The present study aimed to investigate the effect of different plant growth regulator namely 2,4 dichlorophenoxy acetic acid (2,4-D) and ?-Naphthalene Acetic Acid (NAA) on callus induction of Helianthus annuus L. Callus culture were initiated from three explants namely cotyledon leaf, hypocotyl and root segments explants of in vitro Helianthus annuus L seedling. Both auxins 2,4-D and NAA stimulate callus induction, however 1.5 mg/L 2,4-D proved to be more effected for induction of callus. Among all treatment media containing 1.5 mg/l 2,4 D showed highest response of callus degree (3.44