Abstract: To explore the regulatory role of exogenous glycine betaine (GB) on the growth of creeping bentgrass (Agrostis stolonifera) under drought stress and rehydration, creeping bentgrass sods were transplanted into a greenhouse and grown for 50 days before spraying with GB using 4 different concentrations (0, 50, 100 and 200 mmol/L). Artificial drought treatment and rehydration were applied 3 days after GB application. Turf quality, relative water content, cell membrane permeability, chlorophyll content, free-proline content, soluble protein content, malondialdehyde (MDA) content, and the activity of antioxidant enzymes; superoxide dismutase (SOD); catalase (CAT); and peroxidase (POD) were measured. The results showed that turf quality, relative water content and chlorophyll content were decreased by drought stress while cell membrane permeability, free-proline content, soluble protein content, MDA content and SOD, CAT, and POD activity were increased. However, application of GB significantly reduced the decline of turf quality, relative water content, and chlorophyll content under drought stress. Application of GB also increased the free-proline content, soluble protein content, SOD and CAT activity but reduced cell membrane permeability and MDA content compared to the control. Recovery after rehydration was also improved by GB application. The results suggested that exogenous GB could enhance the drought tolerance of creeping bentgrass under drought stress and the optimum concentration was100 mmol/L.