Establishment of a competent protocol for high frequency plant regeneration is done by analyzing a range of factors such as genotypes, combinations of plant growth regulator, types of explants, age of explants etc. in present study maximum shoot regeneration frequency observed was 56.5% for cotyledonary petiole on Murashige and Skoog (MS) medium supplemented with 2 mg/L BA (6-benzyladenine) and 0.2 mg/L NAA (1-naphthalene acetic acid). Out of all the explants reported from Brassica species cotyledonary explants generated the highest frequency in regeneration of shoots. The regenerated plantlets were acclimatized, hardened and transferred to field and grown to maturity in the greenhouse. All the regenerated plants obtained were fertile and morphologically indistinguishable with the source plants.