In order to evaluate the response of bread wheat genotypes to callus induction and in vitro drought stress, an investigation was carried out as a factorial experiment with a completely randomized design and five replications. The results of analysis of variance indicated significant differences between the entries and stress levels for callus relative growth (CRG), callus relative growth rate (CRGR), callus growth rate (CGR), percentage of callus chlorosis (PCCH) and percentage of callus water content (PCWC) indicating the presence of genetic variability, different responses of genotypes to different drought intensities and in vitro selection of drought-tolerant genotypes. Mean comparisons between genotypes revealed that maximum CRG, CGR, CRGR, PCWG, PCCH and INTOL were attributed to genotypes 5, 16, 17, 2, 3 and 20 (drought tolerant), respectively. Graphic observation exhibited that indices of drought tolerance decreased with increase of PEG concentrations. Cluster analysis of genotypes (Ward's method) based on CRG, CGR, CRGR, PCWG, PCCH and INTOL and subsequent discriminant analysis for confirming the number of clusters, grouped the genotypes into four different clusters. The first group included genotypes 1, 3, 6, 7, 12, 13 and 18, the second group included genotypes 2, 4, 8, 9, 10 and 11 and the third group consisted of genotypes 14, 16, 17, 19 and 20, while the genotype 15 formed the fourth group. Superior genotypes 2, 5 and 16 showed drought tolerance at the callus culture level together with their high potential for callus induction leaded us to the conclusion that a hybridization breeding program using these superior plant materials supplemented with in vitro selection for drought tolerance might be beneficial for improvement drought tolerance in bread whea