Chitosan- and salicylic acid-mediated enhancement of andrographolide in Andrographis paniculata (Burm.f.) Wall. leaf callus

Andrographis paniculata (Burm.f.) Wall., widely known as the “King of Bitters,” is valued for its medicinal diterpenoid andrographolide. The present study investigated the effects of surface-sterilization treatments, plant-growth regulators (PGRs), and elicitors on callus induction and andrographolide accumulation in A. paniculata leaf explants. Leaf tissues sterilized with 60 % sodium hypochlorite for 1 min achieved 83.33 % survival and 96.67 % aseptic culture. The optimal callus formation occurred on MS medium containing 1.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) + 0.5 mg L−1 kinetin, giving 96 % callus induction within 8.4 ± 0.0 days, 2.8 ± 0.0 g fresh weight, and 2.2 ± 0.0 cm diameter. Chitosan at 5 mg L−1 produced 100 % survival and markedly enhanced biomass (3.0 ± 0.0 g; 2.5 ± 0.0 cm diameter). High-performance liquid chromatography (HPLC) analysis revealed that 5 mg L−1 chitosan increased andrographolide content to 962.07 ± 0.27 μg g−1 approximately 6.8-fold higher than the control (123.44 ± 0.16 μg g−1). These results establish a reproducible protocol for callogenesis of A. paniculata and demonstrate that elicitation substantially improves callus growth and andrographolide yield, offering a sustainable platform for secondary-metabolite production.

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