简介:EliminationoftheCRISPR/Cas9constructsineditedplantsisaprerequisiteforassessinggeneticstability,conductingphenotypiccharacterization,andapplyingforcommercializationoftheplants.However,removaloftheCRISPR/Cas9transgenesbygeneticsegregationandbybackcrossislaboriousandtimeconsuming.WepreviouslyreportedthedevelopmentofthetransgenekillerCRISPR(TKC)technologythatusesapairofsuicidegenestotriggerself-eliminationofthetransgeneswithoutcompromisinggeneeditingefficiency.TheTKCtechnologyenablesisolationoftransgene-freeCRISPR-editedplantswithinasinglegeneration,greatlyacceleratingcropimprovements.Here,wepresentedtwonewTKCvectorsthatshowgreatefficiencyinbotheditingthetargetgeneandinundergoingself-eliminationofthetransgenes.ThenewvectorsreplacedtheCaMV35SpromoterusedinourpreviousTKCvectorwithtworicepromoterstodriveoneofthesuicidegenes,providingadvantagesoverourpreviousTKCvectorundercertainconditions.Thevectorsreportedhereofferedmoreoptionsandflexibilitytoconductgeneeditingexperimentsinrice.
简介:Apopulationofricebrownplanthopper(BPH)NilaparvatalugenscollectedfromapaddyfieldinHangzhouwassuccessivelyrearedonsusceptiblericeTaichungNative1(TN1)inalaboratoryfreefrominsecticidesformorethan14generations.Thechangesinsusceptibilitytoinsecticidesandecologicalfitnessondifferentresistantricevarietiesweremonitoredineachgeneration.Theresistanceratiotoimidaclopridsharplydeclinedwiththesuccessionofrearinggenerationswithoutinsecticidesfrom359.94-foldatF1to6.50-foldatF14comparedwiththesusceptiblestrain,andtheresistanceratiotochlorpyrifoswasfrom9.90-foldatF1to5.94-foldatF14.Nymphaldurationandweightsofnewlyhatchedfemaleadultsweresignificantlyaffectedbyricevariety,generationandtheirinteractions,butnymphalsurvivalwassignificantlyaffectedbythegenerationonly.Theratioofbrachypterousadultsinmaleswasaffectedbythegenerationandgeneration×varietyinteraction,whereasnodifferencewasfoundinfemales.Nymphaldurationextendedwithincreasinggenerations,andthefemalenymphaldurationwasshorterinthesusceptiblevarietyTN1thanthoseintheresistantvarietiesIR26andIR36.Inaddition,thefemaleadultweightinTN1washigherthanthoseinIR26andIR36.TheseresultsindicatedthattheresistanceoffieldBPHpopulationtoinsecticideswasreversedafterseveralgenerationsofno-exposuretoinsecticides,andtheecologicalfitnessinTN1washigherthanthoseinIR26andIR36.ThesefindingssuggestedtherationalandreduceduseofinsecticidesincombinationwiththemanipulationofresistantricevarietieswouldbeeffectiveforBPHmanagement.