Pheromones emitted by both female and male moths regulate coordination between the sexes for Agriphila aeneociliella (Lepidoptera: Crambidae).

The complex and efficient sex pheromone communication system in insects is essential for reproduction and species reproductive isolation. In moths, sex pheromone communication starts with male attraction to compounds emitted by females; only a few species act in the reverse. However, how the pheromones that are emitted by both sexes co-regulate and coordinate mate finding and mating remains unknown. Here, we identified both the male and female pheromones of Agriphila aeneociliella (Lepidoptera: Crambidae) and demonstrated their efficiency in manipulating behavioral responses of the opposite sex. Combining data from analysis of gas chromatography-electroantennogram detection (GC-EAD), gas chromatography-mass spectrometry (GC-MS), and olfactory behavior assays, the female pheromone of A. aeneociliella was identified as (Z,Z,Z)-9,12,15-octadecatrienal and (Z)-9-hexadecenyl acetate, while the male pheromone was determined to be 1-nonanal. Both of the two individual components of the female pheromone and their binary mixture were significantly attractive to males, and the 1-nonanal male pheromone induced strong electrophysiological responses in females and induced attraction of females in a Y-tube olfactory test. Depending on the concentration of 1-nonanal, its addition to the binary mixture of the female pheromone either enhanced (10 -3 , 10 -2 μg/μL) or reduced (1 μg/μL) the aphrodisiac effect of the mixture on males. In wind tunnel bioassays, different concentrations of pheromones, including the binary mixture of female pheromone and the mixture of male and female pheromones, had significant effects on male behavior. Our findings suggested that the blend of both female and male pheromones plays a significant role in sexual communication system in some moths. (Z,Z,Z)-9,12,15-octadecatrienal and (Z)-9-hexadecenyl acetate are female pheromones of A. aeneociliella. Male pheromone 1-nonanal not only manipulates female acceptance but affects male mating behavior through acting together with female pheromones. This article is protected by copyright. All rights reserved.