Sustainable production and improved genetic gains can be achieved by broadening the genetic window of elite wheat germplasm. Here, we induced mutations in two spring wheat varieties, viz., NN-Gandum-1 (NN-1) and Punjab-11 (Pb-11), by exposing their seeds to ethyl methane sulfonate (EMS) and γ-rays, respectively. We characterized >3500 lines of each NN-1 and Pb-11 derived population in three consecutive generations, viz., M₅, M₆, and M₇, for important traits, e.g., plant height, heading date, spike morphology and rust resistance. We observed significant genetic variation and correlations in both populations for all investigated traits. We observed differences in terms of number of mutants between NN-1 (22.76%) and Pb-11 (26.18%) which could be ascribed to the genotype-by-mutagen interaction. High broad-sense heritability (H²) estimates, that are vital for higher genetic gains, were observed for all of the investigated traits in both populations (H² = 0.69–0.91 in NN-1 and 0.84–0.98 in Pb-11). Particularly, to breed for rust resistance, we selected a subset (n = 239) of M₇ lines that also showed phenotypic variation for other traits. Our studies (1) show the relevance to artificial mutagenesis to create genetic variation in elite germplasm for their immediate use in current breeding programs, and (2) provide material for downstream identification of genes associated with traits of high agronomic importance.