【Objective】 To better understand how 24-epibrassinolide (EBR) affects the development and physiological characteristics of ginkgo, different concentrations were sprayed onto ginkgo leaves, laying a theoretical basis upon which the oriented cultivation of ginkgo can be applied. 【Method】 Ginkgo leaves were placed in a randomized block design and treated with different concentrations of EBR (0-2.0 mg/L). The growth index, photosynthetic characteristics, and hormone content of EBR-treated annual seedlings were measured. 【Result】 The growth of ginkgo seedlings differed significantly following EBR treatment (P <0.05), with 1.0 mg/L EBR leading to increases of 23.6%, 52.6%, 140.2%, 87.7% and 69.1% in the maximum leaf length, leaf area, plant height, leaf weight and total weight, respectively, as compared to the control. Treatment with 1.5 mg/L (BR3) and 2.0 mg/L (BR4) led to increases of 40.2% and 41.5%, respectively, in the net photosynthetic rate as compared to the control. The maximum content of the endogenous hormones dihydrozein, indolepropionic acid, gibberellin, and abscisic acid and the minimum auxin content was observed under 1.5 mg/L EBR. The content of endogenous brassinolide was significantly positively correlated with the contents of methyl jasmonic and indolepropionic acid and negatively correlated with zeatin (R > 0.5, P < 0.05). 【Conclusion】 The results indicate that the exogenous spraying of ginkgo with EBR is an effective agronomic measure that could improve the net photosynthetic rate of gingko, while also regulating the content of endogenous hormones in the plant, thus promoting stem elongation, leaf area expansion, and leaf dry weight accumulation.