Virus-induced gene-silencing in wheat spikes and grains and its application in functional analysis of HMW-GS-encoding genes 英文参考文献.docVIP

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Virus-induced gene-silencing in wheat spikes and grains and its application in functional analysis of HMW-GS-encoding genes 英文参考文献.doc

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Virus-induced gene-silencing in wheat spikes and grains and its application in functional analysis of HMW-GS-encoding genes 英文参考文献

Maetal.BMCPlantBiology2012,12:141 /1471-2229/12/141 RESEARCH ARTICLE OpenAccess Virus-inducedgene-silencinginwheatspikesand grainsanditsapplicationinfunctionalanalysisof HMW-GS-encodinggenes MengMa1,2,YanYan1,2,LiHuang3,MingshunChen4andHuixianZhao1,2* Abstract Background:TheBarleystripemosaicvirus(BSMV)-basedvectorhasbeendevelopedandusedforgenesilencingin barleyandwheatseedlingstoassessgenefunctionsinpathogen-orinsect-resistance,butconditionsforgene silencinginspikesandgrainshavenotbeenevaluated.Inthisstudy,weexploredthefeasibilityofusingBSMVfor genesilencinginwheatspikesorgrains. Results:ApparentphotobleachingonthespikesinfectedwithBSMV:PDSatheadingstagewasobservedafter13days postinoculation(dpi),andpersisteduntil30dpi,whilethespikesinoculatedwithBSMV:00remainedgreenduringthe sameperiod.GrainsofBSMV:PDSinfectedspikesalsoexhibitedphotobleaching.Molecularanalysisindicatedthat photobleachedspikesorgrainsresultedfromthereductionofendogenousPDStranscriptabundances,suggestingthat BSMV:PDSwasabletoinducePDSsilencinginwheatspikesandgrains.Inoculationontowheatspikesfromheadingto floweringstagewasoptimalforefficientsilencingofPDSinwheatspikes.Furthermore,weusedtheBSMV-basedsystem toreducethetranscriptlevelof1Bx14,ageneencodingforHigh-molecular-weightgluteninsubunit1Bx14(HMW-GS 1Bx14),by97%inthegrainsoftheBSMV:1Bx14infectedspikesat15dpi,comparedwiththatinBSMV:00infectedspikes, andthereductionpersisteduntilatleast25dpi.TheamountoftheHMW-GS1Bx14wasalsodetectablydecreased.The percentageofgluteninmacropolymericproteinsintotalproteinswassignificantlyreducedinthegrainsof1Bx14-silenced plantsascomparedwiththatinthegrainsofBSMV:00infectedcontrolplants,indicatingthatHMW-GS1Bx14isoneof majorcomponentsparticipatingintheformationofgluteninmacropolymersinwheatgrains. Conclusion:Thisisoneof thefirstreportsofsuccessfulapplicationofBSMV-basedvirus-induced-gene-silencing(VIGS) forgeneknockdowninwheatspikesandgrainsanditsapplicationinfunctionalanalysisofthe1Bx14gene.The establishedBSMV-VIG