In the present study deals with the Oryza sativa .L was selected for callus induction and regeneration in in vitro condition and also the physiological and biochemical studies were carried under the different salinity levels. The different concentration of 2, 4-D were used to induce callus in the mature seeds and the callus was developed from the region of the seeds and was visible within 7-10 days. The salt stress significantly influenced the germination percentage was reduced gradually with increasing salt stress from 0 to 300 mM NaCl. The rate of proline accumulation was observed much higher (205.4 mg/g) at 300 mM NaCl stress as compared to control (70.15). A continuous decrease in protein content with increase in salt stress was observed, where around 64% reduction was observed in plants under 300 mM NaCl stress as compared to the control plants. The starch content was higher in control (617.2) than sample (345.5) with salinity stress. Among this study investigated that the overall showed better tolerance to salt stress with a lesser extent of antagonistic effect of NaCl on germination and biomass production at seedling stage.