In this study, we selected 180 accessions from different wild relatives of wheat (Aegilops–Triticum species) and tested them in the presence of a tolerant and a sensitive control variety under well-watered and drought-stressed conditions. The results of analysis of variance for dry masses and all measured root traits showed that water regimes, accession and species main effects were highly significant. Drought stress significantly declined shoot dry mass (59.42%), root length (37.85%) and the total number of branch roots (36.25%), but increased the root-to-shoot ratio (75.00%), specific root length (64.19%) and root tissue density (29.46%). Principal component analysis for 182 individuals and 12 species groups identified two components that explained 75.67 and 82.39% of the total variation in dry mass and root traits under drought-stressed conditions, respectively. Taking together, our results identified 12 accessions with superior tolerance to drought stress. Remarkably, four species of wild relatives – Ae. cylindrica (DC genome), Ae. neglecta (UM genome), Ae. speltoides (B genome) and Ae. tauschii (D genome) – responded well to drought stress. The potential of these species could be used for molecular analysis such as marker assisted selection and gene mapping, ultimately aimed at breeding for root traits with improved adaptation to drought environments.