**4. Conclusions**

This study aimed to develop M5 mutant lines of spring wheat by 100-and 200-Gy gamma irradiation treatment and identify genetic variability in grain nutritional value including grain protein content, grain Fe and Zn concentrations, and yield-associated traits such as grain number and weight per main spike, grain weight per plant, and 1000-grain weight and grain morphometric parameters, namely grain area, length, and width and M5 mutant lines in comparison to the cv. Eritrospermum-35 parent. We identified mutant lines with high Fe and Zn concentrations and grain protein content as those which have 1.6–3.4 and 1.4–2.9 times more as well as 3.7–16.9% than target parameters of parent variety, respectively. Several mutant lines showed significantly simultaneous increase of metals concentrations. The positive correlation of grain protein content with grain width suggests that they are genetically related. This relation could be a good candidate for evaluating genotypes for improvement of grain protein content. Wheat grain can be biofortified by micronutrients without negative impact on crop productivity using new mutant lines. Mutation breeding can significantly contribute to human health malnutrition and improve nutrition quality diet.

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