Optimization of Artemisinin Accumulation in Artemisia annua L. Green Callus Cultures: Impact of Media, Elicitors and Environmental Factors
Mai Thi Phuong Hoa
Nguyên Tat Thanh University, HCM, Vietnam.
Đo Tien Vinh
Nguyên Tat Thanh University, HCM, Vietnam.
Pham Hong Diep
International University, Vietnam National University, HCM, Vietnam.
Tran Van Minh *
International University, Vietnam National University, HCM, Vietnam.
*Author to whom correspondence should be addressed.
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.
Keywords: Artemisia annua L., artemisinin, callus culture technique, green callus tissue, elicitor, yeast extract, chitosan, casein hydrolysate, sucrose, light intensity, secondary metabolites, plant tissue culture