Abstarct: For optimization of tissue culture of Echinacea Purpurea medicinal plant, this study experiments was carried out as a factorial arrangement in a completely randomized design. In the first experiment, the effect of different concentrations of cytokinin hormones include 6-benzylaminopurine (BAP) and Thidiazuron (TDZ) and kinetin (KIN ( alone or in combination with different concentrations of auxin hormones include indole-3-butyric acid (IBA) and naphthaleneacetic acid (NAA) in callus induction in leaf and petiol explants evaluated on basal MS medium. The results showed that the highest percentage callogenesis (100%) obtained in combinations include TDZ and IBA hormones. In the second experiment, the effect of 2 mg/L BAP hormone in shoot organogenesis in callus of leaf and petiol explant evaluated. Higher rates (92/85%) of shoot organogenesis were observed on MS medium supplemented with NAA and BAP from leaf explant. In the third experiment, the effect of different concentrations 6benzyl amino purine (BAP) and Thidiazuron (TDZ) Hormones alone or in combination with different concentrations of naphthaleneacetic acid (NAA) in shoot organogenesis in leaf explant evaluated. Higher rates (85%) of shoot organogenesis were observed with 2 mg/l BAP & 0.1 mg/l NAA combination. In the fourth experiment plantlets from hormone combination included 2 mg/l BAP & 0.1 mg/l NAA transferred to MS medium supplemented with NAA and IBA. Highest number root per explant (8/16) was achieved using MS media supplemented with 0/5 mg/l NAA.