Sorghum is one of the world's fifth most important grain crops, staple food for many people in Africa and Asia and a major feed crop for domesticated animals. In the present study, we investigated on three areas of aspects in Sorghum genotypes i.e., Tissue culture, estimation of bioactive compounds with antioxidant activity and finally seed proteomic analysis. An attempt has been made in this study to establish a plant tissue culture protocol for efficient callus induction and subsequent plant regeneration system in Sorghum. The present study has achieved efficient callus induction (100%), enhanced regeneration from different explants of Sorghum genotypes IS 3477 (non pigmented), . Where as antioxidant activity has shown pigmented line IS 2898, finally total proteome map has shown in IS 3477, IS 33095 pigmented genotypes. The conclusion of this study when compared pigmented and non pigmented sorghum genotypes to identification of differential genotypic variation for the plant breeders to improve their yield crop.