抗黄瓜花叶病毒RNAi载体的构建及烟草的转化

2021-09-24 20:08

AgriculturalScience&Technology,2010,11(5):69-72Copyright2010,InformationInstituteofHAAS.Allrightsreserved.?

PlantProtection

ConstructionofRNAiVectorWhichResistCucumberMosaicVirusandTransformationofTobacco

?ZHANGYu,ZHENGYin?ying,ZHAOFeng?yu,QIAOYa?hong,CUIBai?ming,XIANGBen?chun

KeyLaboratoryofAgricultrureBiotechnologyofShiheziUniversity/CollegeofLifeSciences,ShiheziUniversity,Shihezi832003

Abstract [Objective]AimedtoconstructRNAivectorresistanttocucumbermosaicvirusandtransferredthisvectorintotobacco.[Method]RT?PCRmethodwasusedtoamplifycucumbermosaicvirusNS04andprocessRNA2genesequenoftomatoisolates.TheanalysisresultsofphylogenetictreedemonstratedthatthesequenceinRNA2encodedCMV?2ahad98.0%and96.5%homologywithnucleotideandaminoacidof

,China.ThereplicasefragmentinCMVRAN2genewastakenastargetsequencetoconstructpBi35SCR2eukary?DQ412731isolateofZhejiang

oticexpressionvector,thentheexpressionvectorwasidentified.Throughagrobacterium?mediatedmethod,theexpressionvectorwastrans?ferredintotabaccoandPCRmethodwasusedtocheckthetransfer.ThePCRresultsdemonstratedthattheexperimenthadsuccessfullycon?structeukaryoticexpressionvectorofpBi35SCR2andtheexpressionvectorwassuccessfullytransferredintotabacco.[Conclusion]Theob?tainedtransgenictobaccocouldbeusedaschallengetestmaterialinfollowingexperimentandprovidedfoundationforstudyingprocessingtomatoresistcucumbermosaicvirus.Keywords Cucumbermosaicvirus;Homology;RNAivector;Tobacco;Transformation

  Cucumbermosaicvirus(CMV)isanimportantpathogentocauseyieldreductionandevennoharvestofavarietyofeconomiccrops,butthetraditionalanti?virusbreeding,tissuedetoxificationandpesticidepreventionmethodscauseenvi?

[1]

ronmentalpollution,andtimeconsuming.Inrecentyears,thediscoveredphenomenonofgenesilencingisacommonphenomenonexistedintheprocessofbiologicallife,whichisanimportantwayoftheregulationofgeneexpression.ThegenesilencingofGM?specificcanbetransmittedbygraftingfromsilentstocktonon?silentscion,thenthephenomenonofthetransferringofthiskindofgenesilencingsignalfromonecelloratissuetoanothercellortissueisdefinedassystemic

[2-5]

RNAi(RNAinterference).RNAihassomeadvantageswhicharedeficientinthetraditionalgeneticresearchmeth?ods,ithasprovidedanewwaytostudyplantgenefunctionandgeneexpressionregulation,andthefunctionaldeletantcanbecreatedthroughtheselectivelyinhabitationsoftheex?

[6]

pressionofspecificgenes.Combinedwiththespecific,effi?

,RNAitechnologyhasacientandsystematicRNAisequence

,ithasbeendemonstrateditsuniqueroleparticularlyspecialrole

inthepreventionandtreatmentofviruses,wormsandharmful

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