In vitro cell culture of Charybdis congesta for enhanced production of secondary metabolites: Proscillaridin A, Scillaren A and Scilliroside

Callus cultures of Charybdis congesta were initiated in vitro and the effect of growth regulators was tested on callus growth and secondary metabolite production. Among several standard media formulated for use in the present study, MS and B5 were found to be potentially active and facilitated the calculation of callus induction frequency (CIF). The CIF was higher in both MS (70%) and B5 (63%) media supplemented with 1-naphthalene acetic acid (NAA) (9.0 mu M) and benzyl ammo purine (BAP) (0.9 mu M). However, with indole-3-butyric acid (IBA) (9.0 mu M) and BAP (0.8 mu M), less amount (22.6%) of CIF was observed in MS medium but no callus formation was noticed in B5 medium. Rapid high performance thin layer chromatography (HPTLC) screening of callus extracts revealed that the callus established in MS medium supplemented with 4.5 mu M NAA and 0.46 mu M BAP produced the highest yield of Proscillaridin A (4.51 mg/g DW), Scilliroside (3.3 mg/g DW), Scillaren A (2.35 mg/g DW) and desacetylscilliroside (8.62 mg/g DW), which was higher than from the intact plants. The results obtained indicate that the in vitro cultures of C. congesta might be an excellent source of secondary metabolites and further metabolic profiling may provide insights into up scaling of the compounds which lead to greater commercial interest and continuous supply of cultures.