The efficacy and high specificity of the RNA interference pathway has prompted its exploration as a potential molecular management tool for many insect pests, including the destructive southern pine beetle, Dendroctonus frontalis Zimmermann, in which gene knockdown and mortality via double-stranded RNAs (dsRNAs) have already been demonstrated in the laboratory. The nucleotide sequence of dsRNAs requires an exact match of at least 16 nucleotides with the targeted messenger RNA to trigger knockdown of that gene. This allows vital genes in a target pest to be silenced and mortality induced while reducing the probability of adverse effects in nontarget organisms. However, prior to utilization in forest ecosystems, demonstration of the specificity of dsRNAs through laboratory bioassays evaluating potential nontarget effects on model insects is required for proper risk assessment analyses. Consequently, we evaluated three SPB-specific dsRNAs for lethal effects, sublethal effects (larval growth rate, adult emergence or adult fecundity), and relative gene expression in three model nontarget insects representing key functional guilds, including a predator, herbivore, and pollinator. The SPB-specific dsRNAs had no effect on survival of our nontarget insects. Additionally, no sublethal effects were found and the gene expression analyses corroborated bioinformatic analyses in finding no gene knockdown. Our findings support the high specificity of RNAi technology and provide support for its development and deployment for protection of conifer forests against SPB with minimal nontarget concerns.

RNA 干扰途径的有效性和高度特异性促使其探索作为许多害虫的潜在分子管理工具，包括破坏性的南方松甲虫，Dendroctonus frontalisZimmermann，其中已经在实验室中证明了通过双链 RNA (dsRNA) 进行的基因敲低和死亡率。dsRNA 的核苷酸序列需要与目标信使 RNA 至少有 16 个核苷酸的精确匹配才能触发该基因的敲低。这使得目标害虫中的重要基因得以沉默并诱导死亡，同时降低对非目标生物产生不利影响的可能性。然而，在森林生态系统中利用之前，需要通过实验室生物测定评估对模式昆虫的潜在非目标影响来证明 dsRNA 的特异性，以进行适当的风险评估分析。因此，我们评估了三种 SPB 特异性 dsRNA 的致死效应、亚致死效应（幼虫生长率、成虫羽化或成虫繁殖力）、以及代表关键功能行会的三种模型非目标昆虫的相对基因表达，包括捕食者、食草动物和传粉者。SPB 特异性 dsRNA 对我们的非目标昆虫的存活没有影响。此外，没有发现亚致死效应，基因表达分析证实了生物信息学分析没有发现基因敲低。我们的研究结果支持 RNAi 技术的高度特异性，并为其开发和部署提供支持，以保护针叶林免受 SPB 的侵害，同时将非目标问题降至最低。没有发现亚致死效应，基因表达分析证实了生物信息学分析没有发现基因敲低。我们的研究结果支持 RNAi 技术的高度特异性，并为其开发和部署提供支持，以保护针叶林免受 SPB 的侵害，同时将非目标问题降至最低。没有发现亚致死效应，基因表达分析证实了生物信息学分析没有发现基因敲低。我们的研究结果支持 RNAi 技术的高度特异性，并为其开发和部署提供支持，以保护针叶林免受 SPB 的侵害，同时将非目标问题降至最低。