Soybean is one of the most important legumes of the world and it is a rich source of dietary proteins and used as raw and processed food as well as in drink. Recent studies have shown that soybean may be helpful in preventing bone loss and may help to prevent or slow the development of breast cancer and also lower the risk of type 2 diabetes. The present study was conducted to analyse the effect of plant growth regulators on direct and indirect organogenesis of soybean (JS335). In direct regeneration of cotyledonary node explants, the highest percentage of regenerated shoots (80%) were observed in the 15 μM of TDZ with the maximum number of shoots (8.33±0.33). The 20 μM of IBA showed the best response of root formation with maximum number of roots (41.6±1.20) and length of roots (15.36±0.24 cm). Among the different concentrations of hormones tested for callus induction the best response was noticed in the MS medium supplemented with BAP from half seed explants. The highest frequency of regeneration (83.3%) was obtained when the calli were transferred into the shooting medium containing BAP (15 μM) with the maximum number of shoots per explant (8.66±0.88). Shoots regenerated through callus were rooted well in the concentration of 15 μM IBA. Then the rooted plantlets were transferred into the soil with paper cups.