Drought is a severe abiotic stress that affects global crop production. A drought model was created for 'Toyonoka' Fragaria × ananassa, and the effects of drought stress on contents of proline, sugars, and antioxidant enzyme activities were investigated. Strawberry transplants with identical growth were chosen for the experiments and the randomized design included four replications (10 plants per block). The experimental sets differed in the moisture level of the culture medium relative to the range of moisture content as follows: control, 70-85%; mild drought stress, 50-60%; moderate drought stress, 40-50%; and severe drought stress, 30-40%. Drought stress was imposed by limiting irrigation. Plants were sampled and physiological parameters were measured on 0, 2, 4, 6, 8, and 10 days after the commencement of drought stress. The water potential of strawberry leaves decreased in the plants under mild, moderate, and severe stress during the course of the water stress treatment and exhibited a significant difference from the control. Strawberry leaves subjected to drought stress had higher accumulation of proline, sugars, and malondialdehyde, and higher activities of superoxide dismutase, peroxidase, and catalase than leaves of control plants. Malondialdehyde levels increased in parallel with the severity and duration of drought stress. By contrast, antioxidant enzyme activity displayed dynamic responses to drought stress, first increasing and subsequently decreasing as the severity and duration of drought stress increased. These results suggest that strawberry plants respond to drought stress by altering the activities of antioxidant enzymes and the levels of osmotically active metabolites. These biochemical response changes may confer adaptation to drought stress and improve the capacity of plants to withstand water-deficit conditions.