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Cloning mature holm oak trees by somatic embryogenesis

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Integuments from holm oak developing ovules were suitable initial explants to obtain embryogenic lines from which plants could be regenerated.

Abstract

The implementation of multivarietal forestry as part of breeding strategies is expected to provide more productive forest plantations. To achieve this, a reliable and effective method for mass production of clonal plants is needed. Somatic embryogenesis is considered the enabling technology for implementing multivarietal forestry. The holm oak (Quercus ilex L.) is a Mediterranean evergreen tree of economic interests because of the acorn production for animal feed and edible fungi mycorrhization. The aim of this work was to obtain clonal plants by inducing somatic embryogenesis in tissues of female flowers from mature trees. The influence of the developmental stage of the explant, the genotype and medium composition, and the effect of arabinogalactan proteins on the induction frequency, were assessed. Somatic embryogenesis induction (frequency ranging from 1.2 to 3.2 %) was restricted to ovules excised at an advanced stage of development, when they were at least 3–4 mm wide and the rest of the ovules within the ovary had aborted. Somatic embryos arose from the integuments of those fertilized ovules. Embryogenic response was obtained on media with and without plant growth regulators. All the genotypes that were cultured on medium containing “as reported by Schenk and Hildebrandt (Can J Bot 50:199–204, 1972)” SH macronutrients could be captured. Treatments including Larix arabinogalactan proteins did not improve induction, while those from Acacia inhibited the embryogenic response. Several embryogenic lines were multiplied by repetitive embryogenesis on medium lacking plant growth regulators. Mature somatic embryos of three genotypes were germinated at frequencies ranging from 41 to 58 %, and converted into plants at frequencies from 11 to 30 %, depending on the genotype.

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Acknowledgments

This work was funded by The Spanish Ministry of Science and Innovation (AGL2010-22292-C01 and C03 projects). It was also supported by a postgraduate fellowship from IMIDRA to A. Barra-Jiménez. This work is part of the requirements to fulfill the A. Barra-Jiménez’s Ph.D. degree.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Instituto Madrileño de Investigación y Desarrollo Rural, Agrario y Alimentario (IMIDRA), Finca El Encín, Apdo. 127, 28800, Alcalá de Henares, Madrid, Spain

    Azahara Barra-Jiménez, Mar Ruiz-Galea, Cristina Celestino, Jesús Alegre & Mariano Toribio

  2. ERI Biotecmed, Dpto. de Biología Vegetal, Universidad de Valencia, Avda. Vicent Andrés Estellés s/n, 46100, Burjassot, Valencia, Spain

    Miquel Blasco & Isabel Arrillaga

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  1. Azahara Barra-Jiménez
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  2. Miquel Blasco
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  3. Mar Ruiz-Galea
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  4. Cristina Celestino
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  5. Jesús Alegre
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  6. Isabel Arrillaga
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  7. Mariano Toribio
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Correspondence to Mariano Toribio.

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Communicated by K. Klimaszewska.

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Barra-Jiménez, A., Blasco, M., Ruiz-Galea, M. et al. Cloning mature holm oak trees by somatic embryogenesis. Trees 28, 657–667 (2014). https://doi.org/10.1007/s00468-014-0979-0

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