CsPUB54与PmRXLR1互作负向调控黄瓜对疫病的抗性  

CsPUB54 Negatively Regulates Cucumber Resistance to Phytophthora Melonis by Interacting with PmRXLR1 Effector

作　　者：王瑞[1,2] 吴红霏 张长远[1] 曹海顺 谭德龙[1] 郭金菊 王云龙 王茹芳 袁余 吴廷全[1,2] WANG Rui;WU Hongfei;ZHANG Changyuan;CAO Haishun;TANG Delong;GUO Jinju;WANG Yunlong;WANG Rufang;YUAN Yu;WU Tingquan(Institute of Facility Agriculture,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Vegetable Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China)

机构地区：[1]广东省农业科学院设施农业研究所,广州510640 [2]广东省农业科学院蔬菜研究所,广州510640 [3]仲恺农业工程学院,广州510225

出　　处：《园艺学报》2025年第3期591-602,共12页Acta Horticulturae Sinica

基　　金：国家自然科学基金面上项目(32272713);广州市科技计划项目(重点研发计划)(2024B03J1217);广东省自然科学基金项目(2019A515011310)。

摘　　要：以高感疫病的黄瓜B80和瓜类疫霉菌(Phytophthoramelonis)为材料,从B80黄瓜中克隆了1个含有U-box功能域的PUB蛋白家族基因CsPUB54,从P.melonis中克隆到1个RXLR效应蛋白基因,命名为PmRXLR1。B80接种P.melonis24h的转录组和q RT-PCR的试验结果均显示,P.melonis对B80黄瓜的侵染能够诱导CsPUB54和PmRXLR1表达量的快速提升。通过基因瞬时沉默方法将CsPUB54进行沉默,能够增加黄瓜子叶对P.melonis的抗性。另外,酵母双杂交和Pull-down的试验结果证实CsPUB54与PmRXLR1在体内和体外均可互作。利用碱基突变的方法将PmRXLR1的W和Y功能域分别进行突变,产生7个突变体。这些突变体分别与CsPUB54的互作检测显示,PmRXLR1中的W和Y功能域是PmRXLR1和CsPUB54互作所必需的,而W功能域中的异亮氨酸(I)是互作必需的关键氨基酸。CsPUB54与已发表的植物免疫负调控PUB蛋白进行遗传进化分析显示,CsPUB54与拟南芥AtPUB22、AtPUB23的遗传变异率较小,亲缘关系最近。这些研究数据表明CsPUB54能够通过与PmRXLR1互作负向调控黄瓜对瓜类疫霉的抗性。B80 cucumber with highly susceptible to Phytophthora blight,and Phytophthora melonis(P.melonis)were used as experimental materials.The gene that encodes a PUB protein containing the U-box domain from B80,named CsPUB54,and a RXLR effector protein gene,named PmRXLR1 from P.mellonis were cloned.Experimental results of transcriptome and qRT-PCR of cucumber leaves inoculated with P.mellonis for 24 hours both showed that the infection could induce rapid increase of gene expression levels of CsPUB54 and PmRXLR1.CsPUB54 gene silencing through transient gene silencing method could increase the resistance of cucumber cotyledons to P.melonis.In addition,the results of yeast two hybrid and Pull-down experiments confirmed that CsPUB54 could interact with PmRXLR1 both in vivo and in vitro.Using the method of base mutation,the W and Y functional domains of PmRXLR1 were mutated respectively,resulting in 7 mutants.The interaction detection between these mutants and CsPUB54 showed that the W and Y functional domains were necessary for the interaction between PmRXLR1 and CsPUB54,and the isoleucine(I)in the W functional domain was a key amino acid required for the interaction.Genetic evolution analysis of CsPUB54 and PUB proteins related to plant immune negative regulatory that have been published showed that CsPUB54 had a low genetic variation rate and the closest relationship with AtPUB22 and AtPUB23 in Arabidopsis thaliana.These research data suggest that CsPUB54 can negatively regulate cucumber resistance to P.melonis through interaction with PmRXLR1.

关 键 词：黄瓜 疫病 瓜类疫霉 蛋白互作 负向调控 抗性 

分 类 号：S642.2[农业科学—蔬菜学]