Israni B, Rajam M V
Department of Genetics, University of Delhi South Campus, New Delhi, India.
Insect Mol Biol. 2017 Apr;26(2):164-180. doi: 10.1111/imb.12277. Epub 2016 Nov 24.
RNA interference mediated gene silencing, which is triggered by double-stranded RNA (dsRNA), has become a important tool for functional genomics studies in various systems, including insects. Bacterially produced dsRNA employs the use of a bacterial strain lacking in RNaseIII activity and harbouring a vector with dual T7 promoter sites, which allow the production of intact dsRNA molecules. Here, we report an assessment of the functional relevance of the ecdysone receptor, insect intestinal mucin and sericotropin genes through silencing by dsRNA in two lepidopteran insect pests, Helicoverpa armigera and Plutella xylostella, both of which cause serious crop losses. Oral feeding of dsRNA led to significant reduction in transcripts of the target insect genes, which caused significant larval mortality with various moulting anomalies and an overall developmental delay. We also found a significant decrease in reproductive potential in female moths, with a drop in egg laying and compromised egg hatching from treated larvae as compared to controls. dsRNA was stable in the insect gut and was efficiently processed into small interfering RNAs (siRNAs), thus accounting for the phenotypes observed in the present work. The study revealed the importance of these genes in core insect processes, which are essential for insect development and survival.
由双链RNA(dsRNA)触发的RNA干扰介导的基因沉默,已成为包括昆虫在内的各种系统中功能基因组学研究的重要工具。细菌产生的dsRNA利用了一种缺乏RNaseIII活性且携带具有双T7启动子位点载体的细菌菌株,该载体允许产生完整的dsRNA分子。在此,我们报告了通过在两种鳞翅目害虫棉铃虫和小菜蛾中用dsRNA沉默来评估蜕皮激素受体、昆虫肠粘蛋白和促丝腺激素基因的功能相关性,这两种害虫都会导致严重的作物损失。口服dsRNA导致目标昆虫基因的转录本显著减少,这导致幼虫出现各种蜕皮异常并显著死亡,以及整体发育延迟。我们还发现雌性蛾的繁殖潜力显著下降,与对照相比,处理后的幼虫产卵量下降且卵孵化率受损。dsRNA在昆虫肠道中稳定,并被有效加工成小干扰RNA(siRNA),因此解释了本研究中观察到的表型。该研究揭示了这些基因在昆虫核心过程中的重要性,这些过程对昆虫的发育和生存至关重要。