In the present study, in order to evaluate the drought tolerance indices and their relationship with ISSR markers, 12 rapeseed genotypes were studied using a factorial experiment based on completely randomized block design under the three irrigation treatments (control and irrigation after drainage of 60 and 85% moisture content) in the greenhouse of Mohaghegh Ardabili University, Iran. Drought tolerance genotypes were evaluated by quantitative indices including MP, GMP, SSI, STI and TOL. Cultivars in all five of indices at two levels of stress exhibited significant differences. Regarding the results of the mean comparison at both levels of stress, SLMO46 was identified as the most resistant cultivar with the highest amount of MP and STI, and Karun was the most sensitive one with the highest amount of SSI index. According to the results of factor analysis, in the first level of stress, Sarigol32 and Karun were sensitive, and in the second level of stress, Talaye and Sarigol32 were sensitive as well. SLMO46 was known to be resistant to stress in both levels of stress. Phenotypic correlation of grain yield under stress and non-stress conditions was investigated in two levels of stress with 5 drought indices. In first level of stress condition, grain yield had a positive and significant correlation with mean productivity, geometric mean of productivity and stress tolerance index. In the second level of stress condition, the same correlation was observed with the difference that there was no significant correlation between drought tolerance and tolerance indices. Canonical correlation analysis was performed between drought indices and molecular markers. Five ISSR primers (5, 9, 11, 14 and 19) with the highest polymorphic percentages were used for calculation using the first factor coefficients. ISSR-PCR was used to identify some of the molecular markers associated with drought tolerance indices. A total of 106 clear and score-able loci were amplified by 18 ISSR primers, of which 60 bands (56.6%) were polymorphic. Finally, according to the results, these markers can be used in rapeseed breeding programs for drought tolerance.