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During the present study, we successfully developed a mutant population of wheat by utilizing the genetic background of a high-yielding wheat variety, namely DPW 621-50. To accomplish this, a total of 42,858 seeds from DPW 621-50 were subjected to treatment with 0.7% EMS, resulting in the production of over 10,000 M2seeds. Each of these seeds underwent individual screening to identify any altered phenotypes, a process that extended up to the M4generation. Significant variations in phenotypic traits, including plant height, tiller number, spike length, spikelet number, and peduncle length, were consistently observed within the population across successive generations. Additionally, a comprehensive study was conducted to assess the levels of four essential micronutrients—namely, Fe, Zn, Ca, and Cu—in the whole grains of these mutants. Among the 423 mutant lines analysed in this study, as many as 94 plants exhibited elevated levels of Ca, Cu, Fe, and Zn, even after accounting for concentration effects. Notably, mutant lines such as DFW-21 (with a Ca content of 206.36 ppm) and Bus-2-P-2 (with a Ca content of 204.63 ppm) demonstrated the highest Ca content. Similarly, M-9-P-4 and DWF-47 exhibited the highest Cu content (8.04 ppm and 7.82 ppm, respectively), while M-152-P-1 showed the highest Fe content (89.1 ppm). Furthermore, M-152-P-1 and M-9-P-4 exhibited the highest Zn content (60.78 ppm and 53.34 ppm, respectively). These findings underscore the potential of these mutant lines for inclusion in wheat biofortification programs, offering a promising approach to address the global challenge of malnutrition. Moreover, the identification of these lines opens doors for enhancing not only nutrient content but also other critical agronomic traits.