Analysis of genetic diversity provides helpful information necessary to develop the breeding and conservation strategies of crops. In this study, the genetic diversity and population structure of 90 durum wheat genotypes maintained at Sararud Dryland Agricultural Research Institute, Kermanshah, Iran, were evaluated by using 23 gene-specific markers (functional markers, FMs) encoding high and low molecular weight glutenin and gliadin alleles. Results showed that 12 out of the 23 FMs used were polymorphic and amplified 52 polymorphic loci. Primer Ax2 ⃰ had the highest discriminatory power. The population structure analysis classified the durum wheat collection into four populations. On average, population 4, consisting of 8 genotypes, had the highest allele number as well as genetic variation. Analysis of molecular variance indicated that 82% of the total variation was distributed among populations. The diversity among populations and gene flow were 0.14 and 3.03, respectively. The Jaccard distance coefficient revealed that genetic dissimilarities ranged from 0.031 between G62 and G65 to 0.725 between G36 and G51. Neighbor-joining method clustered individuals into six main groups. Results showed a remarkable level of genetic diversity among studied durum wheat genotypes which can be of interest for future breeding programs.