Department of Biology, The University of Western Ontario, 1151 Richmond Street, London, ON, N6A 5B7, Canada.
Institut national de recherche pour l'agriculture, l'alimentation et l'environnement, 33882, Villenave d'Ornon, France.
Sci Rep. 2022 Aug 30;12(1):14791. doi: 10.1038/s41598-022-19231-3.
Environmental RNAi has been developed as a tool for reverse genetics studies and is an emerging pest control strategy. The ability of environmental RNAi to efficiently down-regulate the expression of endogenous gene targets assumes efficient uptake of dsRNA and its processing. In addition, its efficiency can be augmented by the systemic spread of RNAi signals. Environmental RNAi is now a well-established tool for the manipulation of gene expression in the chelicerate acari, including the two-spotted spider mite, Tetranychus urticae. Here, we focused on eight single and ubiquitously-expressed genes encoding proteins with essential cellular functions. Application of dsRNAs that specifically target these genes led to whole mite body phenotypes-dark or spotless. These phenotypes were associated with a significant reduction of target gene expression, ranging from 20 to 50%, when assessed at the whole mite level. Histological analysis of mites treated with orally-delivered dsRNAs was used to investigate the spatial range of the effectiveness of environmental RNAi. Although macroscopic changes led to two groups of body phenotypes, silencing of target genes was associated with the distinct cellular phenotypes. We show that regardless of the target gene tested, cells that displayed histological changes were those that are in direct contact with the dsRNA-containing gut lumen, suggesting that the greatest efficiency of the orally-delivered dsRNAs is localized to gut tissues in T. urticae.
环境 RNAi 已被开发为反向遗传学研究的工具,是一种新兴的害虫控制策略。环境 RNAi 能够有效下调内源性基因靶标的表达,假设其能够有效摄取 dsRNA 并对其进行加工。此外,通过 RNAi 信号的系统传播可以提高其效率。环境 RNAi 现已成为螯肢动物螨类(包括二斑叶螨)中基因表达操纵的成熟工具。在这里,我们集中研究了八个单一且普遍表达的基因,这些基因编码具有重要细胞功能的蛋白质。专门针对这些基因的 dsRNA 的应用导致整个螨虫体出现暗斑或无斑表型。这些表型与靶基因表达的显著减少有关,当在整个螨虫水平上评估时,其范围从 20%到 50%不等。用口服递送 dsRNA 处理螨虫的组织学分析用于研究环境 RNAi 的有效性的空间范围。尽管宏观变化导致两组体表型,但靶基因的沉默与明显的细胞表型有关。我们表明,无论测试的靶基因如何,显示组织学变化的细胞都是与含有 dsRNA 的肠道腔直接接触的细胞,这表明口服递送 dsRNA 的最大效率定位于 T. urticae 的肠道组织中。