Establishing in vitro Zinnia elegans cell suspension culture with high tracheary element differentiation

Authors

  • Peter Twumasi,

    Corresponding author
    1. Laboratory of Plant Cell Biology, Department of Plant Sciences, Wageningen University and Research centre, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands
    2. Horticultural Production Chains Group, Department of Plant Sciences, Wageningen University and Research Centre, Marijkeweg 22, 6709 PG Wageningen, The Netherlands
    3. Department of Biochemistry and Biotechnology, Kwame Nkrumah University of Science and Technology (KNUST), Kumasi, Ghana
      Department of Biochemistry and Biotechnology, Kwame Nkrumah University of Science and Technology (KNUST), PMB, Kumasi, Ghana. Tel.: +233 245 131806; fax: +233 516 4338. E-mail addresses: twumasipeter@gmail.com
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  • Jan H.N. Schel,

    1. Laboratory of Plant Cell Biology, Department of Plant Sciences, Wageningen University and Research centre, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands
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  • Wim Van Ieperen,

    1. Horticultural Production Chains Group, Department of Plant Sciences, Wageningen University and Research Centre, Marijkeweg 22, 6709 PG Wageningen, The Netherlands
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  • Ernst Woltering,

    1. Agrotechnology and Food Innovation, Wageningen University and Research Centre, Wageningen, The Netherlands
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  • Olaf Van Kooten,

    1. Horticultural Production Chains Group, Department of Plant Sciences, Wageningen University and Research Centre, Marijkeweg 22, 6709 PG Wageningen, The Netherlands
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  • Anne Mie C. Emons

    1. Laboratory of Plant Cell Biology, Department of Plant Sciences, Wageningen University and Research centre, Arboretumlaan 4, 6703 BD Wageningen, The Netherlands
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Department of Biochemistry and Biotechnology, Kwame Nkrumah University of Science and Technology (KNUST), PMB, Kumasi, Ghana. Tel.: +233 245 131806; fax: +233 516 4338. E-mail addresses: twumasipeter@gmail.com

Abstract

The Zinnia elegans mesophyll cell culture is a useful system for xylogenesis studies. The system is associated with highly synchronous tracheary element (TE) differentiation, making it more suitable for molecular studies requiring larger amounts of molecular isolates, such as mRNA and proteins and for studying cellulose synthesis. There is, however, the problem of non-uniformity and significant variations in the yields of TEs (%TE). One possible cause for this variability in the%TE could be the lack of a standardized experimental protocol in various research laboratories for establishing the Zinnia culture. Mesophyll cells isolated from the first true leaves of Z. elegans var Envy seedlings of approximately 14 days old were cultured in vitro and differentiated into TEs. The xylogenic culture medium was supplied with 1 mg/l each of benzylaminopurine (BA) and α-naphthalene acetic acid (NAA). Application of this improved culture method resulted in stable and reproducible amounts of TE as high as 76% in the Zinnia culture. The increase was mainly due to conditioning of the mesophyll cell culture and adjustments of the phytohormonal balance in the cultures. Also, certain biochemical and cytological methods have been shown to reliably monitor progress of TE differentiation. We conclude that, with the adoption of current improvement in the xylogenic Z. elegans culture, higher amounts of tracheary elements can be produced. This successful outcome raises the potential of the Zinnia system as a suitable model for cellulose and xylogenesis research.

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