Plant Cell, Tissue and Organ Culture

, Volume 95, Issue 1, pp 69–78 | Cite as

Factors affecting maintenance, proliferation, and germination of secondary somatic embryos of Eucalyptus globulus Labill

  • Gloria Pinto
  • Yill-Sung Park
  • Sónia Silva
  • Lucinda Neves
  • Clara Araújo
  • Conceição Santos
Original Paper


The described protocol for repetitive somatic embryogenesis (SE) in Eucalyptus globulus produced more somatic embryos than the primary SE protocol. Primary somatic embryos (induced on MS3NAA) were transferred to the same medium, leading to new cycles of somatic embryos, for at least 2 years. The influence of medium (MS and B5), plant growth regulators (auxins and cytokinins), and light on secondary SE was tested. The MS medium without growth regulators (MSWH) was more efficient for cotyledonary embryo formation and germination than the B5 medium. Reducing auxin (NAA) levels increased the proliferation of globular somatic embryos and allowed SE competence to be maintained on medium free of plant growth regulators. The addition of two cytokinins (BAP and KIN) to the MS medium did not improve proliferation of globular secondary embryos, but was crucial during later stages of the SE process (germination and conversion). Data also show that light may influence the quality of the process, depending on its stage. Darkness should be maintained until the cotyledonary stage is reached, after which exposure to light is recommended.


Embryogenic competence Eucalyptus Growth regulators Media composition Secondary somatic embryogenesis 



Gamborg medium






Murashige and Skoog medium


α-Naphthalene acetic acid


MS medium without growth regulators


Somatic embryogenesis


Zygotic embryos


Plant growth regulators



Authors thank Celbi for providing the material used in this study and for supporting part of the project. The Portuguese Foundation for Science and Technology FCT/MCT project POCI/AGR/60672/2004 also supported part of the project. Thanks are also due to José Dias and Armando Costa for technical assistance. FCT supported Glória′s (FCT/SFRH/BPD/26434/2006), Sónia′s (SFRH/BD/32257/2006) fellowships.


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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Gloria Pinto
    • 1
  • Yill-Sung Park
    • 2
  • Sónia Silva
    • 1
  • Lucinda Neves
    • 3
  • Clara Araújo
    • 3
  • Conceição Santos
    • 1
  1. 1.Department of Biology, Laboratory of Biotechnology and Cytomics, CESAMUniversity of AveiroAveiroPortugal
  2. 2.Natural Resources Canada, Canadian Forestry Service – Canadian Wood Fibre CentreFrederictonCanada
  3. 3.Silvicaima SA Constância SulConstanciaPortugal

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