Root cultures of Duboisia myoporoides and D. leichhardtii have been established from granular tissues isolated from a culture line of callus. The granular tissues easily differentiated roots which vigorously grew in liquid Murashige-Skoog medium supplemented with indole-3-butyric acid (IBA) (2 mg/l) and gibberellic acid (1 mg/l).These cultured roots produced atropine, scopolamine and nicotine. Furthermore, anabasine and nornicotine were detected in root cultures of D. myoporoides, and apoatropine in D. leichhardtii root cultures. A high concentration (7-10% (w/v)) of sucrose in the medium was effective in improving both root growth and tropane alkaloid production.