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Taxus globosa is the only species of the Taxus genus that grows in Mexico. In this study, callus cultures from leaves and young shoots of T. globosa 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 μM) on T. globosa cell suspensions was assessed. Taxane analysis revealed that the calli in Gamborg’s B5 produced taxol (50 μg/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 Woody Plant Medium, at day 14, a total concentration of 197.999 μg/L of taxol, 160.622 μg/L of baccatin III, 633.724 μg/L of 10-deacetyl baccatin III and 229.611 μg/L 10-deacetyl taxol were obtained, with total excretion of baccatin III and 10-deacetyl taxol to the culture medium. In Gamborg’s B5, cephalomanine was obtained at a concentration of 91.428 μg/L without elicitation, and all taxanes were excreted to the medium to a variable extent.
Taxus globosa Schltdl, the Mexican yew, represents a new source of taxanes, including taxol, baccatin III, 10-deacetylbaccatin III, 10-deacetyltaxol and cephalomannine. Due to the anticancer activity and other biological activities of these compounds, and their scarcity in nature, we initiated in vitro cultures of this species with the aim of developing a biotechnological process for obtaining taxol and related taxanes. In the current work, in a batch-type twophase culture of T. globosa, we evaluated the effect of cell immobilization and methyl jasmonate (MeJ) elicitation in two culture media containing different plant growth regulator combinations: 2,4-dichlorophenoxyacetic + benzylaminopurine (Treatment 1: T1) and picloram + kinetin (Treatment 2: T2). The productivity and excretion rate into the culture medium of baccatin III (12.79 lg L-1 d-1) (84 %), 10-deacetylbaccatin III (15 lg L-1 d-1) (0%), 10-deacetyltaxol (3.18 lg L-1 d-1) (63 %), and cephalomannine (49.27 lg L-1 d-1) (9 %) were increased by the effect of T1 in the free cell cultures elicited with MeJ. Cell immobilization in alginate beads did not improve the biotechnological production of these four taxanes. In contrast, the highest productivity of taxol (53 lg L-1 d-1) was achieved in MeJ-elicited free cells under T2 and cell immobilization in these conditions increased productivity by more than twofold (130.35 lg L-1 d-1).
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