小麦矮缩病毒实时荧光定量pcr体系在小麦和异沙叶蝉中的建立与应用

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1、万方数据AbstractWheatdwarfvirus(WDV)，transmittedbyleathopper(PsammotettixstriatusL．)inapersistentcirculativemannegbelongstothegenusMastrevirus，familyGeminiviridae．WDVandleafhopperorwheatinteractedgeneexpressionplaysanimportantroleforspreadofwheatdwarfdisease．IthasbeenprovedthatGeminiviridaecoatproteina

2、ctasanauxiliaryforMP,eitheractivelyorbyprotectingthegenomewhenthevirusmovescelltocellthroughplasmodesmata．TheCPproteinofWDVhasbeeninteractedwithleafhoppetThepurposeofthisstudyistoanalysisWDVgeneexpressionpatterninwheatandleafhopperusingreal-timequantitativePCR(RT-qPCR)．(1)WebuilttheRT-qPCRprotocoli

3、nregardtostudygeneexpressioninthewheat，using5housekeepinggenesTEF一1-0l，GAPDH，18SrRNA，UBCand28SrRNAasthecandidatereferencegenesGenesexpressionstabilitywasassessedinthestemandleaftissueofwheatusingGenorm，NormFinderandBestkeepersoffwares．Inthestemtissue．TEF一1—0【rankedasthemoststableandthe28SrRNAisthel

4、eaststableone．Whereas，inleaftissuegenestabiltywasvaringinallthethreesoftwareusedforanalysisgeneexpression．TEF一1一olfollowedby18SrRNA，GAPDH，28SrRNAandUBCshowedmorestabilityinGenormanalyticalsoftware．GAPDHwasprovedmorestableascompareto18SrRNA，TEF·I一0L，UBCand28SrRNAinNormFindersoftware．Ontheotherhand，B

5、estkeepersoftwareshowed18SrRNAasmorestableinleavestissuesascomparetootherfourgenes．HoweveLTEF·1-0tandGAPDHweremoresatbleinbothtissuesandtheoptimalnumberofreferencegenesfornormalizationwasatIeastfive．(2)ThisstudyanalyzedtherelativeexpressionofWDV-CPgeneinthestemandleafofwheatbytheRT-qPCRsystemwebuil

6、t．TherelativequantitativeofWDV-CPgeneinthestemwashigherthantheleaf．ThismightbeastheresultoftheenteringofWDVinthewheatphloembytheleafhopperandthenmoveinthephloemuptodown．(3)TheRT-qPCRsystemwasbuiltintheleafhopperusing6housekeepinggenes(RPS23e，UBC，Beta-TUB，GAPDH，18SrRNAandACT2)whichwerethefirsttimese

7、quencedfromtheleathopper．OurresultsshowedthatRPS23ewasmoststableandACT2waslessstableascomparetoUBC，Beta-TUB，GAPDHand18SrRNAinleafhopperusingeitheranalyticalsoftware．However，themoststablecombinationwasBeta—T