Calluses of two autotetraploid rice (DPR (4) and Zijing (4)) were implanted with 30 KeV N+ beams at doses ranging from 1.0×1015 to 9.0×1015 ions cm-2. The differentiation rates of calluses, the concentration of malondialdehyde (MDA), the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), and the change of esterase (EST) spectrum were measured and analyzed on the calluses stage. The results showed that differentiation rates of calluses under 3.0×1015 ions cm-2 ((46.12±2.01)%) were higher than the other treatment groups in DPR (4), and those of Zijing (4) were similar under the doses of 1.0×1015, 3.0×1015 and 5.0×1015 ions cm-2 ((40.55±2.74), (37.77±3.04) and (34.56±2.65)%) and were higher than that of 7.0×1015 and 9.0×1015 ions cm-2. When the doses were 3.0×1015 and 5.0×1015 ions cm-2, the activities of SOD, POD and CAT were notably enhanced, and the accumulation of MDA content was markedly alleviated in the implanted calluses of two materials, as well as the activity of EST was increased by the additional isforms DPR (4). Whereas the anti-oxidative systems and enzymes spectrum of EST were destroyed seriously, the accumulation of MDA was significantly aggravated, also the differentiation rate of calluses reduced sharply and almost closed to zero when the implantation doses were 7.0×1015 and 9.0×1015 ions cm-2. It suggested that the proper dose of ion implantation effectively maintained the normal membrane structure and metabolism, and it would be also a feasible approach for autotetraploid rice to improve its genetic characteristics at calluses stage. Additionally, the differences of the radiation resistance between DPR (4) and Zijing (4) could be related to the different genotypes.