Plumbago zeylanica L. is a traditional herbal that has been reported to treat on skin diseases. Furthermore, some researchers have found plumbagin extracted from roots of this species can prevent cancer cell development. In current study, stems of Plumbago zeylanica L. were cultured on MS medium with BA 1.0 mg/L and IAA (0.01-0.15 mg/L) or NAA (0.1-0.15 mg/L). After 8-week cultured, stems were transferred to MS medium with extracted from stems of Portulaca grandiflora Hook (2-10 ml/L) or extracted from stems of Ipomoea batatas (L.) Poir. The results showed that, the appropriate medium for shoot formation was in MS with BA and IAA 0.1 mg/L or NAA 0.1 mg/L. The adventitious roots in vitro were formatted in MS medium supplied with extracted from stems Portulaca grandiflora Hook or from stems of Ipomoea batatas (L.) Poir 6 ml/L. Simultaneously, after 8-week cultured, the adventitious roots were collected and plumbagin qualitative were analyzed with pure plumbagin of Sigma. As the results, plumbagin presents in adventitious roots cultured.