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Establishment of cell suspension cultures of yew (Taxus × media Rehd. and assessment of their genomic stability

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Summary

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 μM jasmonic acid strongly enhanced total taxane production and slightly inibited growth while no effect on nuclear DNA content was noted.

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Authors and Affiliations

  1. Department of Plant Physiology and Biotechnology, National Institute of Biology, Večna pot 111, 1000, Ljubljana, Slovenia

    Špela Baebler, Matjaž Hren, Marjana Camloh, Maja Ravnikar & Jana Žel

  2. Biotechnical Faculty, University of Ljubljana, Jannikarjeva 101, 1000, Ljubljana, Slovenia

    Borut Bohanec

  3. Krka d.d., Šmarješka cesta 6, 8000, Novo mesto, Slovenia

    Igor Plaper & Robert Ucman

Authors
  1. Špela Baebler
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  2. Matjaž Hren
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  3. Marjana Camloh
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  4. Maja Ravnikar
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  5. Borut Bohanec
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  6. Igor Plaper
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  7. Robert Ucman
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  8. Jana Žel
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Correspondence to Špela Baebler.

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Baebler, Š., Hren, M., Camloh, M. et al. Establishment of cell suspension cultures of yew (Taxus × media Rehd. and assessment of their genomic stability. In Vitro Cell.Dev.Biol.-Plant 41, 338–343 (2005). https://doi.org/10.1079/IVP2005642

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  • DOI: https://doi.org/10.1079/IVP2005642

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