The persistence of xenobiotic substances in the environment, including propellants, explosives, and pyrotechnic compositions, has become a global threat. Previous research has shown that nitroaromatics have an adverse influence on plant growth and development. Therefore, this study aims to examine how 2,4-dinitrotoluene (DNT) affects Oryza sativa physiologically and biochemically. The detrimental effects of DNT on O. sativa growth parameters (germination, viability, and electrolyte leakage) were seen at various concentrations (0, 0.025, 0.05, and 0.1%, w/v). It was discovered that these growth characteristics were sensitive to DNT, and that increases in DNT concentrations had detrimental effects. To further comprehend their harmful effects, a variety of biochemical (reactive oxygen species; ROS, malondialdehyde; MDA) and molecular investigations were carried out. When DNT concentrations increased, ROS and MDA levels significantly increased (superoxide radicals: 1.33-3.12, hydrogen peroxide: 1.54-1.89, and MDA: 2.8-5.65 times). Additionally, the plant system contains a range of enzymatic antioxidants to combat ROS and the harmful reactions they cause. The level of enzymatic antioxidants (superoxide dismutase, catalase, and ascorbate peroxidase) was measured spectrophotometrically in the current study, and the results show that exposure to DNT has a major impact on antioxidant levels. In conclusion, DNT exposure causes phytotoxic effects on O. sativa, which can be seen by evaluating physiological and biochemical processes. The degree of damage was found to be strongly associated with the DNT content.