In vitro cultures of Berberis buxifolia were established using thidiazuron (4.5, 23 and 45 mM) or picloram (4 and 40 mM) as plant growth regulators for sustaining growth. For producing berberine, a two-stage culture was performed. In the first step, thidiazuron or picloram were used for biomass production followed by the production stage where benzylaminopurine (4.4 mM) was added as a plant growth regulator. Berberine yields (102 mg g(-1) DW) and in vitro shoot cultures (200 mg g(-1) DW) were significantly lower than those of whole plants in the field (416 mg g(-1) DW). The highest productivity (0.18 mg 1(-1) day(-1)) was attained using picloram (either 4 on 40 mM) in the first stage for producing biomass.