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.
Highlights
Secondary metabolism in plants gives rise to a wide range of chemical compounds including coumarins, alkaloids, phenolics and terpenoids that may provide defense against pathogenic infections (Harborn,1982)
Among different combinations of auxins and cytokinins that were tested with B5 and Skoog and Hilderbrandt (SH) media, the highest callus induction (93.33 ± 1.66%) was shown by cotyledon explants on SH medium at 2, 4-D (2.0 μg/mL) and benzyl amino purine (BAP) (0.1 μg/mL) followed by leaflet explants (83.33 ± 2.89 %) on SH at 2, 4-D (1.0 μg/mL)
In both SH and B5 media, 2, 4-D alone produced compact callus from cotyledons but when 2, 4-D was applied with BAP and KIN friable calli were obtained in most cases
Summary
Secondary metabolism in plants gives rise to a wide range of chemical compounds including coumarins, alkaloids, phenolics and terpenoids that may provide defense against pathogenic infections (Harborn,1982). These secondary metabolites form the basis for the development of new pharmaceutical drugs, herbicides, cosmetics and pesticides (Rodriguez-Concepscion and Boronat, 2002). Andrographolide is a potential medicine to control asthma and other inflammatory diseases via inhibition of the Nuclear Factor-κB Pathway (Bao et al, 2009) and may be applied in the treatment of cancer, autoimmune diseases (Hahn and Albersheim, 1978) and idiopathic pulmonary fibrosis (Zhu et al, 2013). Andrographolide nanoparticle has been found to possess hepato-protective, anti- bacterial anti-malarial (anti-plasmodial) and anti-viral (Harborn, 1982; Valiani et al, 2011) activities and shown to protect against cigarette smoking induced chronic obstructive pulmonary disease (COPD) (Guan et al, 2013)
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