60在色氨酸代谢中的调控机制及对植物抗病性的影响。【方法】利用转录组测序和RT-qPCR分析内质网胁迫后,色氨酸代谢通路相关基因的表达;检测内质网胁迫对色氨酸代谢产物植保素-亚麻荠素(camalexin)和吲哚-3-乙酸(indole-3-acetic acid, IAA)合成的影响;检测病原菌侵染后bZIP28和bZIP60的表达及其过表达植株的抗病能力;通过酵母单杂交和双荧光素酶报告实验探究bZIP28/60对WRKY33的调控。【结果】UPR使色氨酸代谢基因表达上调,植保素Camalexin和IAA的水平升高;病原菌感染上调bZIP28和bZIP60的表达,且bZIP28和b ZIP60过表达植株的抗病性增强;bZIP28和b ZIP60间接调控WRKY33的转录,且WRKY33启动子的-364--566 bp区段为核心序列。【结论】内质网胁迫通过转录因子bZIP28和b ZIP60调控WRKY33的表达,进而上调色氨酸代谢基因的表达,提高Camalexin和IAA的水平,抵御病原菌的感染,缓解内质网胁迫。
【Objective】To investigate the relationship between endoplasmic reticulum stress(ERS)induced unfolded protein response(UPR),tryptophan metabolism,and plant disease resistance,and to clarify the regulatory mechanisms of the transcription factors bZIP28 and bZIP60 in tryptophan metabolism and their impact on plant disease resistance.【Method】RNA-seq and qRT-PCR were used to analyze the expressions of genes involved in the tryptophan metabolism pathway under ER stress conditions.The synthesis of tryptophan-derived metabolites,camalexin and IAA,was measured under ER stress.The expressions of bZIP28 and bZIP60 after pathogenic infection and the disease resistance of plants overexpressing these genes were assessed.Yeast one-hybrid assays and dual-luciferase reporter assays were conducted to examine the regulation of WRKY33 by bZIP28/60.【Result】ER stress triggered the UPR,leading to the up-regulation of tryptophan metabolic genes and increased levels of camalexin and IAA.Pathogenic infection up-regulated the expressions of bZIP28 and bZIP60.Plants overexpressing bZIP28/60 demonstrated enhanced resistance to disease.bZIP28/60 indirectly regulated the expression of WRKY33,with the core regulatory sequence located within the-364 to-566 bp region of the WRKY33 promoter.【Conclusion】ER stress modulates the expression of WRKY33 through the transcription factors bZIP28 and bZIP60,resulting in elevated levels of camalexin and IAA,thereby enhancing resistance to pathogen infection and alleviating ER stress.
作者
曹园园
周舒浩
张海荣
崔晓娜
CAO Yuan-yuan;ZHOU Shu-hao;ZHANG Hai-rong;CUI Xiao-na(College of Life Sciences,Henan Agricultural University,Zhengzhou 450046)
出处
《生物技术通报》
北大核心
2025年第8期155-164,共10页
Biotechnology Bulletin
基金
国家自然科学基金项目(31872660)。
