Identification and functional analysis of gut dsRNases in the beet armyworm Spodoptera exigua

Abstract

RNA interference (RNAi)-based products have the potential to significantly contribute to insect pest control. However, RNAi efficiency varies widely among different insect orders, particularly in Lepidoptera, where it is often low. One key factor affecting RNAi efficiency is the presence of double-stranded ribonuclease (dsRNase) in the digestive tract, which can degrade dsRNA prior to uptake by gut cells. In this study, four dsRNases were identified in the beet armyworm, Spdoptera exigua, of which two were highly expressed gut dsRNases, SedsRNase1 and SedsRNase2. To assess their effect on dsRNA degradation activity via the oral route, CRISPR/Cas9-based gene editing was employed to knock out these gut dsRNases. The results indicate that all mutant strains, including SeKO1 (knockout SedsRNase1), SeKO2 (knockout SedsRNase2), and SeKO1KO2 (knockout SedsRNase1 and SedsRNase2), showed significantly decreased dsRNA degradation activity, particularly in the SeKO1KO2 mutant strain, where the weakest degradation occurred in both the gut and whole body. Additionally, we noticed that the lack of gut SedsRNases led to a slight extended developmental period and reduced reproductive capacity in S. exigua. Collectively, these findings deepen our understanding of gut SedsRNases and how they can impact the biology of the beet armyworm and can support the exploration dsRNA-based approaches for pest control.