The lnc40545-miR-965-CYP307A1 regulation pathway mediates imidacloprid resistance in Rhopalosiphum padi (L.)

The competing endogenous RNA (ceRNA) hypothesis provides a novel perspective for investigating the post-transcriptional regulatory mechanisms of detoxification enzyme genes. In this study, a cytochrome P450 gene, Rhopalosiphum padi CYP307A1 was significantly upregulated in the imidacloprid-resistant strain (RP-R) compared to the imidacloprid-susceptible strain (RP-S), and silencing CYP307A1 significantly increased the susceptibility of R. padi to imidacloprid. Both CYP307A1 and the long non-coding RNA lnc40545 containing conserved microRNA response elements (MREs) for miR-965 were predicted by bioinformatic analysis, and was experimentally validated by using a dual-luciferase reporter assay, showing that miR-965 specifically targets and binds to both CYP307A1 and lnc40545. Furthermore, silencing lnc40545 resulted in the significant downregulation of CYP307A1 and a concomitant increase in imidacloprid sensitivity in R. padi. These results indicate that lnc40545 acts as a "molecular sponge" to sequester miR-965, thereby indirectly upregulating the expression of CYP307A1, suggesting the involvement of a lncRNA-mediated ceRNA mechanism in imidacloprid resistance in R. padi. It not only advances our understanding of insect detoxification metabolism but also provides a crucial theoretical basis for the integrated management of imidacloprid resistance.