초록
An in vitro culture of Artemisia absinthium L. was established for production of commercially important secondary metabolites. Callus cultures were obtained by inoculating leaf explants on Murashige and Skoog (MS) medium supplemented with Thidiazuron (TDZ; 0.5-5.0mgl<SUP>-1</SUP>) alone or in combination with either α-naphthalene acetic acid (NAA; 1.0mgl<SUP>-1</SUP>) or Indole acetic acid (IAA; 1.0mgl<SUP>-1</SUP>). The callus obtained in response to 1.0mgl<SUP>-1</SUP> TDZ and 1.0mgl<SUP>-1</SUP> NAA was subcultured on the same medium to investigate its biomass accumulation and secondary metabolites production on weekly basis for 7 weeks. For submerged cultivation, 35 day old calli were cultured on MS basal media supplemented with 1.0mgl<SUP>-1</SUP> TDZ and 1.0mgl<SUP>-1</SUP> NAA. Growth kinetics and secondary metabolites production were investigated in 3 day old suspension cultures for 42 days. Additionally, high performance liquid chromatography (HPLC) based quantification of gallic acid, caffeic acid and catechin was carried out in cell suspension cultures. Seed germinated plantlets were used as control. Maximum levels of total phenolic content 3.57mgGAE/gDW (control: 2.75mgGAE/gDW), total flavonoid content 1.89mgQE/gDW (control: 1.20mgQE/gDW), and antioxidant activity 82.7% (control: 72.3%) were displayed by suspension cultures. Among the phenolic compounds, maximum level of gallic acid 104μgg<SUP>-1</SUP> (control: 21.3μgg<SUP>-1</SUP>), caffeic acid 27.40μgg<SUP>-1</SUP> (control: 28.5μgg<SUP>-1</SUP>) and catechin 92.0μgg<SUP>-1</SUP> (control: 68.10μgg<SUP>-1</SUP>) were detected in suspension cultures. The results indicate that cell suspension cultures of A. absinthium L. have the potential for enhanced production of phenolics and, hence, highest antioxidant activity than callus culture and seed derived plantlets.