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Plant Embryo Culture pp 201–213Cite as

Cryopreservation of Zygotic Embryonic Axes and Somatic Embryos of European Chestnut

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 710))

Abstract

For Castanea sativa (European chestnut), a species with recalcitrant seeds that is not easily propagated vegetatively, cryopreservation is one of the most promising techniques for maintaining genetic resource diversity and for conservation of selected germplasms. Long-term conservation of selected seeds and valuable embryogenic lines can be achieved through the cryopreservation of zygotic embryonic axes and somatic embryos, respectively. This chapter describes methods for the desiccation-based cryostorage of zygotic embryonic axes, and the vitrification-based cryopreservation of somatic embryos. For zygotic embryonic axes, the highest post-thaw survival and plantlet recovery rates are obtained by desiccation in a laminar flow hood to 20–25% moisture content, followed by direct immersion in liquid nitrogen. For somatic embryos, embryogenesis resumption rates of over 60% are achieved by preculture of embryo clumps for 3 days on solid medium containing 0.3 M sucrose, incubation in PVS2 vitrification solution for 60 min at 0°C, and direct immersion in liquid nitrogen. Plantlet recovery from cryostored embryogenic lines requires proliferation of the thawed embryos and subsequent maturation before germination and conversion into plantlets.

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References

  1. Conedera M, Manetti MC, Giudici F, Amorini E (2004) Distribution and economic potential of the sweet chestnut (Castanea sativa Mill.) in Europe. Ecol Medit 30:47–61

    Google Scholar 

  2. Bellini E (2005) The chestnut and its resources: images and considerations. Acta Hort 693:85–96

    Google Scholar 

  3. Bounous G (2005) The chestnut: a multipurpose resource for the new millennium. Acta Hort 693:33–40

    Google Scholar 

  4. Pence VC (1995) Cryopreservation of recalcitrant seeds. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 32, Cryopreservation of plant germplasm I. Springer, Berlin, pp 29–50

    Google Scholar 

  5. Vieitez AM, Sánchez MC, García-Nimo ML, Ballester A (2007) Protocol for micropropagation of Castanea sativa. In: Jain SM, Häggman H (eds) Protocols for micropropagation of woody trees and fruits. Springer, Dordrecht, pp 299–312

    Chapter  Google Scholar 

  6. Vieitez FJ, San-José MC, Ballester A, Vieitez AM (1990) Somatic embryogenesis in cultured immature zygotic embryos in chestnut. J Plant Physiol 136:253–256

    Article  Google Scholar 

  7. Vieitez FJ (1995) Somatic embryogenesis in chestnut. In: Jain MS, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants, vol 2. Kluwer Academic, Dordrecht, pp 375–407

    Chapter  Google Scholar 

  8. Sauer U, Wilhelm E (2005) Somatic embryogenesis from ovaries, developing ovules and immature zygotic embryos, and improved embryo development of Castanea sativa. Biol Plant 49:1–6

    Article  Google Scholar 

  9. Corredoira E, Ballester A, Vieitez AM (2006) Somatic embryogenesis in chestnut. In: Mujib S, Samaj J (eds) Somatic embryogenesis, plant cell monograph 2. Springer, Heidelberg, pp 177–199

    Google Scholar 

  10. Corredoira E, Ballester A, Vieitez AM (2003) Proliferation, maturation and germination of Castanea sativa Mill. somatic embryos originated from leaf explants. Ann Bot 92:129–136

    Article  PubMed  CAS  Google Scholar 

  11. Corredoira E, Valladares S, Vieitez AM, Ballester A (2008) Improved germination of somatic embryos and plant recovery of European chestnut. In Vitro Cell Dev Biol Plant 44:307–315

    Article  CAS  Google Scholar 

  12. Corredoira E, Montenegro D, San-José MC, Vieitez AM, Ballester A (2004) Agrobacterium-mediated transformation of European chestnut embryogenic cultures. Plant Cell Rep 23:311–318

    Article  PubMed  CAS  Google Scholar 

  13. Corredoira E, San-José MC, Vieitez AM, Ballester A (2007) Improving genetic transformation of European chestnut and cryopreservation of transgenic lines. Plant Cell Tissue Organ Cult 91:281–288

    Article  CAS  Google Scholar 

  14. Maynard CA, Powell WA, Polin-McGuigan LD, Vieitez AM, Ballester A, Corredoira E, Merkle SA, Andrade GM (2008) Chestnut. In: Kole C, Hall TC (eds) Compendium of transgenic crop plants: transgenic forest tree species. Blackwell, Oxford, pp 169–192

