Sorghum (Sorghum bicolor (L.) Moench) is a climate-resilient cereal vital for food security, especially in arid and semi-arid regions. Understanding the molecular characterization of sorghum mutant genotypes is crucial for crop improvement, yet their genetic diversity and population structure remain poorly understood. This study assessed 190 randomly selected mutant genotypes from 2000 M2 lines generated via Ethyl Methanesulfonate (EMS) treatment, alongside two parental lines, using 10 Simple Sequence Repeat (SSR) markers. Key diversity metrics, including polymorphic information content (PIC), genetic diversity (GD), and allelic richness (Ar), were analyzed. The SSR analysis revealed significant polymorphism, with mean values of 0.84 (PIC), 0.83 (GD), and 6.41 (Ar), indicating substantial genetic variation. Analysis of Molecular Variance (AMOVA) showed that 97 % of genetic variation occurred among individuals, with only 2 % and 1 % attributed to population and withinindividual variations, respectively. The high within-population variation suggests extensive genetic diversity due to mutagenesis and selection. Despite this, moderate population divergence was observed, indicating genetic relatedness among groups. Cluster analysis identified two distinct genetic groups, with most clusters containing mutants from both parental lines, reflecting shared ancestry. Several mutants with high genetic diversity were identified as promising candidates for multi-location agronomic trials, particularly for drought resilience and yield stability. These findings highlight the potential of these genotypes for breeding programs aimed at enhancing sorghum resilience and productivity.