Production of lithospermic acid B and rosmarinic acid in callus tissue and regenerated plantlets of salvia miltiorrhiza

Callus tissue from petioles of Salvia miltiorrhiza was obtained on Murashige and Skoog medium supplemented with indole-3-butyric acid and 6-benzylaminopurine. When the calli were subcultured, adventitious shoots formed. These shoots developed into normal plantlets with roots when transferred to hormone-free Murashige and Skoog medium. Clonal micropropagation was established by shoot tip culture and was maintained on Murashige and Skoog medium supplemented with kinetin and gibberellic acid A₃. Callus that was induced and maintained on supplements of 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine in the absence of light produced rosmarinic acid (1.24% dry wt) and lithospermic acid B (0.10% dry wt). Higher concentration levels of rosmarinic acid (6.96% dry wt) and lithospermic acid B (6.05% dry wt) were formed in the leaves of 15-week-old plants that were generated by micropropagation.