The neural representation of a sensory stimulus evolves with time, and animals keep that representation even after stimulus cessation (i.e., a stimulus "trace"). To contrast the memories of an odor and an odor trace, we here establish a rigorous trace conditioning paradigm in the fruit fly, Drosophila melanogaster. We modify the olfactory associative learning paradigm, in which the odor and electric shock are presented with a temporal overlap (delay conditioning). Given a few-second temporal gap between the presentations of the odor and the shock in trace conditioning, the odor trace must be kept until the arrival of electric shock to form associative memory. We found that memories after trace and delay conditioning have striking similarities: both reached the same asymptotic learning level, although at different rates, and both kinds of memory have similar decay kinetics and highly correlated generalization profiles across odors. In search of the physiological correlate of the odor trace, we used in vivo calcium imaging to characterize the odor-evoked activity of the olfactory receptor neurons in the antennal lobe. After the offset of odor presentation, the receptor neurons showed persistent, odor-specific response patterns that lasted for a few seconds and were fundamentally different from the response patterns during the stimulation. Weak correlation between the behavioral odor generalization profile in trace conditioning and the physiological odor similarity profiles in the antennal lobe suggest that the odor trace used for associative learning may be encoded downstream of the olfactory receptor neurons.