Acclimation is a process of adjustment to gradual environmental change that enables plants to survive further stress by triggering some tolerance mechanism possibly involving ABA, ROS and oxidative metabolism. Here we have studied acclimation responses in terms of the performances with regard to physiological (growth and relative water content) and biochemical (chlorophyll, carotenoids, protein, malondialdehyde, sugar content) attributes, ABA production and stomatal sensitivity to exogenous ABA, extracellular ROS production and activation of antioxidant system. Our study reveals that repeated exposure to short-term mild water stress simulated by polyethylene glycol (PEG-6000) induces acclimation in mung bean (Vigna radiata L. Wilczek) seedlings. Acclimation induced tolerance was associated with reduced leaf size and enhanced root growth, accumulation of soluble sugar as osmoprotectant, maintenance of water potential, lessening of membrane damage as indicated by lower MDA content. Acclimated mung bean seedlings have shown greater degree of tolerance through increased production of and enhanced sensitivity to ABA (as reflected by faster stomatal closure), enhanced production of extracellular O2.- and H2O2 and the elevated activities of antioxidative enzymes to control the oxidative burst. Taken together, the results convey that acclimated seedlings minimize osmotic stress-induced damage through a possible network of ABA, ROS and antioxidants.