Silencing of Aphid Genes by dsRNA Feeding from Plants 英文参考文献.docVIP

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Silencing of Aphid Genes by dsRNA Feeding from Plants 英文参考文献.doc

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Silencing of Aphid Genes by dsRNA Feeding from Plants 英文参考文献

SilencingofAphidGenesbydsRNAFeedingfromPlants MarcoPitino1,2.,AlexanderD.Coleman1.,MassimoE.Maffei2,ChristopherJ.Ridout1,SaskiaA. Hogenhout1* 1Department of Disease and Stress Biology, The John Innes Centre, Norwich Research Park, Norwich,United Kingdom, 2Plant Physiology Unit, Department of Plant Biology,InnovationCentre,UniversityofTurin,Turin,Italy Abstract Background: RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphonpisum)genescanbeachievedthroughdirectinjectionofdouble-strandedRNA(dsRNA)orsmall-interfering RNAs(siRNA)intothepeaaphidhemolymphorbyfeedingtheseinsectsonartificialdietscontainingthesmallRNAs. Methodology/PrincipalFindings:Inthisstudy,wehavedevelopedtheplant-mediatedRNAitechnologyforaphidstoallow forgenesilencingintheaphidnaturalenvironmentandminimizehandlingoftheseinsectsduringexperiments.Thegreen peach aphid M. persicae was selected because it has abroad plant host range that includes the model plants Nicotiana benthamianaandArabidopsisthalianaforwhichtransgenicmaterialscanrelativelyquicklybegenerated.WetargetedM. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA correspondingtothesegenesandonA.thalianaplantsstablyproducingthedsRNAs.MpC002andRack-1expressionwere knockeddownbyupto60%ontransgenicN.benthamianaandA.thaliana.Moreover,silencedM.persicaeproducedless progenyconsistentwiththesegeneshavingessentialfunctions. Conclusions/Significance: Similar levels of gene silencing were achieved in our plant-mediated RNAi approach and publishedsilencingmethodsforaphids.Furthermore,theN.benthamianaleafdiskassaycanbedevelopedintoascreento assess which genes are essential for aphid survival on plants. Our re