Discrimination of oilseed rape volatiles by the honeybee: combined chemical and biological approaches

Honeybees (Apis mellifera L.) were individually subjected to a classical conditioning procedure in order to obtain an olfactory conditioned proboscis extension response. To relate the behavioural response directly to antennal detection abilities, a technique was developped for coupling proboscis extension responses and electroantennogram recordings, with the stimulation being provided by the effluent of a gas chromatograph (GC). Bees were conditioned with a six-component mixture being part of oilseed rape (Brassica napus L.) floral volatiles, and tested with the individual components separated by GC. Responses of the conditioned bees were compared to those of unconditioned bees. No behavioural response was obtained in the control group, neither to the individual components nor to the mixture. Conditioning induced behavioural responses for three components, and an increase of electroantennogram responses for all components. A second experiment was conducted with an air entrainment extract of oilseed rape flower volatiles. Behavioural responses of conditioned and unconditioned bees were recorded. Responses obtained from conditioned bees tested with the air entrainment extract showed six groups of behaviourally active GC peaks. Unconditioned bees showed the same pattern of responses but at a lower level. The coupled technique described here appears to be a reliable tool for locating active components in a synthetic as well as in a natural mixture of floral volatiles. The effects of conditioning on odour discrimination and on its sensory correlates are discussed.