Global warming exacerbates rainfall irregularity and causes a decline in water resources. Drought is one of the main limiting parameters in agricultural and fruit production. In this study, we established a standard method for measuring drought tolerance in Fragaria , the simplest genomic model in the Rosaceae, a plant family comprising important fruit species like almonds, apples, plums, etc. A quick screening method for monitoring Fragaria genotypes in response to dehydration involved conducting a short-term water deficit experiment. We measured two eco-physiological parameters associated with leaf water status, leaf relative water content (RWC), and leaf water loss rate (WLR) in a total of 20 strawberry cultivars (Fragaria × ananassa Duch.) from different breeding programs. The plants included two ecotypes of the European diploid species, F. vesca, and one American octaploid species, F. chiloensis. Fragaria genotypes responded to drought stress, as measured by WLR and RWC, and DNA fingerprints further described select Fragaria genotypes using AFLP and candidate gene EST markers. We revealed correlations among specific DNA markers, leaf WLR, and RWC while navigating the possibility of using association mapping in a small set of Fagaria accessions. The ultimate aim was to create a set of correlated markers to the physiological drought-involved traits in Fragaria. Using F. vesca extensively as a characterized Rosaceae mod e-l plant species in this study can ensure the benefits of drought charact e-rization and plant-based responses in other important Rosaceae fruit species.