Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 131, Issue 2, pp 321–333 | Cite as

Propagation of mature Quercus ilex L. (holm oak) trees by somatic embryogenesis

  • M. T. Martínez
  • M. C. San José
  • A. M. Vieitez
  • M. J. Cernadas
  • A. Ballester
  • E. Corredoira
Original Article
First Online:
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Accepted:

Abstract

Somatic embryogenesis from in vitro leaf and shoot apex explants excised from axillary shoot cultures established from two mature Quercus ilex trees has been developed. Somatic embryos (SE) were obtained from both explant types and genotypes evaluated, although embryogenic frequencies were influenced by the genotype, auxin concentration, and explant type. The explants were cultured on Murashige and Skoog salts and vitamins, supplemented with 500 mg L−1 casein hydrolysate (CH) and different concentrations of indole-3-acetic acid or α-naphthalene acetic acid (NAA) in combination with 2.22 µM 6-benzylaminopurine (BA). In both genotypes, shoot apex explants were more responsive than leaf explants. The best results were obtained with apex explants of clone Q3 (11%) cultured on medium with 21.48 µM NAA plus 2.22 µM BA. This combination was also effective for initiating SE from leaf explants, although the induction rates were lower (1–3%). Embryogenic lines were maintained by repetitive embryogenesis following culture of nodular embryogenic structures on Schenk and Hildebrand medium without plant growth regulators. Low embryo multiplication rates were obtained when torpedo or early cotyledonary SE were used as initial explant for embryo proliferation, or when glutamine or CH (500 mg L−1) was added to proliferation medium. For germination, cotyledonary-stage SE were isolated and stored at 4 °C for 2 months. After cold storage, SE were cultured on germination medium consisting of Gresshoff and Doy medium, supplemented with 0.44 μM BA and 20 μM silver thiosulphate. Under these conditions, plantlets were regenerated from 21 to 66.7% of the SE generated for both genotypes.

Keywords

Holm oak IAA Leaf explant NAA Shoot apex explant Somatic embryos 
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Notes

Acknowledgements

We thank Dr. Mariano Toribio for kindly providing the plant material used to establish genotypes Q3 and Q10 in vitro. The authors also thank JC Suárez and R Montenegro for their excellent technical assistance.

Funding

This research was partially funded by Ministerio de Economía y Competitividad (MINECO, Spain), through the Projects AGL2013-47400-C4-3-R and AGL2016-76143-C4-4-R.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interests.

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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Group of Biotechnology and Forestry Improvement, Department of Plant PhysiologyInstituto de Investigaciones Agrobiológicas de Galicia, CSICSantiago de CompostelaSpain

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