细胞壁转化酶调控番茄响应灰霉菌侵染的转录组分析  

Transcriptome Analysis of CWIN-mediated Tomato Response to the Infection of Botrytis cinerea

作　　者：付兰平 辛曙丽 刘永华 朱国鹏[1,3] FU Lanping;XIN Shuli;LIU Yonghua;ZHU Guopeng(School of Horticulture,Hainan University/Key Laboratory for Quality Regulation of Tropical Horticultural Crops of Hainan Province,Haikou,Hainan 570228,China;Agricultural Service Center of Baoting,Baoting,Hainan 572316,China;Sanya Nanfan Research Institute,Hainan University,Sanya,Hainan 572022,China)

机构地区：[1]海南大学园艺学院/海南省热带园艺作物品质调控重点实验室,海南海口570228 [2]保亭黎族苗族自治县农业服务中心,海南保亭572316 [3]海南大学三亚南繁研究院,海南三亚572022

出　　处：《热带作物学报》2024年第6期1139-1156,共18页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.31760579);海南省研究生创新课题(No.Qhys2021-250)。

摘　　要：由死体营养型病菌(necrotrophic pathogen)灰葡萄孢菌(Botrytis cinerea)侵染引起的灰霉病是番茄生产上的常见病害,可导致番茄大面积减产甚至绝收。蔗糖分解代谢在植物抗病中发挥着重要作用,不仅可为植物防御反应提供碳骨架和能量,还可通过信号途径调控抗病基因的表达。研究表明,分解蔗糖的细胞壁转化酶(cell wall invertase,CWIN)可增强植物对死体营养型病害的抗性。然而,目前尚无CWIN对番茄灰霉病抗性影响的相关研究。本研究以CWIN活性上调的转基因番茄(RNAi/R)及野生型番茄(WT/W)为材料,对其叶片进行灰霉菌(Bc)离体接种,研究CWIN对番茄灰霉病抗性的影响,同时对接种后12、60 h的叶片进行取样,通过转录组测序初步阐明CWIN调控番茄灰霉病抗性的分子机制。结果表明:(1)提高番茄CWIN活性可增强番茄叶片对灰霉病菌的抗性。(2)KEGG注释表明,接种12 h的DEGs(W-Bc-12 h-vs-R-Bc-12 h)共获得5个显著性富集通路,包括次生代谢物生物合成(biosynthesis of secondary metabolites)、代谢途径(metabolic pathways)、DNA复制(DNA replication)、淀粉和蔗糖代谢(starch and sucrose metabolism)以及甾族化合物生物合成(steroid biosynthesis);接种60 h的DEGs(W-Bc-60 h-vs-R-Bc-60 h)则未发现显著性富集通路,表明接种早期是CWIN调控番茄抗病性的关键时期。(3)植物-病原菌互作图分析表明,参与超敏反应和防御相关基因诱导的富含亮氨酸重复(LRR)受体类丝氨酸/苏氨酸蛋白激酶基因FLS2和热激蛋白基因HSP90在接种后的RNAi叶片中上调,其可能是CWIN增强番茄抗病性机制中的重要基因。(4)植物激素信号通路和MapMan作图分析表明,接种后RNAi番茄的抗病激素茉莉酸(JA)和乙烯(ET)信号途径上调,同时水杨酸(SA)信号途径减弱,表明其可能共同作用提高RNAi番茄的抗病性。此外,接种后RNAi番茄叶片的生长促进激素生长素(IAA)和细胞分裂素(CTK)信号途径增强,Grey mold is a common disease in tomato production caused by Botrytis cinerea,a necrotrophic pathogen,which often leads to dramatic reduction of tomato yield.Sucrose catabolism plays an important role in plant defense against pathogen infection by providing carbon skeleton and energy for plant defense responses and/or regulating the expression of defense-related genes through signaling pathway.Previous studies have shown that cell wall invertase(CWIN),a kind of sucrose-degrading enzyme,can enhance plant resistance to several necrotrophic pathogens.However,no research has been conducted to study the role of CWIN in tomato resistance to B.cinerea.In this study,wild type tomato(W)and its transgenic line(R)with elevated CWIN activity were used as materials to study the effect of CWIN on tomato resistance to B.cinerea(Bc)via in vitro inoculation.In addition,inoculated leaves were sampled 12 h and 60 h post inoculation(hpi)for RNA-Seq to elucidate possible molecular mechanisms underlying the regulation of CWIN to tomato resistance against B.cinerea.The results are as follows:(1)Elevated CWIN activity enhanced tomato resistance to B.cinerea;(2)KEGG annotation showed that DEGs(W-Bc-12 h-vs-R-Bc-12 h)from 12 hpi were significantly enriched in five pathways,including biosynthesis of secondary metabolites,metabolic pathways,DNA replication,starch and sucrose metabolism,and steroid biosynthesis;No significant enrichment pathway was found for DEGs(W-Bc-60 h-vs-R-Bc-60 h)from 60 hpi.(3)By mapping DEGs to plant-pathogen interaction pathway,it was revealed that the LRR-receptor serine/threonine-like kinase gene FLS2 and heat shock protein gene HSP90 involved in hypersensitive response and defense-related gene induction were up-regulated in RNAi leaves after inoculation,indicating the two genes may participate in the regulation of CWIN to tomato resistance to B.cinerea.(4)The analysis of plant hormone signal transduction pathways and MapMan mapping showed that the signal pathway of jasmonic acid(JA)and ethylene(ET)was enhanced in

关 键 词：细胞壁转化酶 灰霉菌(Botrytis cinerea) 番茄 转录组测序 抗病基因 

分 类 号：S436.412.13[农业科学—农业昆虫与害虫防治]