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Cryopreservation of Hybrid Pinus elliottii × P. caribaea

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Step Wise Protocols for Somatic Embryogenesis of Important Woody Plants

Part of the book series: Forestry Sciences ((FOSC,volume 84))

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Abstract

The cryopreservation of embryogenic cultures within operational forestry proves to be a crucial tool, to store clones without loss of juvenility, while field tests, to identify genotypes with significant genetic gain, are being conducted. Besides several conifers species have been cryopreserved till now, only few studies have been reported for hybrid species. The interspecific pine hybrid, P. elliottii var. elliottii x P. caribaea var. hondurensis has a major economic importance mainly in South America, West Africa and Australia. The hybrid superiority appears to be derived from a complementary recombination of traits from the two parental species–growth rate and high yield of resin production from P. caribaea var. hondurensis, combined with wind-firmness, adaptability to wet sites, high wood-density and stem straightness of P. elliottii var. elliottii. This chapter describes a complete cryopreservation procedure for embryonal mass of the hybrid P.elliottii var. elliottii x P. caribaea var. hondurensis, and a methodology to analyse the ploidy stability by flow cytometry.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

ABA :

Abscisic acid

BAP :

6-benzylaminopurine

DMSO :

Dimethyl sulfoxide

ECL:

Embryogenic cell line

EM:

Embryonal mass

mLV:

Modified Litvay’s medium

MVF:

Multi-varietal forestry

mLV:

Modified Litvay medium

PEG:

Polyethylene glycol

Pi:

Propidium iodide

PGR:

Plant growth regulators

SE:

Somatic embryogenesis

PEE:

Pinus elliottii var. elliottii

PCH:

Pinus caribaea var. hondurensis

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Acknowledgements

This research was suported by CENTRO-07-0202-FEDER-018579- “I & D em tecnologias e técnicas de clonagem ‘in vitro’, micropropagação e clonagem de plantas e genótipos”, co-funded by QREN, “Programa Mais Centro-Operacional Regional do Centro”, EU through European Regional Development Fund (FEDER), and FCT/MEC and FEDER, PT2020 Partnership Agreement & COMPETE 2020; POCI/01/0145/FEDER/007265, UID/QUI/50006/2013, UID/BIA/04004/2013, UID/QUI/00062/2013. F.C.T. funded M.C. Dias fellowship SFRH/BPD/100865/2014. Authors thank to Eng. Cláudio Pinheiro from Brazilian company Resisul for the supply of seeds and to Klon’s technicians D. Sousa, J. Figueiredo and N. Mano for technical support.

Author information

Authors and Affiliations

  1. KLÓN—Innovative Technologies from Cloning, S.A., Biocant Park, 3060-197, Cantanhede, Portugal

    Liliana Marum, Sandra Nunes, Tânia Almeida, Vanessa Tolentino Pereira & Nelson Farinha

  2. Department of Life Science, Centre for Functional Ecology (CFE), University of Coimbra, Calçada Martim de Freitas, 3000-456, Coimbra, Portugal

    Maria Celeste Dias

  3. Department of Chemistry and QOPNA, University of Aveiro, 3810-193, Aveiro, Portugal

    Maria Celeste Dias

  4. Department of Biology & LAQV/REQUIMTE, Faculty of Sciences, University of Porto, Rua do Campo Alegre, 4169-007, Porto, Portugal

    Conceição Santos

  5. Centro de Biotecnologia Agrícola e Agro-Alimentar do Alentejo (CEBAL), Instituto Politécnico de Beja (IPBeja), 7801-908, Beja, Portugal

    Liliana Marum

  6. Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Universidade de Évora, Pólo da Mitra, 7002-554, Évora, Portugal

    Liliana Marum

Authors
  1. Liliana Marum
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  2. Sandra Nunes
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  3. Tânia Almeida
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  4. Vanessa Tolentino Pereira
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  5. Nelson Farinha
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  6. Maria Celeste Dias
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  7. Conceição Santos
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Corresponding author

Correspondence to Liliana Marum .

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Editors and Affiliations

  1. Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland

    Shri Mohan Jain

  2. Technology Center, Weyerhaeuser Company, Federal Way, Washington, USA

    Pramod Gupta

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Marum, L. et al. (2018). Cryopreservation of Hybrid Pinus elliottii × P. caribaea. In: Jain, S., Gupta, P. (eds) Step Wise Protocols for Somatic Embryogenesis of Important Woody Plants. Forestry Sciences, vol 84. Springer, Cham. https://doi.org/10.1007/978-3-319-89483-6_21

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