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RNAi-mediated mortality of the whitefly through transgenic expression of double-stranded RNA homologous to acetylcholinesterase and ecdysone receptor in tobacco plants

机译:通过与烟草中乙酰胆碱酯酶和蜕皮激素受体同源的双链RNA转基因表达,RNAi介导的粉虱死亡

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摘要

The whitefly Bemisia tabaci (Genn.) is a pest and vector of plant viruses to crop and ornamental plants worldwide. Using RNA interference (RNAi) to down regulate whitefly genes by expressing their homologous double stranded RNAs in plants has great potential for management of whiteflies to reduce plant virus disease spread. Using a Tobacco rattle virus-derived plasmid for in planta transient expression of double stranded RNA (dsRNA) homologous to the acetylcholinesterase (AChE) and ecdysone receptor (EcR) genes of B. tabaci, resulted in significant adult whitefly mortality. Nicotiana tabacum L. plants expressing dsRNA homologous to B. tabaci AChE and EcR were constructed by fusing sequences derived from both genes. Mortality of adult whiteflies exposed to dsRNA by feeding on N. tabacum plants, compared to non-dsRNA expressing plants, recorded at 24-hr intervals post-ingestion for three days, was > 90% and 10%, respectively. Analysis of gene expression by real time quantitative PCR indicated that whitefly mortality was attributable to the down-regulation of both target genes by RNAi. Results indicated that knock down of whitefly genes involved in neuronal transmission and transcriptional activation of developmental genes, has potential as a bio-pesticide to reduce whitefly population size and thereby decrease virus spread.
机译:烟粉虱烟粉虱(Bemisia tabaci,Genn。)是一种植物病毒的害虫和媒介,对全世界的农作物和观赏植物都是如此。使用RNA干扰(RNAi)通过在植物中表达它们的同源双链RNA来下调粉虱基因具有巨大的潜力来管理粉虱以减少植物病毒疾病的传播。使用烟草拨浪鼓病毒衍生的质粒在植物中瞬时表达与烟粉虱的乙酰胆碱酯酶(AChE)和蜕皮激素受体(EcR)基因同源的双链RNA(dsRNA),导致成年粉虱死亡。通过融合衍生自两个基因的序列,构建表达与烟粉虱AChE和EcR同源的dsRNA的烟草。与不表达dsRNA的植物相比,通过摄取烟草植物而暴露于dsRNA的成年粉虱的死亡率,分别在摄取后24小时间隔三天,分别> 90%和10%。通过实时定量PCR对基因表达的分析表明,粉虱的死亡率可归因于RNAi对两个靶基因的下调。结果表明,敲除参与神经元传递和发育基因转录激活的粉虱基因,具有作为生物农药的潜力,可以减少粉虱种群的数量,从而减少病毒的传播。

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