Understanding the connection between sensory input and motor output is challenging. There is a need for good model systems to evaluate the molecular and neural mechanism of animal behavior. Here we show a new system for analyzing preference and plasticity of Drosophila egg-laying behavior. We found that selection of egg-laying site in flies is a good model to investigate the food preference in Drosophila. On sugar food with lower concentration, flies showed significant preference that was reduced upon the inhibition of olfactory pathway. Moreover, to examine the plasticity of egg-laying behavior, a new learning paradigm was devised. We associated light-dark cycle of egg-laying with different food conditions and found that flies could maintain egg-laying memory at least 3 days after training. This egg-laying system provides the basis for further study of molecular and neural mechanism underlying the relationship between environment and behavior.