Acetyl-CoA, the precursor of sex pheromone biosynthesis in Helicoverpa armigera, is generated from glycolysis. As the first speed-limited enzyme in glycolysis, Hexokinase (HK) plays an important role in acetyl-CoA production. However, the function of HK in sex pheromone production remains unclear. This study employed H. armigera as material to explore the role of HK in sex pheromone production. Results demonstrated that the transcription profile of HaHK in female moth pheromone glands (PGs) was consistent with the release fluctuation of sex pheromone. Interference of HaHK prevented the increase of acetyl-CoA content induced by PBAN. Therefore, knockdown of HaHK in female PGs caused significant decreases in (Z)-11-hexadecenal (Z11-16:Ald) production, female capability to attract males, and mating rate. Furthermore, sugar feeding (5% sugar) increased the transcription and enzymatic activity of HK. Pheromone biosynthesis activating neuropeptide (PBAN) signal phospho-activated HaHK in PGs and Sf9 cells via protein kinase A (PKA), as shown by pharmacological inhibitor analysis. In general, our study confirmed that PBAN/cAMP/PKA signal activated HaHK, in turn promoted glycolysis to ensure the supply of acetyl-CoA, and finally facilitated sex pheromone biosynthesis and subsequent mating behavior.

The ladybird beetle, Henosepilachna vigintioctopunctata, is an oligophagous pest with significant economic impact. This pest causes considerable economic damage on numerous Solanaceae crops. Neuropeptides, along with their designated receptors, play a pivotal role in regulating diverse biological processes in insects, presenting a promising avenue for innovative pest management strategies. Herein, the transcriptome of the central nervous system (CNS) of H. vigintioctopunctata was sequenced. Overall, our analysis identified 58 neuropeptide precursor genes, from which 98 diverse mature peptides were predicted. Furthermore, 31 neuropeptide receptor genes belonging to three distinct classes were discovered, along with predictions for their potential ligands. Moreover, the expression patterns of these 58 neuropeptide genes across larval brain tissue, ventral nerve cord, and gut were evaluated using quantitative real-time PCR. Collectively, these findings will significantly contribute to future research focused on understanding the physiological functions and pharmacological characteristics of neuropeptides and their receptors in H. vigintioctopunctata. Ultimately, these insights may facilitate the development of targeted neuropeptide-based solutions for managing this pest affecting solanaceous plants.