Abstract

Artemisia monosperma (Delile) callus was induced using seedling explant cultured on Murashige and Skoog solid medium (M&S) supplemented with 1 mg/L naphthalene acetic acid (NAA) and 1 mg/L kinetin (Kn). Cultures were maintained on 2 mg/L 2,4-dichlorophenoxy acetic acid (2, 4- D) and 1 mg/L Kn and treated with different elicitors. Total flavonoids and phenolics were determined by aluminum chloride-potassium acetate and Folin-Ciocalteu colorimetric methods, respectively. Yeast extract 10 mg/L (Y2) showed higher productivity and viability of callus cultures than Fusarium oxysporum and calcium chloride elicitors. Y2 shows an increase of 2.4 and 1.5 times non-elicited calli for flavonoids and phenolic compounds production, respectively. Pockets of embryogenic calli were transferred to M&S solid media supplemented with 0.5 mg/L NAA and 0.5 mg/L benzyl aminopurine (BAP), followed by hormonal free media where different stages of embryos were monitored, giving regenerated plantlets. HPLC analysis of methylene chloride and ethyl acetate fractions of parent plant (P), Y2 elicited embryogenic callus (EC.Y2) and Y2 elicited non-embryogenic callus (NEC.Y2) showed that, quercetin content in methylene chloride fraction of NEC.Y2 is 7.2 times the same fraction of P, while vanillic acid ethyl ester content in ethyl acetate fraction of EC.Y2 (EEC.Y2) is 8.6 times the same fraction of P. EEC.Y2 showed highest anti-oxidant activity with IC₅₀ 7.22±0.14 µg/mL compared with ethyl acetate fraction of P with IC₅₀ 20.01±0.82 µg/mL (IC₅₀ of L-ascorbic acid = 1.24±0.07 µg/mL). Using MTT assay, EEC.Y2 exhibited potent cytotoxic activity against colon carcinoma cells and moderate activity against hepatocellular and lung carcinoma cells with IC₅₀ 18.9±1.4, 22.3±0.9 and 41.6±1.2 µg/mL, respectively; compared with doxrubcin as reference standard.

Key words: Artemisia monosperma, callus cultures, flavonoids, phenolic compounds, anti-oxidant, cytotoxicity.