Corazonin signaling modulates the synthetic activity of male accessory gland in Grapholita molesta

Although neuropeptide corazonin (Crz) has been identified in numerous insect species, the research about its function in regulation of reproduction is still in its infancy. Herein, we characterized the Crz (GmolCrz) and its receptor (GmolCrzR) to investigate their reproductive function in Grapholita molesta. Both molecular docking result and cell-based receptor activity assay showed that GmolCrz could interact with GmolCrzR. Additionally, spatial expression patterns of GmolCrz and GmolCrzR in males were evaluated. Knockdown of GmolCrz or GmolCrzR significantly lengthened copulation duration and decreased fertility in males. In these males, we found that the production of sperm was normal, while the content of accessory gland proteins (Acps) in the accessory gland (AG) was strongly diminished. Furthermore, knockdown of GmolCrz or GmolCrzR in males had no effect on sperm and Acps transfer to females. RNA-seq and gene expression analyses further confirmed that genes involved in serine-type endopeptidase activity were significantly downregulated in the AG upon GmolCrzR knockdown. Finally, sperm activation assays demonstrated that this process was disrupted in the spermatophore of females mated with GmolCrz or GmolCrzR knockdown males, which may cause the decreased fertility in males. Our findings provide new insights into the functions of Crz signaling in a Lepidopteran insect.