Angelika FILOVÁ – Eleonóra KRIVOSUDSKÁ
Abstract: In this study, callus cultures from leaves and young shoots of Taxus baccata L.were established in Gamborg’s B5 medium supplemented with 2,4-dichlorophenoxiacetic acid (2 mg/L), kinetin (0.5 mg/L) and gibberellic acid (0.25 mg/L). Callus growth and taxane production were evaluated using two culture media: Woody Plant Medium and Gamborg’s B5 supplemented with picloram (2 mg/L), kinetin (0.1 mg/L) and gibberellic acid (0.5 mg/L). The effect of the inoculum size (50, 100 and 150 g FW/L) and culture media (Woody Plant Medium and Gamborg’s B5) with and without the presence of methyl jasmonate (100 M) on T. baccata cell suspensions was assessed. Taxane analysis revealed that the calus in Gamborg’s B5 produced taxol (50 g/g DW), baccatin III, 10-deacetyl baccatin III and 10-deacetyl taxol. Woody Plant Medium also induced the production of taxol, although to a lesser extent. The optimum inoculum size was 50 g FW/L. In cell suspension cultures, both media had a significant effect on taxane production when supplemented with methyl jasmonate. In WPM, at day 14, a total concentration of 185.35 g/L of taxol, 172.98 g/L of baccatin III, 658.97 g/L of 10-deacetyl baccatin III and 259.75 g/L 10-deacetyl taxol were obtained, with total excretion of baccatin III and 10-deacetyl taxol to the culture medium. The best culture conditions for producing taxol were found to be WPM supplemented with MeJ. The taxol level achieved in these conditions was 3.4 higher than in the same medium without elicitation and over 9 times higher than in the cultures grown in B5, elicited or not.
anticancer agent, in vitro cultures, methyl jasmonate, taxanes, Taxus baccata L.
Presentation: oral