A total of 37 allele was amplified with each primer thus produced on an average 7.4 polymorphic bands and The number of alleles per locus ranged from 5 to 12. The polymorphism index content (PIC) value varied from 0.675 to 0.868. ISSR primers performed well in the detection of in wheat genetic diversity can be recommended for future wheat improvement program. Genomic DNA was extracted from thirty three wheat genotypes using CTAB procedure and amplified using five ISSR primers. Matrix similarity of genotypes was calculated by using NTSYSpc.2.1 with Sanh-clustering using the UPGMA (Unweighted Paired Group Method Using Arithmetic Averages) method. The prime aim of this study were to determination of the genetic polymorphism, genetic diversity, selection promising diverse progenitor, identification of putative and informative markers can be recommended for future crop improvement program.