[Objective] This study aimed to investigate the effects of uniconazole on physiological and biochemical properties of roots of different sweetpotato cultivars at seedling stage. [Method] Under hydroponic conditions in a greenhouse, the effects of exogenous uniconazole on physiological and biochemical properties of roots of different sweetpotato cultivars at seedling stage were analyzed under continuous drought stress simulated by PEG-6000. [Result] Compared with normal water supply treatment (N), drought treatment (D) decreased significantly root length, root surface area, average root diameter and root volume of Jishu 21 and Jihei 1; to be specific, compared with Jishu 21, root length, root surface area and average root diameter of Jihei 1 decreased greatly, but root volume of Jihei 1 decreased slightly. SOD, POD, APX activities and MDA content of Jishu 21 and Jihei 1 increased, and root vitality of two sweetpotato cultivars was reduced; to be specific, compared with Jihei 1, Jishu 21 exhibited significantly improved SOD, POD, APX activities, slightly improved MDA content and remarkably reduced root vitality. IAA, GAs and ZR contents of Jishu 21 and Jihei 1 declined at seedling stage; ABA content of two sweetpotato cultivars was improved; to be specific, compared with Jihei 1, IAA, GAs and ZR contents of Jishu 21 were reduced slightly, while ABA content of Jishu 21 was improved slightly. Under stress conditions, uniconazole treatment increased significantly root length and root surface area of Jihei 1 and increased slightly root volume of Jihei 1 compared with Jishu 21; uniconazole treatment increased significantly SOD, POD and APX activities and root vitality of Jihei 1 and declined remarkably MDA content of Jihei 1 compared with Jishu 21; uniconazole treatment increased significantly IAA, ZR and ABA contents of Jihei 1 compared with Jishu 21. [Conclusion] Drought-tolerant sweetpotato cultivar Jishu 21 exhibits high plasma membrane integrity and root vitality by increasing activities of antioxidant enzymes under drought stress and maintaining high contents of IAA, ZR and ABA. Unicona-zole exerts greater regulatory effects on physiological and biochemical properties of drought-sensitive sweetpotato cultivar Jihei 1 under drought stress. [ABSTRACT FROM AUTHOR]