Narrow genetic variability may lead to genetic vulnerability of field crops against biotic and abiotic stresses which can cause yield reduction. In this study a set of 37 wheat microsatellite markers linked with identified QTLs for salinity tolerance were used for the assessment of genetic diversity for salinity in 30 promising lines of hexaploid bread wheat (Triticum aestivum L.). A total of 438 alleles were detected with an average allele number of 11.84 per locus using 37 microsatellite markers. The number of alleles per locus ranged from two to twenty, the maximum number of alleles was observed at Xgwm312. Gene diversity statistic for 37 microsatellite loci was varied from 0.66 to 0.94 and also polymorphic information content value was varied from 0.64 to 0.93 for Xgwm445 and Xgwm312 respectively. Result showed Xgwm312 SSR marker with the highest PIC value was distinguished as the best marker for genetic diversity analysis to improve of salinity tolerance. Obtained dendrogram by UPGMA method categorized genotypes in to 3 different groups, which had different reaction to salinity. A wide range of genomic diversity was observed among all the genotypes. Principal Coordinates Analysis (PCoA) also confirmed this pattern of genetic diversity, proving them can be use as the prime candidates in order to improve of salinity tolerance in breeding programs of wheat. The present study also indicates that microsatellite markers permit the fast and high throughput fingerprinting of numbers of genotypes from a germplasm collection in order to assess genetic diversity.