Abstract: Glycine betaine (GB) is an important compatible solute that is associated with stress tolerance in plants under low-temperature stress. To investigate the effect of exogenous GB on the physiological indexes of Zoysia japonica under low-temperature stress, GB at 0, 50, 100, and 150 mmol/L was applied as a foliar spray to ‘Qingdao’, a Z. japonica cultivar, before transferring the plants to growth chambers. The plants were grown under average temperatures of 8℃/2℃ (day/night) for 28 days in the low-temperature treatment, and under average temperatures of 28℃/24℃ (day/night) in the control. Exogenous GB alleviated the loss of turf quality and chlorophyll content, and reduced the increase in electrolyte leakage and malondialdehyde content in Z. japonica under low-temperature stress. Exogenous GB increased the activities of superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase, leading to lower H₂O₂ and O2·- contents. Also, exogenous GB enhanced the osmotic adjustment capacity of the plants by increasing the concentrations of proline and soluble protein. These results indicated that exogenous GB enhanced the resistance of Z. japonica to low-temperature stress, and 100 mmol/L exogenous GB was the most effective concentration.