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Embryogenic suspensions of adult cork oak: the first step towards mass propagation

Abstract

Protocols have been established to clone adult cork oak trees by somatic embryogenesis using semisolid medium. However, for economically viable mass propagation, embryogenic cultures in liquid medium need to be developed. In this study, suspension cultures were initiated from embryo clusters obtained by secondary embryogenesis on a gelled medium lacking plant growth regulators. After 6 days of culture, these embryo clusters generated high cell density suspensions that also contained small organized structures (embryos and embryogenic clumps). As the culture duration increased, tissue necrosis and fewer embryogenic structures were observed and the establishment of suspension cultures failed. An alternative method was found adequate for initiation of embryogenic suspensions: embryo clusters from gelled medium were briefly shaken in liquid medium and detached cells and embryogenic masses of 41–800 μm were used as inoculum. Maintenance of embryogenic suspensions was achieved using a low-density inoculum (43 mg l−1) by subculturing four embryogenic clumps of 0.8–1.2 mm per 70 ml of medium. Proliferation ability was maintained for almost 1 year through ten consecutive subcultures. The initiation and maintenance protocols first developed for a single genotype were effective when tested on 11 cork oak genotypes.

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Acknowledgments

The improvements of the manuscript by the anonymous reviewers and the communicating editor are highly appreciated. This work was funded by the Spanish National R&D Programme project AGL2007-66345-CO02-01. It was also supported by a postdoctoral fellowship from Instituto Nacional de Investigaciones Agrarias (INIA) to D. López-Vela and a postgraduate grant from Instituto Madrileño de Investigación y Desarrollo Rural, Agrario y Alimentario (IMIDRA) to J. Jiménez. This work is part of the requirements to fulfil the J. Jiménez’s Ph.D. degree.

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Correspondence to Jesús Alegre.

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Communicated by K. Klimaszewska.

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Jiménez, J., López-Vela, D., Ruiz-Galea, M. et al. Embryogenic suspensions of adult cork oak: the first step towards mass propagation. Trees 27, 13–23 (2013). https://doi.org/10.1007/s00468-012-0763-y

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  • DOI: https://doi.org/10.1007/s00468-012-0763-y

Keywords

  • Micropropagation
  • Quercus suber
  • Somatic embryogenesis
  • Suspension culture