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