Characterization of germplasm by agro-morphological, biochemical and DNA-markers provide powerful tools to precise germplasm identification, quantify the genetic diversity and to estimate the phylogenetic relationship among accessions in many crop species. The objective of the present study was to characterize a subset of 23 Moroccan relevant durum wheat (Triticum turgidum L. var. durum) accessions selected from the National Gene Bank of Morocco for their genetic diversity using agromorphological and microsatellite markers, and to measure the genetic distance among these accessions using UPGMA cluster analysis. Durum wheat accessions were planted under field condition and their agro-morphological characters such as days to emergence, days to tillering, days to booting, days to head emergence, days to flowering, days to physiological maturity, plant height, and thousands kernel weight were recorded. The same set of accessions was also analyzed for polymorphism with 7 genomic microsatellite markers. The results indicated that the evaluated germplasm comprises of useful variation for future breeding activities. Furthermore, the cluster analysis based on microsatellite data showed the closest correlation with the groupings of particular genotypes based on agro-morphological characters. The authors results suggest that the characterization based on agro-morphological traits and genotypic markers will be a useful tool to the breeders to choose genotypes with appropriate diversity.