Abstract
Wheat, a staple crop cultivated for over 8000 years, sustains more than 2.5 billion people globally, as a major source of carbohydrate, protein, fiber, and essential nutrients. Colored wheat, enriched with dietary fiber and antioxidants, offers valuable genetic resources for developing functional wheat varieties. Herein, a mutant pool of 1069 colored wheat lines was developed through gamma-ray irradiation to enhance genetic diversity. Mutant lines were classified into 10 groups based on seed color parameters (L*, a*, and b*), which were measured using the Hunter Lab system. K-means clustering categorized the mutant lines, and four representative lines from each group were analyzed for agronomic traits (plant height, spike length, thousand-seed weight, and kernels per spike) and antioxidant properties (radical-scavenging activity, ferric reducing antioxidant power, and total antioxidant capacity). Principal-component analysis revealed distinct clustering patterns, indicating associations between seed color, agronomic traits, and antioxidant activity. Darker seed color groups exhibited 3-16% higher levels of bioactive compounds and 10-18% higher antioxidant activities, whereas lighter groups showed 8-42% lower functional potential compared to the control wheat. These findings highlight the potential of mutation breeding in generating phenotypic diversity and developing wheat varieties with improved functional traits and bioactive compound content.