Targeting Co-Safe RNAi Genes: Identification of Chitin Synthase and β-1,3-Glucanase for Sustainable Control of the Invasive Pest <i>Tuta absoluta</i>

RNA interference (RNAi) has emerged as a promising method for pest control. While RNA interference shows promise for pest control, its environmental risk assessment remains essential. Tuta absoluta is a significant invasive pest posing a serious threat to the tomato industry in China, and the study is targeted at the pest. We evaluated two chitin synthase genes ( C hs1, Chs2 ) and a β-1,3-glucanase gene ( Beita ) as RNAi targets in T. absoluta, with particular emphasis on ecological safety. DsRNA exposure induced significant larval mortality (53–69%), phenotypic abnormalities, and midgut disruption. Most importantly, comprehensive environmental safety assessment confirmed no homologous genes in humans or key nontarget organisms ( Nesidiocoris tenuis, Harmonia axyridis, Trichogramma exiguum, Episyrphus balteatus ), and critically, no adverse effects on their survival, pollination, predation, or parasitism capacities. This study identifies three effective RNAi targets and provides substantial evidence for the environmental safety of these dsRNAs, supporting their sustainable application in T. absoluta management.