Drosophila have evolved strong mutualistic associations with yeast communities that best support their growth and survival, resulting in the development of novel niches. It has been suggested that flies recognize their cognate yeasts primarily based on the rich repertoire of volatile organic compounds (VOCs) derived from the yeasts. Thus, it remained an exciting avenue to study whether fly spp. detect and discriminate yeast strains based on odor alone, and if so, how such resolution is achieved by the olfactory system in flies. We used two fly species known to exploit different niches and harboring different yeasts, D. suzukii (a pest of fresh fruit) and D. melanogaster (a saprophytic fly and a neurogenetic model organism). We initially established the behavioral preference of both fly species to six Drosophila-associated yeasts; then chemically analyzed the VOC profile of each yeast which revealed quantitative and qualitative differences; and finally isolated and identified the physiologically active constituents from yeast VOCs for each drosophilid that potentially define attraction. By employing chemical, behavioral, and electrophysiological analyses, we provide a comprehensive portrait of the olfactory neuroethological correlates underlying fly-yeast coadaptation in two drosophilids with distinct habitats.