The olfactory recognition between leaf-cutter bee Megachile saussurei and alfalfa floral volatiles mediated by odorant binding protein 4 (MsauOBP4).

Megachile saussurei (Hymenoptera, Megachilidae) is a primary insect pollinator of alfalfa (Medicago sativa L.) in northwestern China. However, the mechanisms underlying the olfactory responses of M. saussurei induced by alfalfa volatiles is still unclear. Here, the interaction between MsauOBP4 and alfalfa floral volatiles was first elucidated. Results suggested that thirty-two alfalfa floral volatiles were identified and MsauOBP4 was successfully expressed with the consistent molecular mass as predicted results. MsauOBP4 displayed a broad binding spectrum to 32 volatiles, among which MsauOBP4 showed the strongest binding ability to (Z)-3-Hexen-1-ol. In the Y-tube olfactometer behavioral bioassay, M. saussurei elicited the most significant behavioral preference (Z)-3-Hexen-1-ol. MsauOBP4 showed an optimal binding feature to (Z)-3-Hexen-1-ol and valine was the key residue in binding the ligands. After silencing the MsauOBP4, the preference and EAG values of M. saussurei to (Z)-3-Hexen-1-ol were significantly decreased and selection rate of M. saussurei to alfalfa flowers dropped to 57.50 % from 83.33 %. These findings indicated that (Z)-3-Hexen-1-ol is a crucial component in the host location process mediated by MsauOBP4.