Watermelon variety 'Liaoyin No.1' were treated at the age of three leaves by spraying brassinolide in the concentration of 0(CK),0.01,0.05,0.10,0.50,1.00 mg/L,and then were stressed under temperature(10±1)℃ and light 100 μmol·m-2·s-1.In order to improve chilling resistance of watermelon seedlings,the physiological and biochemical characteristics of watermelon seedlings were measured to screen a optimum concentration of brassinolide(BR).The results showed that the best seedlings growth was observed under the treatment 0.10 mg/L of BR,which could reduce the chilling injury index significantly,increase the activities of superoxide dismutase(SOD),peroxidase(POD) and catalase(CAT) and reduce the content of malondialdehyde(MDA) and superoxide anion radical.