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Enhanced taxane production was observed in a hybrid, two liquid phases containing, cultures of Taxus x media var. Hicksii hairy root carrying a taxadiene synthase transgene, supported with liquid perfluorodecalin (PFD) in degassed or aerated form. The hairy root cultures were elicited with methyl jasmonate (MJ, 100 μM) or coronatine (COR, 1 μM), and fed with sucrose and l-phenylalanine. The root growth was not stimulated by PFD addition, irrespective of the day of its application (day 0 and 14). However, in the cultures elicited with MJ and performed in the presence of PFD the final root biomass accumulation was higher than in cultures performed without PFD while the opposite effect was observed in cultures supplemented with COR. The highest paclitaxel content in root biomass was determined at the end of the cultures elicited with MJ and supplemented with PFD-degassed at day 0 or 14, 1,440.8 and 1,432.5 μg g−1 DW, respectively. The highest total (i.e. intracellular + extracellular: both in aqueous and PFD phases) paclitaxel yield in flasks (149.15 μg flask−1) was noted after the application of PFD-degassed at day 14. The other taxane detected was baccatin III, only in the root biomass, with the highest content (76.9 μg g−1 DW) observed under COR treatment. Although COR stimulated paclitaxel production with less efficiency than MJ, it resulted in higher paclitaxel excretion to the liquid phases of culture medium and PFD.

plant cell, tissue and organ culture

Perfluorodecalin-supported system enhances taxane production in hairy root cultures of Taxus x media var. Hicksii carrying a taxadiene synthase transgene