Stored perfume dynamics and consequences for signal development in male orchid bees.

Male orchid bees (Euglossini) collect volatiles from their environment to concoct species-specific "perfumes", which are later emitted at mating sites. Intensity, complexity or composition of perfumes may encode age (survival) of a male, but how the individual perfume develops over time needs to be clarified. We investigated chemical changes during storage in leg pockets. We injected a mixture of eight perfume compounds into pockets of Euglossa imperialis and only the two most volatile compounds decreased over 12 days. Using a different approach we found significant shifts in quantities of naturally occurring perfume compounds of Euglossa championi over 10 days, with the strongest decreases (up to 70% peak area) in highly volatile minor compounds, e.g. monoterpenes, and noteworthy increases (up to 40%) in some sesquiterpenoids. Corresponding shifts were observed in legs of dried bees, suggesting that no metabolic activity is required for the observed changes to occur. Our results confirm that male orchid bees are generally good at preserving collected perfumes. However, subtle shifts towards heavier compounds in blends may occur over the lifetime of individual bees, e.g. due to evaporation or in-pocket chemical reaction, with old males acquiring a more pronounced base note in their seasoned perfumes.

Acquisition of species-specific perfume blends: influence of habitat-dependent compound availability on odour choices of male orchid bees (Euglossa spp.).

Male orchid bees (Euglossini, Apidae, Hymenoptera) expose species-specific blends of volatile chemicals (perfume bouquets) during their courtship display. The perfumes are acquired by collecting fragrant substances from environmental sources, which are then accumulated in specialised hind leg pouches. To balance the perfume composition, the males need to find and collect the required substances in specific relative amounts while facing seasonal, local or habitat-dependent differences in compound availability. Experience-dependent choice of odours, i.e. 'learned avoidance' of recently collected components, has been proposed as the mechanism that mediates the accumulation of the stereotypical compound ratios. In the present study, we used the presence of certain compounds in male hind leg pouches as proxy for the respective local compound availability, and investigated whether differences in content are correlated with differences in chemical choice assays. Our results suggest that volatile availability differs between localities (n = 16) as well as habitats (n = 2; coastal vs. inland) across the Yucatán peninsula, Mexico, for both studied species. Male Euglossa dilemma showed a pronounced preference for benzyl benzoate and eugenol at locations where those compounds were rare in hind leg extracts, as predicted by the learned avoidance model. No equivalent correlations were found for Euglossa viridissima. This is the first study to combine chemical analyses of perfumes with bioassays of odour choice. It strengthens the view that negative feedback from collected odours modifies future chemical choice and helps males to acquire specific perfume blends.