AbstractMassive economic damage by spotted-wing drosophila (SWD), Drosophila suzukii, results from their unique egg laying behavior wherein a gravid fly pierces a ripening fruit to lay a number of eggs. Gravid SWD flies employ a complex suite of behaviors to find a fruit with the optimum firmness and chemistry. We investigated chemosensory cues potentially contributing to the oviposition behavior of SWD. In a series of experiments, we studied how the fruit ripeness and the underlying volatile chemistry influence oviposition. We tested the comparative attraction of three common fruits implicated in oviposition and determined raspberries to be most attractive in the trap choice assays that strictly measured olfactory preference. Since SWD oviposit in ripening fruits and appear to avoid the overripe fruit, we further evaluated the effect of ripeness on gravid fly attraction. Overripe fruits were significantly more attractive compared to the ripe fruits. The trap choice assays were repeated in an olfactory T-maze paradigm that provided a complex odor environment, potentially experienced by the gravid flies, and the results were mostly comparable. Since our behavioral paradigms indicated a clear olfactory preference for specific ripeness stages (ripe and overripe), we analyzed the constituent volatile odorants from the three ripening stages, revealing discrete odor profiles. Finally, we quantified the total soluble sugars and carbon dioxide concentrations from field-collected raspberries in underripe, ripe, and overripe conditions, revealing that the overripe stage is the most sugar-rich. Together, our results indicate unique chemosensory adaptations in gravid SWD flies for successfully exploiting optimal oviposition resources.