CYP303A1 regulates molting and metamorphosis through 20E signaling in Nilaparvata lugens Stål (Hemiptera: Delphacidae)

Cytochrome P450s play a crucial role in the breakdown of external substances and perform important activities in the hormone system of insects. It has been understood that P450s were essential in the metabolism of ecdysteroids. CYP303A1 is a highly conserved CYP in most insects, but its specific physiological functions remain poorly understood in Nilaparvata lugens Stål. In this study, NlCYP303A1 was identified and highly expressed in the pre-molt stages, predominantly in the cuticle-producing tissues. Silencing of NlCYP303A1 caused a lethal phenotype with a molting defect. Moreover, the 20E titers, the expression levels of Halloween genes, and critical genes associated with the 20E signaling pathway in N. lugens nymphs were significantly decreased with the silencing NlCYP303A1. We further performed additional backfilling of 20E to rescue the RNAi effects on NlCYP303A1. The gene expression levels that were previously reduced caused by silencing NlCYP303A1 were significantly elevated. However, the molting defects of nymphs were not effectively improved. The results demonstrated NlCYP303A1 plays a crucial role in the molting and metamorphosis of N. lugens by regulating the 20E signaling pathway and cuticular formation, enhances the understanding of the functional role of CYP 2 clans, and identifies candidate gene for RNAi-based control of N. lugens.