[Objective] This study aimed to investigate the effects of drought stress on osmotic regulation substances of Catalpa bungei clones. [Method] Potting experimental was carried out with plastic film sealing method, to differentiate the degrees of drought stress based on changes in soil water content and leaf water potential and investigate variations in contents of osmotic regulation substances(free proline,soluble sugar and soluble protein) in leaves of five Catalpa bungei clones under different degrees of drought stresses. [Result] According to changes in soil water content and leaf water potential, the whole process of drought stress was divided into normal level(CK, with soil water content of 97.49% and leaf water potential of-0.54MPa), light drought(LD with soil water content of 59.96% and leaf water potential of-1.28 MPa), mediate drought(MD with soil water content of 34.19% and leaf water potential of-2.32 MPa) and severe drought(SD with soil water content of 14.52%and leaf water potential of-2.99 MPa). The soil water content and leaf water potential of five Catalpa bungei clones reached the highest correlation in exponential fitting, with an average R2of 0.989 3(P0.001). The free proline content in leaves of five Catalpa bungei clones increased rapidly with the increasing degree of drought stress(P 0.001). To be specific, free proline contents of Catalpa bungei clones015-1 and 7080 were 34.39 and 33.41 times of the normal level under severe drought conditions, which reached an extremely significant level(P0.001); the free proline content of Catalpa bungei clone 1-3 rapidly increased to(855.46±227.52) μg/g Fw under light drought conditions. The soluble protein content in leaves showed different variation trends. To be specific, the soluble protein content of Catalpa bungei clone 7080 was the lowest at various drought stages and reached(1.644 ±0.137)mg/g Fw under normal conditions; the soluble protein content of Catalpa bungei clone 1-3 was relatively high under norm