Establishment of cell suspension cultures of yew (taxus××media rehd.) and assessment of their genomic stability
2005
BAEBLER, ŠPELA | HREN, MATJAŽ | CAMLOH, MARJANA | RAVNIKAR, MAJA | Bohanec, Borut | PLAPER, IGOR | UCMAN, ROBERT | Žel, Jana
Yew cell suspension culture is used as an alternative source of paclitaxel, an anti-cancer drug. To optimize the initiation protocol, highly dormant yew seeds were germinated in vitro and the seedlings used to establish callus culture. The best sources of explant for callus initiation and growth were seedling stems and roots, and the most successful medium was modified B5 medium containing 2,4-dichlorophenoxyacetic acid and kinetin. Calluses were friable and suitable for establishing cell suspension cultures, which were maintained for over 3 yr. Flow cytometric analysis of nuclear DNA content revealed that 2-yr-old cell suspension cultures consisted predominantly of putative euploid and aneuploid cells coexisting as sub-populations. Additional measurements performed 3 and 7 mo. later revealed further genomic instability, with a tendency towards a higher proportion of cells with elevated nuclear DNA content. In a selected cell line, which showed significant taxane production, the addition of 100 μμM jasmonic acid strongly enhanced total taxane production and slightly inhibited growth while no effect on nuclear DNA content was noted.
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