The limited regulatory roles of juvenile hormone degradation pathways in reproductive diapause preparation of the cabbage beetle, Colaphellus bowringi.

The absence of juvenile hormone (JH) induces reproductive diapause in many insects. Downregulated JH biosynthesis is considered the critical factor that decreases the JH level. However, whether upregulated JH degradation could also contribute to JH deficiency and then induce reproductive diapause remains poorly known. In this work, we used the cabbage beetle, Colaphellus bowringi, as the model and investigated the function of JH degradation genes in reproductive diapause preparation via RNAi-mediated loss-of-function. Three JH degradation genes, including JH epoxide hydrolase 1 (JHEH1), JHEH2, and JH diol kinase (JHDK), were cloned, and the deduced proteins contained the conserved domains and motifs well-identified in the same proteins of other insects. JHEH1, JHEH2, and JHDK were all highly expressed in the diapause-destined (DD) females compared to the non-diapause-destined (NDD) females. Knocking down these JH degradation genes in DD females elevated JH signaling, indicated by increased expressions of JH-inducible genes and vitellogenin genes, but did not induce ovary development. Meanwhile, the knockdown of JH degradation genes significantly suppressed diapause-associated phenotypic traits, including fat storage, and the expression of genes related to fat storage and stress tolerance. A photoperiodic transfer experiment suggested that high expression of JHEH2 in DD female was preprogrammed by long day during the larval stage. Knockdown of JH signaling through Methoprene-tolerant and Krüppel homolog 1 induced high expression of JHEH2 in DD females. Considering that RNAi of JH degradation genes affected some diapause-related traits in DD females but failed to induce the non-diapause phenotype in full, we conclude that JH degradation pathways play limited regulatory roles in the reproductive diapause preparation of C. bowringi.