Micropropagation and greenhouse cultivation of Scrophularia takesimensis Nakai, a rare endemic medicinal plant

摘要

A protocol was developed for direct shoot regeneration of Scrophularia takesimensis using shoot tip explants. Theexplants were excised from mature field grown plants and cultured on Murashige and Skoog (MS) medium with differentconcentrations of 2 isopentyl adenine (2 iP), 6 benzyl adenine (BA) or thidiazuron (TDZ). Among the three cytokininsstudied, BA was found to be the most effective cytokinin for multiple shoot induction. Addition of auxin to the shootregeneration medium significantly enhanced the percentage of shoot induction and number of shoots per explant. Thegreatest percentage of shoot induction was achieved when shoot tip explants were cultured on MS medium supplementedwith 3.0 mg L1BA and 1.0 mg L1indole 3 acetic acid (IAA) with an average of 13 shoots per explant. The regeneratedshoots were transferred to modified MS medium supplemented with 3% (w/v) sucrose, 1.0 mg L1indole 3 butyric acid(IBA), and solidified with 0.8% (w/v) agar for four weeks to induce the growth of shoots and roots. P lantlets weretransferred to the greenhouse with 100% survival rate. High performance liquid chromatography analysis detected thepresence of harpagoside in greenhouse grown plants organs which were established from in vitro culture. The content ofharpagoside was high in fruit followed by leaf, root and stem. This protocol could be utilized for conservation and clonalpropagation and chemical analysis of this economically important plant.