The Chemoreceptive Molecular Mechanism Underlying CSP-Mediated Recognition of Seed Elaiosome from Stemona tuberosa by...

As crucial natural predators, hornets contribute to ecosystem function by preying on agricultural and forest pests and facilitating plant pollination. However, the predatory preference of hornets for honeybees poses a significant threat to honeybee pollination and the development of the beekeeping industry. Foraging and pollination behaviors in hornets are largely governed by a sensitive olfactory system, but their olfactory molecular mechanisms remain poorly understood.

VvelCSP1 and VvelCSP4 were successfully expressed in the prokaryotic expression system and purified by Ni-NTA affinity chromatography column. Fluorescence competitive binding assays were employed to evaluate their binding affinities to volatile compounds derived from the seed elaiosome of Stemona tuberosa and honeybees. Molecular docking was further performed to analyze key residues and interaction patterns within the binding pockets.

Fluorescence competitive binding assays showed that both proteins prefer long-chain alkanes yet exhibit significant substrate selectivity and high ligand specificity. VvelCSP1 specifically binds to hexacosane, while VvelCSP4 specifically recognizes docosane. Molecular docking results demonstrated that the binding process between VvelCSP1, VvelCSP4 and their respective ligands is dominated by hydrophobic interactions.

This study provides functional evidence for investigating the olfactory molecular regulation mechanisms underlying hornet-mediated seed dispersal. These findings establish a foundation for potential applications of hornets in plant propagation, biological pest control, crop pollination and ecological balance maintenance in agroforestry systems.