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Somatic embryogenesis in Pinus halepensis Mill.: an important ecological species from the Mediterranean forest

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Abstract

Pinus halepensis Mill. is a common forest species in the Mediterranean area and it is important for environmental conservation. This study established a method of regenerating Pinus halepensis Mill. through somatic embryogenesis. The effect of culture medium (mineral salts, nitrogen source and plant growth regulators), collection date and seed family on embryogenic tissue initiation and proliferation in Pinus halepensis was analysed during the first steps of embryogenesis process. This study showed a marked effect of the culture medium tested as well as some significant differences among collection dates. Furthermore, the embryogenic tissue initiation was affected by the amino acid mixture in the culture medium and the proliferation stage was significantly affected by the combination of plant growth regulators. At the end of the maturation phase the presence of activated charcoal was also evaluated. Finally, maturation of embryogenic tissue was affected by the nitrogen source in the culture medium and these results were different for high and low mature embryo producing cell lines. To the best of our knowledge, this is the first report on Aleppo pine somatic embryogenesis describing a simple and efficient procedure for large-scale somatic embryo production.

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Abbreviations

A:

Maturation medium, DCR basal medium supplemented with: 75 μM ABA, 9 g L−1 of Gelrite®, 4.5 % (w/v) sucrose, and ED amino acid mixture

ABA:

Abscisic acid

AC:

Activated charcoal

B:

Maturation medium, DCR medium supplemented with: 75 μM ABA and 9 g L−1 of Gelrite®, 4.5 % (w/v) sucrose and ED amino acid mixture with 1,650 mg L−1 of glutamine

BA:

N6-benzyladenine

C:

Maturation medium, DCR basal medium supplemented with: 75 μM ABA, 9 g L−1 of Gelrite®, 6 % (w/v) sucrose and ED amino acid mixture

D:

Maturation medium, DCR medium supplemented with: 75 μM ABA and 9 g L−1 of Gelrite®, 6 % (w/v) sucrose and ED amino acid mixture with 1,650 mg L−1 glutamine

2,4-D:

2,4-Dichlorophenoxyacetic acid

DBA:

9 μM 2,4D and 2.7 μM BA

DCR:

Gupta and Durzan basal medium (Gupta and Durzan 1985)

DKI:

9 μM 2,4D and 2.7 μM Kinetin

DNB:

4.5 μM 1-Naphthaleneacetic acid, 4.5 μM 2,4D and 2.7 μM BA

ECLs:

Established cell lines

ED:

EDM amino acid mixture

EDM:

Embryo development medium (Walter et al. 1998)

ET:

Embryogenic tissue

FW:

Fresh weight

LP:

Quorin and Lepoivre medium (Quoirin and Lepoivre 1977, modified by Aitken-Christie et al. 1988)

SE:

Somatic embryogenesis

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Acknowledgments

We thank Charlie B. Low for his statistical advice. This research was funded by Ministerio de Ciencia y Tecnología—Spain (AGL2005-08214-CO2-02) and Departamento de Agricultura y Pesca-Gobierno Vasco (VEC2004021 and VED2007014), who granted I. A. M. with a PhD scholarship.

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Authors and Affiliations

  1. Centro de Arkaute, Neiker-Tecnalia, Apdo. 46, 01080, Vitoria-Gasteiz, Spain

    I. A. Montalbán, A. Setién-Olarra & P. Moncaleán

  2. Scion, Private Bag 3020, Rotorua, 3046, New Zealand

    C. L. Hargreaves

Authors
  1. I. A. Montalbán
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  2. A. Setién-Olarra
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  3. C. L. Hargreaves
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  4. P. Moncaleán
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Corresponding author

Correspondence to P. Moncaleán.

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Communicated by K. Klimaszewska.

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Montalbán, I.A., Setién-Olarra, A., Hargreaves, C.L. et al. Somatic embryogenesis in Pinus halepensis Mill.: an important ecological species from the Mediterranean forest. Trees 27, 1339–1351 (2013). https://doi.org/10.1007/s00468-013-0882-0

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