Mutation is a resource of generating genetic diversity in crop plant for breeding as well as genetic analysis. Ethyl Methane Sulphonate (EMS) is frequently applied chemical agents in plants. The present study was completed to investigate the mutagenic effects of different concentration of EMS on yield contributing traits of super basmati. The seeds of rice cultivar (Super basmati) were mutagenized with different doses from 0-2% v/v of EMS with difference of 0.25% for determination LD50. The treated and non-treated plants were observed under different agronomic parameters. A total of 48 putative EMS mutants of super basmati were selected randomly to analyze genetic diversity using 25 SSR primers. These markers were located on twelve chromosomes of rice. SSRs analysis revealed that abroad level of genetic diversity was existed among mutants of super basmati. A sum of 91 alleles was identi ed of which 82 alleles were originated to be polymorphic and the rest of nine alleles were discovered as monomorphic. The range of allele number was 2-10 with mean of 3.64 alleles/locus. The value of polymorphic information content (PIC) was from 0.039 (RM5) to 0.878 (RM44) with mean of 0.439 for each locus. All mutants differentiated from each other in more than two set of primers due to presence or absence of unique bands on chromosomes at de nite base pair. The size base pair range was 75-1000bp. Dendrogram located mutant indices into four major groups. Phylogenic analyses exposed 40-96% similarity. It is concluded that EMS induced genetic variability and SSRs markers (RM44, RM154, RM1, RM252, RM334, RM487, RM110 and RM257) could be employed for the selection of rice mutants throughout molecular breeding program.