4-Vinylanisole promotes conspecific interaction and acquisition of gregarious behavior in the migratory locust

Chemical signals from conspecifics are essential in insect group formation and maintenance. Migratory locusts use the aggregation pheromone 4-vinylanisole (4VA), specifically released by gregarious locusts, to attract and recruit conspecific individuals, leading to the formation of large-scale swarms. However, how 4VA contributes to the transition from solitary phase to gregarious phase remains unclear. We investigated the occurrence of locust behavioral phase changes in the presence and absence of 4VA perception. The findings indicated that solitary locusts require crowding for 48 and 72 h to adopt partial and analogous gregarious behavior. However, exposure to increased concentrations of 4VA enabled solitary locusts to display behavioral changes within 24 h of crowding. Crowded solitary locusts with RNAi knockdown of Or35, the specific olfactory receptor for 4VA, failed to exhibit gregarious behaviors. Conversely, the knockdown of Or35 in gregarious locusts resulted in the appearance of solitary behavior. Additionally, a multi-individual behavioral assay system was developed to evaluate the interactions among locust individuals, and four behavioral parameters representing the inclination and conduct of social interactions were positively correlated with the process of crowding. Our data indicated that exposure to 4VA accelerated the behavioral transition from solitary phase to gregarious phase by enhancing the propensity toward proximity and body contact among conspecific individuals. These results highlight the crucial roles of 4VA in the behavioral phase transition of locusts. Furthermore, this study offers valuable insights into the mechanisms of behavioral plasticity that promote the formation of locust swarms and suggests the potential for 4VA application in locust control.