To reveal the effects of carbon sources on callus induction and proliferation, we studied comparative analyses of 30.0 g/L sucrose, glucose and maltose on callus induction and proliferation by using pedicel as explants in Agapanthus praecox ssp. orientalis. Physiological indicators of callus proliferation stage were determined, and the correlation coefficients of cell proliferation and physiological indexes were evaluated. Furthermore, the effectiveness of some physiological indicators on callus proliferation was confirmed. Here are the results. (1) The callus induction rate was 86.00%, 72.00% and 59.67% with sucrose, glucose and maltose treatment, respectively. The callus induction rate of sucrose treatment significantly increased by 19.44% and 44.13% compared with glucose and maltose respectively (P < 0.05). Meanwhile, the callus size of sucrose treatment significantly increased by 22.44% and 90.09% compared with glucose and maltose respectively (P < 0.05). In callus proliferation stage, sucrose can maintain both proliferation efficiency and cell viability. However, glucose treatment induced higher cell proliferation rate and poor cell viability, and maltose treatment induced slow cell proliferation rate and vigorous cell viability. Carbon sources conversion treatment indicated that callus cell mass size and cell state decreased sharply, when callus transferred from sucrose to glucose medium. However, the proliferation efficiencies of both from sucrose to sucrose and from sucrose to maltose were well. (2) Carbon sources significantly regulated sugar metabolism, endogenous hormone metabolism and oxidative stress balance in the process of callus proliferation. (3) Starch and glucose were the principal saccharides components of callus, and the contents of starch and maltose were highly correlated with cell mass proliferation efficiency. The combination of sucrose and maltose optimized callus proliferation, the cell mass color was bright yellow, and cell activity was vigorous. (4) Contents of binding IAA, GA₄, and CTK were correlated with callus cell mass size. The addition of 1.0 mg·L^-1 6BA considerably promoted cell proliferation efficiency (P < 0.05). (5) ROS activity was negatively correlated with POD and CAT activities, and POD activity was negatively correlated with H₂O₂ content (P < 0.05). The POD and CAT activities were extremely significant positive correlated (P < 0.01). (6) The results of validation and optimization experiments showed that the addition of maltose and 6BA in the culture medium effectively promoted callus proliferation in A. praecox, in which maltose maintained and improved the cell activity, while 6BA mainly promoted cell proliferation. In conclusion, sucrose was the most appropriate carbon source for callus induction and proliferation in A. praecox. Sucrose and maltose combination improved cell viability, while picloram and 6BA combination accelerated callus proliferation. The optimized medium for callus proliferation was MS + 0.5 mg/L PIC + 1.5 mg/L 6BA + 15.0 g/L sucrose +15.0 g/L maltose + 7.0 g/L agar.