In the present investigation, 21 micro-satellite (SSR) markers were used to study the genetic diversity of 15 wheat mutant lines along with untreated check parent (HD-2867). 88 alleles were identified, the number of alleles per locus ranged from 2 to 7 with an average of 4.20 alleles per locus. It was observed that most of primers showed high polymorphism information content (PIC) ranged from 0.253 to 0.833 with an average of 0.568 per primer. Genetic similarity value ranged from 0.271 (WM-3 and WM-15) to 0.930 (WM-2 and WM-6). When comparing with parent (HD- 2867), maximum genetic similarity estimate was observed in mutant WM-4 (0.921) and lowest in mutant WM-12 (0.400). The dendogram resulting from cluster analysis revealed 2 clusters (cluster-I and II), their similarity coefficient ranged from 0.43 to 1.00. It is observed from the present study that wheat mutants showed high genetic diversity over the untreated parent check and also among themselves. High genetic diversity achieved in 15 wheat mutant lines through different doses of gamma rays (20, 25, 30kR), is a good indicator to use these mutant lines in wheat breeding programme to improve the broad genetic base and also yield.