The sensilla trichodea-biased EoblPBP1 binds sex pheromones and green leaf volatiles in Ectropis obliqua Prout, a geometrid moth pest that uses Type-II sex pheromones

Abstract

Pheromone-binding proteins (PBPs) are considered to play critical roles in sex pheromone detection. Lepidopteran moths can be divided into two taxa, those that use Type-I sex pheromones, such as C10-C18 unsaturated aldehydes, alcohols and acetates, and those that use Type-II pheromones, which are C17-C23 polyunsaturated hydrocarbons and their epoxide derivatives. To date, nearly all the characterized PBPs have been reported in moths with Type-I sex pheromones, and the physiological functions of PBPs in moths that use Type-II sex pheromones remains unclear. In the present study we functionally examine EoblPBP1 in Ectropis obliqua Prout, an important geometrid moth pest that uses Type-II sex pheromones. The phylogenetic analysis of the sequence indicated that EoblPBP1 clustered together with ScerPBP1, a geometrid PBP for detecting Type-II sex pheromones. Scanning electron microscopy showed that E. obliqua moths of both sexes mainly had six types of antennal sensilla, including two types of sensilla trichodea, Str-I and Str-II, sensilla basiconica (Sba), sensilla styloconica (Sst), sensilla chaetica (Sch) and sensilla auricillica (Sau). Of these, Str-I was confirmed to be male moth-specific and had five different subtypes. Fluorescence in situ hybridization revealed that EoblPBP1 was primarily expressed at the base of Str-I. A comparative binding assay showed that recombinant EoblPBP1 bound three sex pheromone components of E. obliqua, demonstrating its involvement in the detection of Type-II sex pheromones. Besides, EoblPBP1 also highly bound unsaturated acetates pheromones and the green leaf volatiles. These results indicate that PBP1 is associated with detecting Type-II sex pheromones in E. obliqua but cannot differentiate Type-II sex pheromones from Type-I sex pheromones or green leaf volatiles. Our findings provide a foundation for further study on molecular basis of Type-II sex pheromone recognition in lepidopteran moths.

Keywords: Fluorescence in situ hybridization; Fluorescence-based competitive binding assay; Pheromone-binding protein; Sensilla types; Type-II sex pheromones.