转录因子GhWRKY41促进水杨酸合成增强棉花对黄萎病菌的抗性  

Transcription factor GhWRKY41 enhances cotton resistance to Verticillium dahliae by promoting SA synthesis

作　　者：肖胜华 董贤镘 彭鑫 李安子 闭兆福 廖铭静 黄礼豪 管倩倩 胡琴 朱龙付[2,3] XIAO Sheng-Hua;DONG Xian-Man;PENG Xin;LI An-Zi;BI Zhao-Fu;LIAO Ming-Jing;HUANG Li-Hao;GUAN Qian-Qian;HU Qin;ZHU Long-Fu(State Key Laboratory of Conservation and Utilization of Agro-Biological Resources in Subtropical Region/College of Agriculture,Guangxi University,Nanning 530004,Guangxi,China;State Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070,Hubei,China;Hubei Hongshan Laboratory,Huazhong Agricultural University,Wuhan 430070,Hubei,China)

机构地区：[1]广西大学农学院/亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004 [2]华中农业大学/作物遗传改良国家重点实验室,湖北武汉430070 [3]华中农业大学/湖北洪山实验室,湖北武汉430070

出　　处：《作物学报》2024年第10期2447-2457,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(32301880,32230076);湖北洪山实验室基金项目(2021hszd006)资助。

摘　　要：棉花是我国最重要的经济作物之一,但其产量和品质受黄萎病菌危害而大幅度下降。挖掘棉花黄萎病抗性相关基因并解析其分子机制,对加快棉花抗黄萎病育种进程具有重要意义。前期研究鉴定到一个在多个抗病棉花品种中均受黄萎病菌诱导显著上调表达的WRKY基因GhWRKY41,该基因通过激活苯丙烷代谢增强棉花对黄萎病菌的抗性。本研究进一步分析了GhWRKY41在不同激素处理下的诱导表达模式,利用病毒介导的基因沉默(VIGS)技术验证了GhWRKY41的抗病功能,并利用前期已创制的GhWRKY41转基因棉花株系‘Jin668’测定了其内源植物抗病激素含量。结果显示,GhWRKY41受水杨酸(SA)、Me-JA和H_(2)O_(2)诱导均显著上调表达;GhWRKY41被沉默后可削弱棉花对黄萎病菌的抗性;SA含量在GhWRKY41超表达棉花中显著增加,而在GhWRKY41干涉棉花中则明显减少。RT-qPCR分析显示,SA合成基因GhSID2及SA信号转导基因GhNPR1、GhPR1、GhPR5的表达水平在超表达植株中明显上调,而在干涉植株中则显著下降。ChIP-qPCR和双荧光素酶报告基因试验结果表明,GhWRKY41可结合并激活GhSID2、GhPR1和GhPR5的表达。此外,外施SA可明显提高棉花的黄萎病抗性。综上表明,GhWRKY41可通过促进棉花中内源植物激素SA的合成来增强植株的黄萎病抗性,这一结果完善了GhWRKY41在棉花抵御黄萎病菌过程中的生物学功能,为未来利用黄萎病抗性基因创制棉花抗病材料提供了理论基础。Cotton is an important commercial crop,and its yield and quality are severely affected by Verticillium dahliae.Identifying cotton resistance genes to Verticillium dahliae and exploring the underlying molecular mechanisms is of great significance for accelerating the breeding process of cotton resistant to Verticillium wilt.In a previous study,the WRKY gene GhWRKY41 was identified as being induced by Verticillium dahliae in multiple resistant cotton varieties,enhancing cotton resistance through the activation of phenylpropanoid metabolism.This study further analyzed the expression patterns of GhWRKY41 under different hormone treatments,validated its disease resistance function in the upland cotton variety‘Jin668’using a virus-induced gene silencing(VIGS)assay,and measured endogenous hormone content.The results demonstrated that GhWRKY41 was significantly up-regulated by SA,Me-JA,and H_(2)O_(2).Silencing GhWRKY41 weakened cotton resistance to Verticillium dahliae,while overexpression of GhWRKY41 led to a marked increase in SA content,and its RNAi lines showed a decrease in SA content.RT-qPCR results revealed that the expression levels of the SA biosynthesis gene GhSID2 and the SA signal transduction genes GhNPR1,GhPR1,and GhPR5 were significantly up-regulated in GhWRKY41-overexpression plants but decreased in GhWRKY41-RNAi plants.ChIP-qPCR and luciferase reporter gene assays showed that GhWRKY41 binds to and activates the expression of GhSID2,GhPR1,and GhPR5.Additionally,external spraying of SA significantly enhanced cotton resistance to Verticillium dahliae.In summary,GhWRKY41 enhances cotton resistance to Verticillium dahliae by promoting SA synthesis.This study elucidates the biological function of GhWRKY41 in cotton resistance to Verticillium dahliae and provides a theoretical basis for developing cotton varieties with improved resistance.

关 键 词：棉花 黄萎病菌 GhWRKY41 水杨酸 

分 类 号：S562[农业科学—作物学]