大丽轮枝菌蛋白激发子PevD1激活本生烟促分裂原活化蛋白激酶MAPK  

Verticillium dahliae Elicitor PevD1 Activates MAPKs in Nicotiana benthamiana

作　　者：贾丰莲 李泽 梁颖博 李广悦[1] 杨秀芬[1] JIA Feng-lian;LI Ze;LIANG Ying-bo;LI Guang-yue;YANG Xiu-fen(The State Key Laboratory for Biology of Plant Disease and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Science,Beijing 100081)

机构地区：[1]中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京100081

出　　处：《生物技术通报》2020年第10期15-24,共10页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31772151);国家重点研发计划项目(2017YFD0201100)。

摘　　要：激活植物免疫系统、提高植物自身抗性是植物病害绿色防控的重要途径之一。促分裂原活化蛋白激酶(MAPK)是植物免疫系统的重要防御信号传导途径。为了探讨大丽轮枝菌蛋白激发子PevD1诱导植物广谱抗病性的分子机制,前期利用转录组测序(RNA-Seq)技术获得了本生烟响应PevD1诱导的差异表达基因,其中大量基因显著富集在MAPK通路上。本研究对这些差异表达基因进行了功能分类并进一步分析,发现这些基因涉及的功能十分广泛,包括参与植物识别的蛋白激酶(LRR-RLK)、调控基因表达的转录因子家族(WRKY和ERF)、与抗病相关的几丁质酶基因、参与钙离子信号传递的钙调蛋白(Calmodulin)、调控活性氧(Reactive oxygen species,ROS)产生的呼吸爆发氧化酶(Rboh)和清除ROS的过氧化氢酶(Catalase)等。从富集在MAPK通路中的差异表达基因中选出10个基因进行了qPCR验证,转录表达模式与转录组测序结果一致。用特异性磷酸化位点抗体杂交技术验证了PevD1激活了本生烟的MAPK,即水杨酸诱导的蛋白激酶SIPK和伤诱导的WIPK被激活。Plant resistance enhancement via activating plant immune system is one of the important approaches in green management system of plant diseases and pests.Mitogen activated protein kinase(MAPK)cascade is a vital signal transduction pathway in plant innate immune system.In order to investigate the molecular mechanism of Verticillium dahliae elicitor PevD1 inducing broad-spectrum disease resistance,transcriptome sequencing(RNA-seq)data from Nicotiana benthamiana response to PevD1 was obtained and a large number of differential gene expression(DGEs)were significantly enriched in the MAPK pathway.In this paper,DGEs enriched in the MAPK pathway were functionally classified and further analyzed.The involved functions by these DGEs were broad,including protein kinases(LRR-RLK)associated with plant recognition,ERF and WRKY transcription factor family for the regulation of gene expression,chitinase gene involved in disease resistance,calmodulin participated in calcium signaling,breathing outbreak oxidase(Rboh)regulating reactive oxygen species(ROS)and catalase(CAT)clearing ROS,etc.Expression patterns of 10 genes from the DGEs enriched in MAPK pathway were verified by qPCR and it was consistent with the RNA-Seq result.The antibody hybridization technique of specific phosphorylation sites was used to confirm that PevD1activated MAPK in N.benthamiana,i.e.,salicylic acid-induced protein kinase SIPK and wound-induced WIPK were activated by PevD1.

关 键 词：蛋白激发子PevD1 转录组测序 MAPK SIPK WIPK 

分 类 号：S432.1[农业科学—植物病理学]