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1、TestingInsecticidalActivityofNovelChemicallySynthesizedsiRNAagainstPlutellaxylostellaunderLaboratoryandFieldConditionsLiangGong,YongChen,ZhenHu,MeiyingHu*KeyLaboratoryofPesticideandChemicalBiology,CollegeofNaturalResourcesandEnvironment,SouthChinaAgriculturalUniversity,Guangzhou,China
2、AbstractBackground:Overthelast60years,syntheticchemicalpesticideshaveservedasamaintacticinthefieldofcropprotection,buttheiravailabilityisnowdecliningasaresultofthedevelopmentofinsectresistance.Therefore,alternativepestmanagementagentsareneeded.However,thedemonstrationofRNAigenesilenci
3、ngininsectsanditssuccessfulusageindisruptingtheexpressionofvitalgenesopenedadoortothedevelopmentofavarietyofnovel,environmentallysoundapproachesforinsectpestmanagement.Methodology/PrincipalFindings:SixsmallinterferingRNAs(siRNAs)werechemicallysynthesizedandmodifiedaccordingtothecDNAse
4、quenceofP.xylostellaacetylcholineesterasegenesAChE1andAChE2.AllofthemwereformulatedandusedininsecticideactivityscreeningagainstP.xylostella.BioassaydatasuggestedthatSi-ace1_003andSi-ace2_001ata22concentrationof3mgcmdisplayedthebestinsecticidalactivitywith73.7%and89.0%,mortality,respec
5、tively.AdditionalbioassayswereusedtoobtaintheacutelethalconcentrationsofLC50andLC90forSi-ace2_001,whichwere53.66mg/mland759.71mg/ml,respectively.QuantitativeReal-timePCRwasusedtoconfirmsilencinganddetectedthatthetranscriptlevelsofP.xylostellaAChE2(PxAChE2)werereducedby5.7-foldcompared
6、tothecontrolgroup.Consequently,AChEactivitywasalsoreducedby1.7-fold.Finally,effectsofthesiRNAsontreatedplantsofBrassicaoleraceaandBrassicaalboglabrawereinvestigatedwithdifferentsiRNAdoses.OurresultsshowedthatSi-ace2_001hadnonegativeeffectsonplantmorphology,colorandgrowthofveinunderour
7、experimentalconditions.Conclusions:ThemostimportantfindingofthisstudyisthediscoverythatchemicallysynthesizedandmodifiedsiRNAcorrespondingtoP.xylostellaAChEgenescausesignificantmortalityoftheinsectbothunderlaboratoryandfieldconditions,whichprovidesanovelstrategytocontrolP.xylostellaand
8、todev
