Copper sulphate elicitation of optimized suspension culture of andrographis paniculata nees yields unprecedented level of andrographolide

Abstract

Concerted efforts have been made to push up andrographolide production through optimization of culture conditions, calli induction, cell growth and elicitation in cell suspension culture of Andrographis paniculata Nees. Optimum callus induction was obtained with cotyledon and hypocotyls of the plant on Skoog and Hilderbrandt (SH) medium containing 2.0 μg/mL 2,4-dichlorophenoxyacetic acid and 0.1μg/mL 6-benzyl amino purine (BAP). Half MS medium containing 20 g/L sucrose and 20 h photoperiod showed highest cells fresh weight (CFW) (17.96 ± 0.06 g/50 mL), growth index (10.95 ± 0.96) and andrographolide yield (4.61±0.688 mg/g DCW). The addition of copper sulphate (500 μM/L), Methyl jasmonate (25 mg/L), Chitin (500 mg/L) or Fungal mycelium (500 mg/L) in separate experiments showed significant increase (p=0.05) in bioproduction of andrographolide to the extent of 29.42±0.31 mg/g DCW, 13.13±0.11mg/g DCW, 19.45±0.68 mg/g DCW and 13.629±1.12 mg/g DCW respectively, copper sulphate thus proved to be the most effective one. The study indicated that a holistic approach involving both culture conditions and elicitation could enhance the overall production of secondary metabolites (andrographolide) appreciably in this herb and possibly in other medicinal plants.