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Cryopreservation of Hybrid Pinus elliottii × P. caribaea

  • Liliana Marum
  • Sandra Nunes
  • Tânia Almeida
  • Vanessa Tolentino Pereira
  • Nelson Farinha
  • Maria Celeste Dias
  • Conceição Santos
Chapter
Part of the Forestry Sciences book series (FOSC, volume 84)

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.

Keywords

Cryopreservation Cryoprotectors Pine hybrid P. elliottii × P. caribaea Genome stability Ploidy level 

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

Notes

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.

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Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  • Liliana Marum
    • 1
    • 5
    • 6
  • Sandra Nunes
    • 1
  • Tânia Almeida
    • 1
  • Vanessa Tolentino Pereira
    • 1
  • Nelson Farinha
    • 1
  • Maria Celeste Dias
    • 2
    • 3
  • Conceição Santos
    • 4
  1. 1.KLÓN—Innovative Technologies from CloningCantanhedePortugal
  2. 2.Department of Life ScienceCentre for Functional Ecology (CFE), University of CoimbraCoimbraPortugal
  3. 3.Department of Chemistry and QOPNAUniversity of AveiroAveiroPortugal
  4. 4.Department of Biology & LAQV/REQUIMTE, Faculty of SciencesUniversity of PortoPortoPortugal
  5. 5.Centro de Biotecnologia Agrícola e Agro-Alimentar do Alentejo (CEBAL), Instituto Politécnico de Beja (IPBeja)BejaPortugal
  6. 6.Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Universidade de ÉvoraÉvoraPortugal

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