Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases affecting wheat. In this study, seven gene-linked markers were used to identify the presence of stripe rust resistant genes in 51 accessions of synthetic hexaploid of wheat which were found to be resistant at seedling plant stage. Molecular marker-based gene identification showed the presence of Yr5, Yr10 and Yr15 in three accessions, Yr36 in three accessions, Yr48 in seven accessions, YrR61 in four accessions, and YrTP1 in ten accessions of resistant hexaploid of wheat. These gene-linked markers were also used for the detection of genetic diversity. A total of 68 alleles were detected by these seven gene-linked markers. The mean number of allele was 11.3 alleles per locus. Genetic diversity values ranged from 0.34 to 0.93, with highest genetic diversity value of 0.93 detected for marker Xwm477. The lowest genetic diversity value was observed for marker Xbarc167. The polymorphic information content value ranged from 0.33 to 0.92 with an average of 0.54. The highest number of alleles (n= 24) were detected for marker Xwmc477. The evidence in this study on the basis of genetic diversity and presence of Yr genes in synthetic hexaploid wheat accessions will be useful in further breeding programmes.