In vitro plant regeneration was achieved from shoot explants of Amsonia orientalis Decne. The seeds were germinated aseptically in Petri dishes containing growth-regulator-free MS medium. The shoot explants of 30-day-old grown seedlings were cultured on MS medium supplemented with 1 mg/l indole-3-butryric acid (IBA) plus 0.5 mg/l kinetin (Kin) for direct regeneration and subcultured at three week intervals. While induction of adventitious buds from shoot explants was achieved approximately 33% within four weeks, a few roots (around 15%) were induced on the same medium within eight weeks. In addition, shoot explants were transferred to MS medium containing 2 mg/l benzyl aminopurine (BAP) plus 0.2 mg/l naphtalene acetic acid (NAA) for indirect regeneration and after three weeks, proliferated actively growing calli was occured.