Pigeonpea (Cajanus cajan) is ranked second after chickpea in India in area and production. It is predominantly consumed as pulse. Pigeonpea is a high protein grain legume, having less amount of sulfur containing amino acids like methionine, cystine along with tryptophan as compared to other legumes. These problems can be solved by plant tissue culture technique which has been used extensively as an alternate tool to accelerate the crop improvement process. In the present study the search for optimal culture conditions promoting adventitious shoot formation and plantlet regeneration in pigeonpea has been done. This study showed that Bahar responded better in comparison to other varieties in Blayde’s medium for regeneration purpose. It encouraged us to standardize the regeneration protocol of Bahar variety from leaf and stem explants along with mature and decapitated embryos. Among these leaf explants and decapitated embryo proved better than stem explants for regeneration purposes. The isoenzyme patterns of above varieties explants have also been studied.