Establishment of callus and cell suspension cultures of Granny Smith apple fruit and antityrosinase activity of their extracts

The purpose of the present study was to induce callus and suspension cell cultures from the fruit of Granny Smith apple to measure some of the phenolic compounds in cell culture and evaluate the skin anti-aging effect through measuring the anti-tyrosinase activity. For callus induction MS medium supplemented with different concentrations of plant growth regulators, 2,4-D + BA and 2,4-D + Kin were used. MS medium with 2 mg/l 2,4-D + 1 mg/l BA produced maximum callus formation of 100% plus an average fresh weight and dry weight of 2.754 gr and 0.37 g, respectively. In addition, this combination of growth regulators induced callus in the shortest time (6.6 days). It was also found that the best callus quality, friable and white-yellowish, was achieved on MS medium containing 2,4-D + BA. The cell suspension cultures from callus with 2,4-D + BA were established properly and the maximum of fresh and dry biomass growth (4.53 and 0.65 g L-1) was obtained in 20 days of culture. Also, cell viability and distribution as singular or aggregated masses were evaluated using fluorescent microscopy after staining with the FDA. HPLC-based quantification of chlorogenic acid, caffeic acid, catechin, and quercetin was carried out on cell culture extracts. The result showed methanolic extract contained 1.42 mu g/mg chlorogenic acid, 0.46 mu g/mg catechin, 0.12 mu g/mg caffeic acid and 0.95 mu g/mg quercetin. The anti-tyrosinase activity of cell culture extract was measured and the highest inhibitory activity was related to 100 mg/mL extract (55%). IC50 was 1.21 mg/mL compared to that of Kojic acid as a positive control that was 0.0177 mg/mL.