The impact of osmotic stress on growth, physiology, and metabolism of winter oilseed rape(Brassica napus L.)was investigated by detailed analysis of biomass traits,hormone metabolites and osmolytes in two genetically unrelated drought-tolerant genotypes and two unrelated drought-sensitive genotypes. Seedlings were grown in vitro under controlled conditions and osmotic stress was simulated by applying a gradual treatment with polyethylene glycol(PEG6000), followed by hypo-osmotic treatment of variants used for metabolite determination. The results provide a basis for the identification of reliable selection criteria for drought resistance in oilseed rape. The in vitro cultivation system established during this study enabled effective discrimination of early osmotic stress responses between drought-resistant and -susceptible oilseed rape genotypes that also show large differences in relative seed yield under drought conditions in the field. Clear physiological and metabolic differences were observed between the drought-resistant and drought-sensitive genotypes, suggesting that osmotic adjustment is a key component of drought response in oilseed rape. Unexpected ly, however, the drought-resistant genotypes did not show typical hormonal adjustment and osmolyte accumulation suggesting that they possess alternative physiological mech anisms enabling avoidance of stress symptoms. The impact of osmotic stress on growth, physiology, and metabolism of winter oilseed rape (Brassica napus L.) was investigated by detailed analysis of biomass traits, hormone metabolites and osmolytes in two genetically unrelated drought-tolerant genotypes and two unrelated drought-sensitive genotypes . Seedlings were grown in vitro under controlled conditions and osmotic stress was simulated by applying a gradual treatment with polyethylene glycol (PEG6000), followed by hypo-osmotic treatment of variants used for metabolite determination. The results provide a basis for the identification of reliable selection criteria for drought resistance in oilseed rape. The in vitro cultivation system established during this study enabled effective discrimination of early osmotic stress responses between drought-resistant and-susceptible oilseed rape genotypes that also show large differences in relative seed yield under drought conditions in the field Clear physiological and metabolic differences w ere observed between the drought-resistant and drought-sensitive genotypes, suggesting that osmotic adjustment is a key component of drought response in oilseed rape. Unexpected ly, however, the drought-resistant genotypes did not show typical hormonal adjustment and osmolyte accumulation suggesting that they possess alternative physiological mech anisms enable avoidance of stress symptoms.