Durum wheat is one of the most important cereal crops in Morocco. The development of high-yielding varieties of durum wheat is one of the major goals of breeding programs in Morocco. Crop genetic diversity should play an essential role in this context. The objective of this study was to assess the genetic variation among 280 durum wheat accessions conserved at the Moroccan Gene Bank by using twelve agro-morphological characters. All accessions are planted in field conditions and arranged in augmented design. The results showed a large variability among the durum wheat accessions. Analysis of variance demonstrated significant to highly significant differentiation between the accessions for the most traits. The number of plants at the emergence stage, the biomass of the plant at the flowering stage, the chlorophyll content, and the number of spikes showed high genetic advance coupled with high heritability (broad sense). Moreover, highly significant coefficients of correlation were revealed between agro-morphological characters offering the possibility of direct or indirect selection. The first two axes of the principal component analysis explained 45% of the total variation and showed that Moroccan landraces and North American cultivars shared similar morphological characters. However, Moroccan cultivars have shown better agronomic characters. This important agro-morphological variation should be utilized as a gene pool in durum wheat improvement to develop novel varieties with high productivity and the best quality.