Vanadium (V) as minor concentration is present in various plants and extensively found in soils. The current study was established to assess the response of rice seedlings to different V concentrations and also investigated its toxic effect on growth, photosynthetic assimilation, relative chlorophyll content, SPAD index, ion leakage, enzyme activities, hydrogen peroxide (H 2 O 2), and cell death. The rice seeds were sown in Petri dishes for 8 days, and after that, rice seedlings were grown hydroponically in a climate-controlled growth chamber. After 15 days of V-treatment, antioxidant enzyme activities, H 2 O 2 , protein contents, photosynthetic assimilation, relative chlorophyll content, and cell death were determined by utilizing the Spectrophotometer (Lambda 25 UV/VIS Spectrophotometer), and V accumulation (roots and shoots) was determined by GFAAS (GTA 120). The obtained results showed that all V concentrations significantly decreased the biomass (dry and fresh) and root growth as a result of the reduction in total root length, root tips, root fork, root surface area, and root crossing, and V was more accumulated in roots than shoots. Besides this, enzymatic activities were significantly enhanced under V stress. The findings also confirmed that seedling exposed to V stress had lower tolerance indices, photosynthetic activity, and protein contents while the ion leakage was consistently increased by increasing the V concentrations. The viability of plant cells severely damaged in response to high V stress, and H 2 O 2 induction might be responsible for cell death. Generally, all V doses had a drastic effect on enzyme activities and caused cell death of rice plans. Moreover, the current study demonstrated that V ≥ 35 mg L −1 caused damaging effects on rice plants.