    Google Scholar 

  15. Engelmann F (2000) Importance of cryopreservation for the conservation of plant genetic resources. In: Engelmann F, Takagi H (eds) Cryopreservation of tropical plant ­germplasm – current research progress and application. JIRCAS, Tsukuba/IPGRI, Rome, pp 8–20

    Google Scholar 

  16. Lambardi M, De Carlo A (2003) Application of tissue culture to the germplasm conservation of temperate broad-leaf trees. In: Jain SM, Ishii K (eds) Micropropagation of woody trees and fruits. Kluwer Academic, Dordrecht, pp 815–840

    Chapter  Google Scholar 

  17. Klimaszewska K, Cyr DR (2002) Conifer somatic embryogenesis: I. Development. Dendrobiology 48:31–39

    Google Scholar 

  18. Vendrame WA, Holliday CP, Montello PM, Smith DR, Merkle SA (2001) Cryopreservation of yellow-poplar and sweetgum embryogenic cultures. New Forests 21:283–292

    Article  Google Scholar 

  19. Pence VC (1992) Desiccation and the survival of Aesculus, Castanea and Quercus embryo axes through cryopreservation. Cryobiology 29:391–399

    Article  Google Scholar 

  20. Corredoira E, San-José MC, Ballester A, Vieitez AM (2004) Cryopreservation of zygotic embryo axes and somatic embryos of European chestnut. CryoLetters 25:33–42

    PubMed  Google Scholar 

  21. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  22. Sakai A, Kobayashi S, Oiyama I (1990) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. Var. brasiliensis Tanaka) by vitrification. Plant Cell Rep 9:30–33

    Article  Google Scholar 

  23. Poulsen KM (1992) Sensitivity to desiccation and low temperatures (−196°C) of embryo axes from acorns of the pedunculate oak (Quercus robur L.). CryoLetters 13:75–82

    Google Scholar 

  24. Sakai A (1995) Cryopreservation of ­germplasm of woody plants. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 32, Cryopreservation of plant germplasm I. Springer, Berlin, pp 53–69

    Google Scholar 

  25. San-José MC, Ballester A, Vieitez AM (2001) Effect of thidiazuron on multiple shoot induction and plant regeneration from cotyledonary nodes of chestnut. J Hortic Sci Biotech 76:588–595

    Google Scholar 

  26. Valladares S, Toribio M, Celestino C, Vieitez AM (2004) Cryopreservation of embryogenic cultures from mature Quercus suber trees using vitrification. CryoLetters 25:177–186

    PubMed  Google Scholar 

  27. Yeung EC (1995) Structural and developmental patterns in somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Kluwer Academic, Dordrecht, pp 205–247

    Chapter  Google Scholar 

  28. Holliday C, Merkle S (2000) Preservation of American chestnut germplasm by cryostorage of embryogenic cultures. J Am Chestnut Found 14:46–52

    Google Scholar 

  29. Andrade GM, Merkle SA (2005) Enhancement of American chestnut somatic seedling production. Plant Cell Rep 24:326–334

    Article  PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Instituto de Investigaciones Agrobiológicas de Galicia, CSIC, Santiago de Compostela, Spain

    Ana M. Vieitez, M. Carmen San-José & Elena Corredoira

Authors
  1. Ana M. Vieitez
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  2. M. Carmen San-José
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  3. Elena Corredoira
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Corresponding author

Correspondence to Ana M. Vieitez .

Editor information

Editors and Affiliations

  1. Dept. Biological Sciences, University of Calgary, University Dr. NW., 2500, Calgary, T2N 1N4, Alberta, Canada

    Trevor A. Thorpe

  2. , Department of Biological Sciences, University of Calgary, University Dr. NW 2500, Calgary, T2N 1N4, Alberta, Canada

    Edward C. Yeung

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© 2011 Humana Press

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Vieitez, A.M., San-José, M.C., Corredoira, E. (2011). Cryopreservation of Zygotic Embryonic Axes and Somatic Embryos of European Chestnut. In: Thorpe, T., Yeung, E. (eds) Plant Embryo Culture. Methods in Molecular Biology, vol 710. Humana Press. https://doi.org/10.1007/978-1-61737-988-8_15

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  • DOI: https://doi.org/10.1007/978-1-61737-988-8_15

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61737-987-1

  • Online ISBN: 978-1-61737-988-8

  • eBook Packages: Springer Protocols

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