The present study was designed to evaluate the effect of ¹²C⁶⁺ ion beam (10~80 Gy) on biological changes of wheat seedlings. Reactive oxygen species (ROS)-related biomarkers and the quantification of plant survival and growth were examined at 10 day after carbon ions irradiation (LET: 30.8 keV/μm). The results showed that heavy ions obviously enhanced ROSs reflected by the production of O₂^- and H₂O₂ as well as TBARS, and treatment with 20 Gy achieved the peak value, suggesting that higher mutagenic potential may occur at 20 Gy. Simultaneously, increase of SOD activity was induced by heavy ions to counteract ROS accumulation. On the other hand, higher doses at 40 and 80 Gy inhibited wheat growth and survival in comparison with the control, and reversely lower doses at 10 or 20 Gy stimulated wheat growth and survival. In conclusion, the above observations imply that a dose range of 20~40 Gy is likely promised for wheat mutation breeding.