In the present investigation, a simple, rapid and efficient in vitro regeneration system was developed in green gram (Vigna radiata L. Wilczek) cv. IPM-02-03 using cotyledonary explants excised from a 4 day old seedling. Organogenic calli were developed from cotyledonary tissues within 4-6 weeks of culture, supplemented with 2.5 mg/L BAP and 1.5 mg/L NAA. Shoot regeneration was achieved from shoot tips and cotyledonary node on MS medium was supplemented with BAP 2.0 mg/l. Regeneration frequency depends on genotype, explants types and growth regulator combinations in the medium. 1.5 mg/l BAP in combination with 1.0 mg/l Kinetin was proved to be the most effective treatment for multiple shoots induction from nodal explants after 3-4 weeks. The number of shoots per culture varied from 1.3 to 7.8 in different growth media. The cultures were incubated for 16 hrs of photoperiod in 25^oC for 2 weeks for shoot bud regeneration. The proliferated shoots were further sub cultured on similar medium for higher shoot bud regeneration. Rooting was achieved in all rooting media but elongated shoots were rooted on ½ strength MS media supplemented with 0.5 mg/l NAA or IBA with 2% (W/V) sucrose which produced the maximum number of strong and healthy roots. The rooted plantlets were transferred to soil mixture (Soil: Sand: Vermicompost 1:1:1 ratio) and kept in the green house with 85% humidity. Seventy-five percent plants were successfully grown after regeneration with no phenotypic variation. This protocol can be used for genetic transformation and large-scale production of genetically improved green gram variety.

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