Insect olfaction system plays a key role in the foraging food, pollination, mating, oviposition, reproduction and other insect physiological behavior. Odorant binding protein are widely found in the various olfactory sensilla of different insect antennae and involved in chemical signals discrimination from natural environment. In this study, a novel OBP gene,MvitOBP3 is identified from the legume pod borer, Maruca vitrata, which it mainly harms important legume vegetables including cowpea, soybean and lablab bean. Real-time PCR results demonstrated that MvitOBP3 gene was abundantly expressed in the antennal tissue of M. vitrata, while low levels were distributed in the head, thorax, abdomen, leg and wing of adult moths. The recombinant OBP3 protein was purified using the prokaryotic expression and affinity chromatography system. Fluorescence competitive binding experiments indicated that that MvitOBP3 protein exhibited greater binding affinities with host-plant flower volatiles including Butanoic acid butyl ester, Limonene, 1H-indol-4-ol and 2-methyl-3-phenylpropanal, highlighting they may have attractant activities for the oviposition of female moths on the legume vegetables.
Fig: Evolutionary analysis of MvitOBP3 protein with other Lepidopteran insect OBPs. Genbank accession numbers: LsinOBP (VVC93553.1), PmacOBP (CAF4841106.1), BmorOBP (NP001040212.1), CsasOBP8 (AYD42183.1), LbotOBP32 (AXF48729.1), PxutOBP (NP001299556.1), DpleOBP58a-like(XP 032522559.1), GmelOBP56a (XP026763748.1), PpelOBP (WP143520259.1), KpneOBP (WP142379627.1), EoblOBP16 (ALS03864.1), DpunOBP46 (ARO70205.1), DpunOBP22 (ARO70181.1), DhouOBP (AII00983.1), DkikOBP (AII01006.1), CsupOBP2 (AGK24578.1), HvitOBP6 (AZB49387.1), CpinOBP20 (QEE82719.1), CpunOBP(AMY16434.1), CmedOBP20 (ALT31650.1), HrhoOBP (QGN01742.1), PxylOBP32 (ANG08535.1), CsinOBP18 (QGN03648.1), EhipOBP (AOG12864.1), VtamOBP (XP026497454.1), AplaOBP (CAB3233796.1), SexiOBP9 (AGH70105.1), SlitOBP2 (XP 022815401.1), AdisOBP (QCF41928.1), MsepOBP16 (AWT22226.1), HvirOBP (PCG75714.1),HarmOBP17 (AFI57166.1), HassOBP17 (AGC92792.1), OfurOBP (XP028157948.1), SfruOBP (KAF9813770.1).
Moreover, protein homology modeling and molecular docking analysis revealed that there are six amino acid sites of MvitOBP3 involved in the binding of the host-plant volatiles. These findings will further promote to understand the key role of odorant binding protein during host perception and oviposition of M. vitrata moths, which improve the efficiency of semiochemical-based prevention and monitoring for this pest in the legume vegetables field.
Ai, H., Liu, Y., Long, G. et al. Functional characteristics of a novel odorant binding protein in the legume pod borer, Maruca vitrata. Sci Rep 11, 14027 (2021). https://doi.org/10.1038/s41598-021-93382-7