Supplementary material from "Fruit flies can learn non-elemental olfactory discriminations"

Associative learning allows animals to establish links between stimuli based on their concomitance. In the case of Pavlovian conditioning, a single stimulus A (the conditional stimulus, CS) is reinforced unambiguously with an unconditional stimulus (US) eliciting an innate response. This conditioning constitutes an ‘elemental’ association enabling to elicit a learnt response from A⁺ without US presentation after learning. However, associative learning may involve a ‘complex’ CS composed of several components. In that case, the compound may predict a different outcome than the components taken separately, leading to an ambiguity and requiring the animal to perform a so-called non-elemental discrimination. Here, we focus on such a non-elemental task, the negative patterning (NP) problem, and provide the first evidence of NP solving in Drosophila. We show that Drosophila learn to discriminate a simple component (A or B) associated with electric shocks (+) from an odour mixture composed either partly (called ‘feature-negative discrimination’ A⁺ versus AB−) or entirely (called ‘NP’ A⁺B⁺ versus AB⁻) of the shock-associated components. Furthermore, we show that conditioning repetition results in a transition from an elemental to a configural representation of the mixture required to solve the NP task, highlighting the cognitive flexibility of Drosophila.