Production of podophyllotoxin, kaempferol and quercetin in callogenesis of Dysosma versipellis — an endangered medicinal plant

Abstract

• Dysosma versipellis is an endangered medicinal herb found in southwest regions of China. • Callus initiation was achieved in B5 salts with 2 mg/L 2,4-D, and 0.2 mg/L kinetin under complete darkness. • The maximum callus biomass proliferation was obtained on B5 basal salts supplemented with 2 mg/L 2,4-D. • Leaf callus showed maximum production of podophyllotoxin, kaempferol and quercetin than that of control. • UPLC analysis revealed that roots and rhizomes were the primary source of secondary compounds accumulation. Dysosma versipellis is an endangered medicinal herb found in southwest regions of China. It has long been used in Traditional Chinese Medicine for general remedies in China. This herb is commonly used as an alternative to Podophyllum species. The large-scale removal of D. versipellis rhizomes from natural habitat continues at higher rates than natural regeneration. This study reports the callus induction from D. versipellis explants, callus proliferation and production of tumor inhibitory compounds such as podophyllotoxin, kaempferol and quercetin. The results revealed that tender leaves of D. versipellis exhibited the highest callus induction efficiency of 86% than other explants tested. The best callus initiation was obtained in Gamborg's medium (B5) augmented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 0.2 mg/L kinetin, 3% sucrose, 0.1% activated charcoal and 0.7% agar and incubated under complete darkness. The maximum callus biomass proliferation was obtained on B5 basal salts supplemented with 2 mg/L 2, 4-D. The UPLC analysis of leaf callus showed maximum production of podophyllotoxin, kaempferol and quercetin than that of control callus. Furthermore, analysis of whole plant parts for their secondary compounds found roots and rhizomes were the primary source of secondary compounds accumulation. Therefore, this bioactive-enriched callus biomass protocol may be used as a possible technique for sustainable production of podophyllotoxin, kaempferol and quercetin at commercial scale.