Leptinotarsa cap ‘n’ collar isoform C/Kelch-like ECH associated protein 1 signaling is critical for the regulation of ecdysteroidogenesis in the larvae

Drosophila cap 'n' collar isoform C (CncC) and Kelch-like ECH associated protein 1 (Keap1) regulate metamorphosis by transcriptional control of a subset of genes involved in ecdysteroidogenesis, 20-hydroxyecdysone (20E) signaling, and juvenile hormone (JH) degradation. In the present paper, we found that prothoracicotropic hormone signal was required for the activation of LdCncC and LdKeap1 in Leptinotarsa decemlineata. Moreover, RNA interference of LdCncC or LdKeap1 in the fourth-instar larvae delayed development. As a result, the treated larvae obtained heavier larval and pupal fresh weights and had larger body sizes than the controls. Furthermore, knockdown of LdCncC or LdKeap1 significantly reduced the mRNA levels of four ecdysone biosynthetic genes (Ldspo, Ldphm, Lddib and Ldsad), lowered 20E titer and decreased the transcript levels of five 20E response genes (LdEcR, LdUSP, LdE75, LdHR3 and LdFTZ-F1). However, the expression of two JH epoxide hydrolase genes and JH contents were not affected in the LdCncC and LdKeap1 RNAi larvae. Dietary supplementation with 20E shortened the developmental period to normal length, rescued the larval and pupal body mass rises, and recovered or even overcompensated the expression levels of the five 20E response genes in either LdCncC or LdKeap1 RNAi hypomorphs. Therefore, LdCncC/LdKeap1 signaling regulates several ecdysteroidogenesis genes, and consequently 20E pulse, to modulate the onset of metamorphosis in L. decemlineata.