Sex pheromone recognition and immunolocalization of three pheromone binding proteins in the black cutworm moth Agrotis ipsilon

Abstract

Insect pheromone binding proteins (PBPs) are believed to solubilize and transport hydrophobic sex pheromones across sensillum lymph to membrane-associated pheromone receptors. To address the molecular mechanisms of PBPs in insect pheromone perception, we undertook a systemic study on the PBPs of the black cutworm Agrotis ipsilon at transcript as well as protein level from tissue distribution and cellular localization to pheromone binding affinity. We cloned three full-length PBP genes AipsPBP1-3 from A. ipsilon antennae, and demonstrated that AipsPBP1–3 transcripts were highly expressed in male antennae. The electron microscopic examinations revealed at least six types of olfactory sensilla on male and female antenna: trichodea, chaetica, basiconica, coeloconica, squamiformia and Böhm bristles. The immunocytochemistry results demonstrated that AipsPBP1-3 proteins were strongly expressed in the sensillum lymph of the trichoid sensilla of male moth. The binding assays showed that AipsPBP1 had high binding affinities with the major sex pheromone components Z7-12:Ac and Z9-14:Ac among five related chemicals and was clustered together with the long trichoid sensilla-expressing LdisPBPs of Lymantria dispar. AipsPBP2 showed high binding affinities also with Z11-16:Ac. AipsPBP3 displayed a high affinity only with Z11-16:Ac. Our studies provide further detail evidences for the involvement of moth PBPs in pheromone discrimination and selective recognition of specific components of the female sex pheromone blends.