Molecular Characterization and Expression Pattern of Two Pheromone-Binding Proteins from Spodoptera litura (Fabricius)

Abstract

Pheromone perception is thought to be mediated by pheromone-binding proteins (PBPs) in the lymph surrounding the olfactory receptors. We cloned and characterized two PBP genes (SlitPBP1 and SlitPBP2) from the common cutworm, Spodoptera litura (F.; Lepidoptera: Noctuidae), which encode PBPs belonging to two different PBP groups. Western blot analysis of the crude antennal extracts with SexigPBP1 antibody revealed a single immunoreactive band (much stronger in male than in female) of approximately 16 kDa, in agreement with the calculated values for SlitPBPs. From genomic DNA, two introns and a similar exon/intron structural pattern were identified in each PBP genes, but the introns differed in length within and between PBP genes. The expression patterns of two SlitPBP genes, with respect to tissue distribution and sex, were further investigated by reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR. Although the two PBP genes were expressed only in the antennae of both sexes, reflecting the antennal specificity of PBPs, the transcription levels of PBP genes differed between the sexes and the genes. The transcription levels of SlitPBP1 and SlitPBP2 in females were only 2.1% and 7.0%, respectively, relative to those in males, and the levels of PBP2 compared with PBP1 were 31.4% and 95.3% in males and females, respectively. These differential expression levels might suggest different roles played by the two SlitPBPs in the perception of sex pheromone both in males and females.