The aim of this research is to assess the effectiveness of gamma irradiation in developing genetic variability in dragon fruit by influencing biochemical attributes. Seeds of dragon fruit were exposed to 100, 200, 300, 400 and 500Gy gamma-radiation using Co-60 source. Highest germinability, mean germination time, mean daily germination, vigour index and growth rate were observed in untreated seeds, however these traits were relatively less affected at lower irradiation level (up to 200Gy). An irradiation level of >= 400Gy severely affected germination traits and seedling growth and consequently 441Gy irradiation level was considered as the lethal dose (LD50) for dragon fruit. Photosynthetic pigments, protein content and nutrient content were highest in untreated seedlings and moderately high up to 200Gy. Findings demonstrated that the concentration of biochemical markers for abiotic stress tolerance viz. proline, phenol, flavonoid and antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase), were significantly high at 200Gy irradiation level. Hence an irradiation dose of 200Gy may be considered as an optimal for induced mutagenesis for abiotic stress tolerance in dragon fruit with the least possible unintended damage to seedling growth traits. The findings provide valuable insights into the efficient utilization of gamma-irradiation in expediting the development of abiotic stress-tolerant mutant lines of dragon fruit.