Abstract: Three different explants; leaf, root and stem were taken from in vitro grown seedlings to observe shoot regeneration, callus induction and rooting Sunflower (Helianthus annuus L.). Shoots were regenerated on MS basal medium containing 0.1mg/l α-naphthalene acetic acid (NAA) with six different concentration of 6-benzylaminopurine (BAP) (0.0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0mg/l). The regenerated shoots were induced root on MS basal medium with Indole-3- butyric acid (IBA) (0.01, 0.1, 1.0 mg/l). Callus was induced on MS basal medium with 0.1 mg/l BAP + 2, 4 D (0.0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0mg/l). Completely Randomized Design (CRD) with 10 replicates was used. Data was analyzed using SAS program (9.1.3).