Molecular and functional characterization of three odorant-binding proteins from the wheat blossom midge, Sitodiplosis mosellana.

Abstract

Sitodiplosis mosellana, a periodic but devastating wheat pest, relies on wheat spike volatiles as a cue in selecting hosts for oviposition. Insect odorant-binding proteins (OBPs) are thought to play essential roles in filtering, binding and transporting hydrophobic odorant molecules to specific receptors. To date, the molecular mechanisms underlying S. mosellana olfaction are poorly understood. Here, three S. mosellana antenna-specific OBP genes, SmosOBP11, 16 and 21, were cloned and bacterially expressed. Binding properties of the recombinant proteins to 28 volatiles emitted from wheat spikes were investigated using fluorescence competitive binding assays. Sequence analysis suggested that these SmosOBPs belong to the Classic OBP subfamily. Ligand-binding analysis showed that all three SmosOBPs preferentially bound alcohol, ester and ketone compounds, and SmosOBP11 and 16 also selectively bound terpenoid compounds. In particular, the three SmosOBPs had high binding affinities (Ki < 20 μmol/L) to 3-hexanol and cis-3-hexenylacetate that elicited strong electroantennogram (EAG) response from female antennae. In addition, SmosOBP11 displayed significantly higher binding (Ki < 8 μmol/L) than SmosOBP16 and 21 to 1-octen-3-ol, D-panthenol, α-pinene and heptyl acetate which elicited significant EAG response, suggesting that SmosOBP11 plays a major role in recognition and transportation of these volatiles. These findings have provided important insight into the molecular mechanism by which S. mosellana specifically recognizes plant volatiles for host selection, and have facilitated identification of effective volatile attractants that are potentially useful for pest monitoring and trapping.