Drought stress was evaluated with polyethylene glycol 6000 (PEG 6000) treatment in Dunaliella salina, a microalga known for its great ability to withstand salinities of more than 30%. The aim was to explore the acclimation mechanisms used by the microalga to regulate its growth and physiology during coping with drought stress. The microalga was subjected to culture mediums containing 2 and 5% PEG for 25 days and was compared with a control culture medium. Significant decrease in growth parameters such as specific growth rate, biomass and number of divisions per day was demonstrated in PEG-treated algae. During PEG treatment, chlorophylls slightly increased, while β-carotene and total protein were not affected. Osmolytes, as well as carbohydrates, were found to be significantly higher in PEG-treated algae than in control. Increased catalase and ascorbate peroxidase activities were proportionally related to PEG concentrations in the cultures. The PEG-treated cells accumulated a considerable amount of hydrogen peroxide and malondialdehyde, especially at higher PEG concentrations. Electrolyte leakage increased, regardless of the PEG concentrations applied, while DNA fragmentation was not observed after 25 days of treating with PEG. It was concluded that Dunaliella cells could respond to the drought stress, probably by using a higher accumulation of a range of osmolytes and also more stimulation of the antioxidant enzymatic system.