Optimization of Artemisinin Accumulation in Artemisia annua L. Green Callus Cultures: Impact of Media, Elicitors and Environmental Factors

Abstract

Introduction: Artemisinin is a sesquiterpene lactone with an endoperoxide bridge isolated from Artemisia annua L. It is effective against chloroquine-sensitive and chloroquine-resistant Plasmodium falciparum malaria parasites and is used in the treatment of drug-resistant malaria. Aim: This study used callus culture techniques to evaluate artemisinin accumulation in green callus tissue. Results: Artemisia annua leaves were used to regenerate green callus tissue. The callus-induction culture medium was LV medium supplemented with BA (1.0 mg/L), NAA (1.0 mg/L), sucrose (30 g/L), and a light intensity of 22.2 µmol/m²/s, resulting in 100% callus formation after 12 days. LV medium supplemented with casein hydrolysate (0.5 g/L) increased cell biomass to 19.963 g/cluster. The elicitors yeast extract and chitosan affected the artemisinin-accumulation capacity of green callus tissue. The optimal elicitor concentrations were 100 mg/L yeast extract and 100 mg/L chitosan, resulting in artemisinin accumulation of 0.0462% of dry cell weight when supplemented on day 14. Sucrose 30 g/L, casein hydrolysate/yeast extract 200/40 mg/L, KH₂PO₄ 200 mg/L, and light intensity of 22.2 µmol/m²/s increased artemisinin accumulation to 0.0735% of dry cell weight. The F10 generation showed the highest artemisinin accumulation, with a content of 0.0762%. Conclusion: LV medium, casein hydrolysate, yeast extract, sucrose, and light intensity influenced artemisinin accumulation in A. annua green callus tissue.