机构地区:[1]浙江大学农业与生物技术学院,杭州310058 [2]浙江大学海南研究院,海南三亚572025
出 处:《中国农业科学》2022年第16期3144-3154,共11页Scientia Agricultura Sinica
基 金:浙江省自然科学基金杰青项目(LR19C150001);浙江省重点研发计划(2021C02040)。
摘 要:【背景】近年来,我国番茄生产中面临的不良气候环境导致露地和设施栽培中番茄病害日益严重。由丁香假单胞菌番茄致病变种(Pseudomonas syringae pv.tomato,Pst)所引发的番茄细菌性叶斑病频发,严重影响了番茄的产量、品质。糖是植物体内重要的信号分子与营养物质,在植物遭受病原菌侵染时,糖既可以作为信号参与调控植物抗病性,也可以作为主要碳源为免疫反应提供能量。STP(sugar transport protein)家族是一类糖转运蛋白家族,在植物生长发育过程与病原防御中起着重要作用。【目的】明确番茄中STP家族是否参与调控植株对细菌性叶斑病的抗性。【方法】以番茄CR品种(Solanum lycopersicum cv.Condine Red)为材料,通过接种Pst DC3000,明确STP基因家族对Pst DC3000的响应。构建SlSTP2的突变材料与过表达材料,并进行鉴定、繁种。在野生型和SlSTP2基因突变体、过表达植株上接种Pst DC3000,观察比较叶片发病情况、台盼蓝染色显示的死细胞数量,检测叶片菌落数(colony-forming units,CFU)和光系统Ⅱ实际光化学效率,明确SlSTP2在植株防御细菌性叶斑病中的作用。为探究SlSTP2防御Pst DC3000的内在分子机制,通过双分子荧光互补(bimolecular fluorescent complimentary,BiFC)试验筛选互作蛋白,构建编码该蛋白的基因突变与过表达材料,并接种Pst DC3000明确其在植株防御细菌性叶斑病中的作用。【结果】在番茄植株上接种Pst DC3000后,SlSTP1和SlSTP2表达上调,选取上调最显著的SlSTP2作为后续研究对象,构建其突变材料与过表达材料。在番茄野生型和Slstp2突变植株、OE:SlSTP2过表达植株上接种Pst DC3000,相比野生型植株,Slstp2植株细菌性叶斑病的发病情况更严重,叶片CFU与台盼蓝染色显示出的死细胞数均更多,光系统Ⅱ实际光化学效率下降,OE:SlSTP2植株则相反。通过BiFC试验发现,SlSTP2与G蛋白β亚基SlAGB1互作。接种Pst DC3000后,Sl【Background】In recent years,tomato diseases occur frequently in open field and protected cultivation due to the suboptimal environmental conditions.Tomato bacterial leaf spot caused by Pseudomonas syringae pv.tomato(Pst)is a common bacterial disease,which severely affects the yield and quality of tomato.Sugar is an important signaling molecule as well as nutrient substance in plants.When plants are subjected to pathogen attack,sugar not only functions as a signal to regulate the plant defense,but also provides energy for immune responses as a main carbon source.Sugar transport protein(STP)family is responsible for the sugar transport,and plays an important role in plant growth,development and defense.【Objective】The objective of this study is to clarify whether STP family is involved in the regulation of plant defense against bacterial leaf spot.【Method】CR(Solanum lycopersicum cv.Condine Red)was used as the wild-type(WT)background in this study.The responses of STP gene family to Pst DC3000 were determined by Pst DC3000 inoculation.The SlSTP2,which was up-regulated most significantly after Pst DC3000 inoculation,was selected for further investigation.The Slstp2 homozygous mutant and overexpression materials were generated.To explore the role of SlSTP2 in plant defense against bacterial leaf spot,WT and Slstp2 mutants,OE:SlSTP2 plants were inoculated with Pst DC3000,then the disease symptoms were assessed by disease-associated cell death and bacterial growth as well as the photochemical efficiency.In order to explore the potential mechanism of SlSTP2 in the disease resistance,the interacting proteins were screened by bimolecular fluorescence complementary(BiFC)assay.The mutant and overexpression materials of candidate interacting protein were generated and inoculated with Pst DC3000 to investigate its role in plant defense against bacterial leaf spot.【Result】After inoculation with Pst DC3000 on tomato plants,the expressions of SlSTP1 and SlSTP2were up-regulated.SlSTP2 was up-regulated most significa
关 键 词:番茄 糖转运蛋白 SlSTP2 细菌性叶斑病 抗病性
分 类 号:S436.412.1[农业科学—农业昆虫与害虫防治]
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