抗植物病毒药剂毒氟磷抗病毒作用机制初步研究.pdfVIP

ThePreliminaryStudiesonActionMechanismof Anti-PlantViralAgent-Dufulin CHENZhuo SONGBao-an YANGSong BHADURYPinakiS HUDe-yu JINLin-hong XUEWei,ZENGSong CenterforResearchandDevelopmentofFineChemicals,KeyLaboratoryof GreenPesticideandBioengineering,MinistryofEducation,GuizhouUniversity, Guiyang550025,China Abstract:NicotianatabacumK326andTMVusedtostudyobject,theactionmechanismofanti-plant viralwascompletedbyusingmethodsofmolecularbiology,proteomicsandbio-chemistry.DIGEassayin dicatedthat40proteinspotswereinducedtoup-regulationintheleafinfectedbyTMVandtreatedby Dufulin.TheproteinspotswereidentifiedbyMSSpectrometryandrelatedwithdiseaseresistance,stress resistance,photosynthesis,cellwall,cytoskeletonandenergymetabolism.Theproteinsfromcytoplasm andchloroplastconsistedofabout45％ and37％ oftheallprotein.Theanalysisofproteinfunctionshown thatthedifferentialproteinwasrelatedwithinductionplantSAR;ThePRwasup-regulatedbyDufulinby usingtechniquesofsemi-quantityPCR,Real-timePCRandSDS.Itwasincreasedthatthedefense enzymeactivityandchlorophyllcontentinspecialperiodbyusingenzymebio-chemistrymethod.These resultsindicatedthatanti-viralactivitywasacquiredthroughactivationSAsignalpathway,increasing SARlevelandenhancingphytoalexin.Meanwhile,itwasfoundthatthelargesubunitandsmallsubunitof rubiscowereinducedbyDufulinforrectifyingdysfunctionofphotosynthesiscausedbyTMV. 抗植物病毒药剂毒氟磷抗病毒作用机制初步研究 陈卓 宋宝安 杨松 BHADURYPinakiS 胡德禹 金林红 薛伟 曾松 摘要：本文采用分子生物学、蛋白质组学研究和生物化学方法，以烟草NicotianatabacumK326和TMV 为对象，研究新型抗植物病毒药剂——毒氟磷抗植物病毒的作用机制。采用DIGE试验发现毒氟磷处理感 染TMV烟草叶片中约40个蛋白质点的表达上调，采用MS质谱对其进行鉴定，发现这些差异蛋白质点与 抗病性和抗逆性、光合作用、细胞骨架和能量代谢相关，其中与植物光合作用和抗病抗逆性相关的蛋白质 约占发现总的差异蛋白的35％和19％；通过蛋白的功能分析发现毒氟磷诱导差异蛋白与诱导植物SAR相 关；采用半定量PCR、Real-timePCR和SDS技术发现毒氟磷可诱导烟草PR-1a等