Glycine max (L.) Merr.production is affected by a number of biotic and abiotic stresses. In order to overcome these challenges, development of transgenic plants, production of high-quality varieties and secondary metabolites, preservation of rare plants and protection of germplasms is of prime importance. In this regard, proper regeneration system for plant growth is needed to overcome dormancy and low germination issues by using appropriate growth hormones. In the proposed study, Glycine max (Variety Swat84) was assessed to investigate salt tolerance on the basis of growth, yield, physiological and biochemical responses. Seeds were germinated on MS medium with different concentrations of hormones for plantlet formation. Excellent callus was formed using 2, 4-D (2, 4-dichlorophenoxyacetic acid) at 4.0 mg L-1 concentration. Significant induction and elongation of shoot were recorded using a combination of BAP (6Benzylaminopurine) +KIN (Kinetin) at 2.0 mg L-1 + 1.0 mg L-1 respectively while maximum root induction and elongation were recorded using IBA (Indole-3-butryic acid) at 0.5 mg L-1 concentration. For the salt tolerance experiment, 0.10 g of callus was shifted to MS medium supplemented with NaCl (sodium chloride) at concentrations of 0, 50, 100, 150 and 200 mM.Callus formation showed considerable decrease at all salt concentrations as compared to control. Under saline conditions, stunted shoot growth, leaf chlorosis and defoliation occurred with increasing salt stress. The root length also decreased significantly at all salt treatments from 50 mM NaCl to 200 mM NaCl.The screening of selected clones were performed by using growth rate in term of fresh and dry weight as well as fresh and dry weight ratio of callus, shoot and root. Membrane thermo stability (EC), lipid peroxidation (MDA) and Hydrogen peroxide (H2O2) assays were performed to assess the stress induced damages. Salt stress significantly showed high electrolyte leakage of membranes and MDA contents both in callus and shoot. While, H2O2 intensity in callus showed an increase at 100 mM concentration and in shoot, an increase in H2O2 intensity was observed at 50 mM.Whereas, maximum proline was found at 100 mM NaCl concentration in both callus and shoot. Regarding total soluble protein, an increase at 200 mM concentration was observed in callus while an increase was observed at 100 mM NaCl in shoot.Furthetmore,SDS-PAGE analysis revealed that in both callus and shoot, the band intensity decreased with increasing salt stress level, but at 100 mM concentration, a 75kDa band was differentially expressed and all bands showed almost lowest level of intensity at NaCl 200 mM. From the results of present analysis, it can be concluded that salt stress has a destructive effecton the plants development, growth, physiology and yield.The intense effects of salinity can be overcome by using salt tolerant cell lines and maintain sustained growth on medium supplemented with NaCl when compared to salt sensitive cell lines. The significance of this study was to identify the optimal concentration of MS medium with growth hormones for improving the regeneration of Glycine max to reduce the period of regeneration using cotyledonary node.Transgenic crops can be developed which can be used not only for expression of different biochemical functions, but also for stress signal transduction that increase stress effects in plants. For cell protection, higher amount of proline and protein play an important role against oxidative stresses. Collectively, all these mechanisms allow plants to stay alive under extreme salt stress thereby leading to higher yield production of Glycine max.