miR-9a and miR-10482-5p regulate the expression of chitin synthase and chitinase genes, enhancing lufenuron tolerance in Spodoptera frugiperda.

Spodoptera frugiperda is a significant agricultural pest, severely impacting the yield and quality of grain. Chitin is the momentous component of exoskeletons, which has a significant impact on the growth and development of insects. Our previous study found that exposure to lufenuron can reduce the expression of chitinase gene (SfCHT5) and increase the expression of chitin synthase gene (SfCHSB), two key genes for chitin synthesis in S. frugiperda. However, the post-transcriptional regulatory mechanisms of these key genes in S. frugiperda remain unclear. With miRNA as the entry point, target site prediction, dual luciferase reporter assays, and mimics/inhibitors injection were used to explore the post-transcriptional regulatory mechanism of SfCHSB and SfCHT5. The combined results confirm that miR-10482-5p targets SfCHT5 and regulates its expression, while miR-9a targets and regulates the expression of both SfCHT5 and SfCHSB. Additionally, injection of miR-10482-5p and miR-9a mimic significantly reduced the mortality rates of S. frugiperda treated with lufenuron, increased pupation rate, and emergence rate. These findings underscore the critical regulatory role of miR-10482-5p and miR-9a in modulating the expression of key genes involved in chitin synthesis, thereby enhancing the tolerance of S. frugiperda to lufenuron and influencing its growth and development. In summary, this study preliminarily elucidates the post-transcriptional regulatory mechanisms of miRNA-mediated SfCHSB and SfCHT5 expression.