Nickel (Ni) is known as a plant micronutrient and serves as a component of many significant enzymes, however, it can be extremely toxic to plants when present in excess concentration. Scientists are looking for natural compounds that can influence the development processes of plants. Therefore, it was decided to use proline as a protective agent against Ni toxicity. Proline (Pro) is a popularly known osmoprotectant to regulate the biomass and developmental processes of plants under a variety of environmental stresses, but its role in the modulation of Ni-induced toxicity in wheat is very little explored. This investigation indicated the role of exogenously applied proline (10 mM) on two wheat varieties (V1 = Punjab-11, V2 = Ghazi-11) exposed to Ni (100 mg/kg) stress. Proline mediated a positive rejoinder on morphological, photosynthetic indices, antioxidant enzymes, oxidative stress markers, ion uptake were analyzed with and without Ni stress. Proline alone and in combination with Ni improved the growth, photosynthetic performance, and antioxidant capacity of wheat plants. However, Ni application alone exhibited strong oxidative damage through increased H2O2 (V1 = 28.96, V2 = 55.20) accumulation, lipid peroxidation (V1 = 26.09, V2 = 38.26%), and reduced translocation of macronutrients from root to shoot. Application of Pro to Ni-stressed wheat plants enhanced actions of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and total soluble protein (TSP) contents by 45.70, 44.06, 43.40, and 25.11% in V1, and 39.32, 46.46, 42.22, 55.29% in V2, compared to control plants. The upregulation of antioxidant enzymes, proline accumulation, and uptake of essential mineral ions has maintained the equilibrium of Ni in both wheat cultivars, indicating Ni detoxification. This trial insight into an awareness that foliar application of proline can be utilized as a potent biochemical method in mitigating Ni-induced stress and might serve as a strong remedial technique for the decontamination of polluted soil particularly with metals.