RNA Interference of Four Genes in Adult Bactrocera dorsalis by Feeding Their dsRNAs 英文参考文献.docVIP

RNAInterferenceofFourGenesinAdultBactrocera dorsalisbyFeedingTheirdsRNAs XiaoxueLi,MingyanZhang,HongyuZhang* State Key Laboratory of Agricultural Microbiology, Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control and Institute of Urban and HorticulturalPests,CollegeofPlantScienceandTechnology,HuazhongAgriculturalUniversity,Wuhan,Hubei,People’sRepublicofChina Abstract Background: RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner. Recentlysilencingthetargetgenethroughfeedinghasbeensuccessfullycarriedoutinmanyinsectspecies. Methodology/Principal Findings: Escherichia coli strain HT115 was genetically engineered to express dsRNA targeting genes that encode ribosomal protein Rpl19, V type ATPase D subunit, the fatty acid elongase Noa and a small GTPase Rab11.qRT-PCRshowedthatmRNAleveloffourtargetgeneswasreducedcomparedtods-egfpcontrolbyfeedingeither engineered bacteria or dsRNAs. The maximum down-regulation of each gene varied from 35% to 100%. Tissue specific examination indicated that RNAi could be observed not only in midgut but also in other tissues like the ovary, nervous systemandfatbody.Silencingofrab11throughingestionofdsRNAkilled20%ofadultflies.Eggproductionwasaffected through feeding ds-noa and ds-rab11 compared to ds-egfp group. Adult flies were continuously fed with dsRNA and bacteria expressing dsRNA for 14 days and up-regulations of target genes were observed during this process. The transcripts of noa showed up-regulation compared to ds-egfp control group in four tissues on day 7 after continuous feedingeitherdsRNAorengineeredbacteria.Themaximumover-expressionis21timescomparedtods-egfpcontrolgroup. Up-regulationofrab11mRNAlevelcouldbeobservedintestesonday7aftercontinuousbacteriatreatmentandinmidgut onday2afterds-rab11treatment.Thisphenomenoncouldalsobeobservedinrpl19groups. Conclusions:OurresultssuggestedthatitisfeasibletosilencegenesbyfeedingdsRNAandbacteriaexpressingdsRNAin