Exploring the functional profiles of odorant binding proteins crucial for sensing key odorants in the new leaves of coconut palms in Rhynchophorus ferrugineus

The red palm weevil (RPW), Rhynchophorus ferrugineus (Curculionidae: Coleoptera) is a highly destructive global pest of coconut trees, with a preference for laying its eggs on new leaves. Females can identify where to lay eggs by using their sense of smell to detect specific odorants found in new leaves. In this study, we focused on the two odorants commonly found in new leaves by GC-MS: trans, trans-2,4-nonadienal and trans-2-nonenal. Our behavioral assays demonstrated a significant attraction of females to both of these odorants, with their electrophysiological responses being dose-dependent. Furthermore, we examined the expression patterns induced by these odorants in eleven RferOBP genes. Among them, RferOBP3 and RferOBP1768 exhibited the most significant and simultaneous upregulation. To further understand the role of these two genes, we conducted experiments with females injected with OBP-dsRNA. This resulted in a significant decrease in the expression of RferOBP3 and RferOBP1768, as well as impaired the perception of the two odorants. A fluorescence competitive binding assay also showed that both RferOBPs strongly bound to the odorants. Additionally, sequence analysis revealed that these two RferOBPs belong to the Minus-C family and possess four conserved cysteines. Molecular docking simulations showed strong interactions between these two RferOBPs and the odorant molecules. Overall, our findings highlight the crucial role of RferOBP3 and RferOBP1768 in the olfactory perception of the key odorants in coconut palm new leaves. This knowledge significantly improves our understanding of how RPW females locate sites for oviposition and lays the foundation for future research on the development of environmentally friendly pest attractants.