水稻pip2-1基因在rsv浸染过程中的功能研究-functional study of pip 2 - 1 gene in rice during rsv impregnation.docxVIP

AbstractEukaryotictranslationinitiationfactorisakindofimportantproteinsinvolvedinproteintranslationinitiationprocess.Itplayskeyrolesinthewholeproteinstranslationprocess.Alargenumberofstudieshaveshownthatmanytranslationinitiationfactorsarealsoinvolvedintheplantresistancepathway,besidesproteintranslationinitiationprocess.Eukaryotictranslationinitiationfactorhasalargeproteinfamily.ThePip2-1proteininthisstudyisoneofthefamilymembers.Inthiswork,weidentifiedinteractionbetweenPip2-1proteinandRSVP2proteinthroughtheYeasttwohybridandBimolecularfluorescencecomplementationexperiments.Subcellularlocalizationanalysisshowedthattheproteinlocatesonthecellmembrane.Whenweusedvirusinducedgenesilencing(VIGS)techniquetoinducePip2-1genesilencinginplant.Plantsshoweddwarf,leafmalformation,blossomanomaliesandlowseedsettingrate,whichindicatedthatPip2-1genewasessentialintheprocessofplantgrowthanddevelopment.TheresultsofReal-timePCRshowedthatthePip2-1geneexpressionlevelwasdown-regulatedinRSVinfectedtobaccoandriceplant,indicatingthatPip2-1mighthavecertainbiologicalfunctionintheprocessofRSVinfection.TheseresultsprovidecluesfortheRSVinfectionmechanismaswellasthehostresistancemechanism.InordertodofurtherinvestigationofPip2-1geneinthegrowthanddevelopementprocessofNicotianabenthamianaandriceaswellastherelationshipduringvirusinfectioncycle,weclonedNbPip2-1andOsPip2-1genesequencesfromNicotianabenthamianaandrice,andconstructedtheover-expressionvectorandtheinhibitionexpressionvectorofNbPip2-1andOsPip2-1.Byagrobacterium-mediatedtransformation,transgenicNicotianabenthamianaandriceplantswereobtained.Real-timePCRandWesternblotshowedthatthePip2-1genehadbeensuccessfullyintegratedintothegenomeoftransgenicplantsaswellasexpressedeffectively.Theseresultslaidanimportantfoundationforthelaterstudy,suchasthepHenotypeanalysis,functionidentificationandresistancereseachofthetransgenicplants.Keywords:Eukaryotictranslationinitiationfactor,Virusinfection,RSV,VIGS，Transgenosis目录摘要IAbstractII文献综述11水稻条纹病毒研究现状11.1水稻条纹叶枯病的症状表现11.