Skip to main content

Somatic Embryogenesis in Picea Suspension Cultures

  • Protocol
  • 2712 Accesses

Part of the Methods in Molecular Biology™ book series (MIMB,volume 318)

Abstract

Generation of somatic embryos in spruce is achieved through the execution of five steps designated as: (1) induction of embryogenic tissue, (2) maintenance of embryogenic tissue, (3) embryo development, (4) embryo maturation, and (5) conversion into plants. Depending on species and genotypes within the same species, each step must be optimized for obtaining maximum results. In general, embryogenic tissue is generated from immature and mature zygotic embryos and maintained in either liquid or solid conditions in the presence of plant growth regulators auxin and cytokinin. Initiation of embryo development in suspension cultured is induced by removal of plant growth regulators, whereas continuation of development and completion of maturation require applications of abscisic acid and imposition of a desiccation period. Both treatments are needed for conferring morphological and physiological maturation to the embryos. Mature somatic embryos are germinated in the absence of plant regulators and embryo conversion (i.e., formation of a functional shoot and root, occurs after a few weeks in culture).

Key Words

  • Conversion
  • drying treatment
  • embryogenic tissue
  • embryo development and maturation
  • germination
  • spruce

This is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1385/1-59259-959-1:087
  • Chapter length: 13 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   139.00
Price excludes VAT (USA)
  • ISBN: 978-1-59259-959-2
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   179.00
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)
Fig. 1.

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Thorpe, T.A. and Stasolla, C. (2001) Somatic embryogenesis, in Current Trends in the Embryology of Angiosperms (Bhojwani, S.S., ed.), Kluwer Acad. Publisher, Dordrecht. pp. 35–64.

    Google Scholar 

  2. Hosie, R.C. (ed.) (1979) Native Trees of Canada. 8th ed. Fitzhenry and Whiteside, Don Mills.

    Google Scholar 

  3. Durzan, D.J. (1980) Progress and promise in forest genetics. In: proceedings of the 50th Anniversary Conference, Paper Science and Technology, The Cutting Edge, May 8–10, pp 31–60. The Institute of Paper Chemistry, Appleton, WI.

    Google Scholar 

  4. Lu, C. Y. and Thorpe, T. A. (1987) Somatic embryogenesis and plantlet regeneration in cultured immature embryos of Picea glauca. J. Plant Physiol. 128, 297–302.

    CAS  Google Scholar 

  5. Hakman, I., Rennie, P., and Fowke, L.C. (1987) A light and electron microscopy study of Picea glauca (white spruce) somatic embryos. Protoplasma 140, 100–109.

    CrossRef  Google Scholar 

  6. Von Arnold, S., Sabala, I., Bozhkov, P., Dyachok, J., and Filonova, L. (2002) Developmental pathways of somatic embryogenesis. Plant Cell Tiss. Org. Cult. 69, 233–249.

    CrossRef  Google Scholar 

  7. Harry, I.S. and Thorpe, T.A. (1991), Somatic embryogenesis and plant regeneration from mature zygotic embryos of red spruce. Bot. Gaz. 152, 446–445.

    CAS  CrossRef  Google Scholar 

  8. El Meskaoui, A. and Tremblay, F. (2001) Involvement of ethylene in the maturation of black spruce embryogenic cell lines with different maturation capacities. J. Exp. Bot. 52, 761–769.

    PubMed  Google Scholar 

  9. Bozhkov, P.V. and von Arnold, S. (1998) Polyethylene glycol promotes maturation but inhibits further development of Picea abies somatic embryos. Physiol. Plant. 104, 211–224.

    CAS  CrossRef  Google Scholar 

  10. von Arnold, S. and Eriksson, T. (1981) In vitro studies on adventitious shoot formation in Pinus contorta. Can. J. Bot. 59, 870–874.

    CrossRef  Google Scholar 

  11. Joy IV, R.W., Yeung, E.C., Kong, L. and Thorpe, T.A. (1991) Development of white spruce somatic embryos: I. Storage product deposition. In Vitro Cell. Dev. Biol. Plant 27P, 32–41.

    CAS  Google Scholar 

  12. Litvay, J.D., Johnson, M.A., Verma, D., Einspahr, D., and Weyrauch, K. (1985) Conifer suspension culture medium development using analytical data from developing seeds. Institute Paper Chem. Tech. Paper Series 155, Appleton, Wisconsin.

    Google Scholar 

  13. Tremblay, F.M. (1990) Somatic embryogenesis and plantlet regeneration from embryos isolated from stored seeds of Picea glauca. Can. J. Bot. 68, 236–242.

    CAS  CrossRef  Google Scholar 

  14. Attree, S.M., Budimir, S., and Fowke, L.C. (1990) Somatic embryogenesis and plantlet regeneration from cultured shoots and cotyledons of seedlings from stored seeds of black and white spruce (Picea mariana and Picea glauca). Can. J. Bot. 68, 30–34.

    CAS  CrossRef  Google Scholar 

  15. Filonova, L. H, Bozhkov, P. V., and von Arnold, S. (2000) Developmental pathway of somatic embryogenesis in Picea abies as revealed by time-lapse tracking. J. Exp. Bot. 51, 249–264.

    CAS  CrossRef  PubMed  Google Scholar 

  16. Bozhkov, P.V., Filonova, L.H., and von Arnold, S. (2002) A key developmental switch during Norway spruce somatic embryogenesis is induced by withdrawal of plant growth regulators and is associated with cell death and extracellular acidification. Biotech. Bioen. 77, 658–667.

    CAS  CrossRef  Google Scholar 

  17. Kong, L. and Yeung, E.C. (1992) Development of white spruce somatic embryos: II. Continual shoot meristem development during germination. In Vitro Cell. Dev. Biol. Plant 28P, 125–131.

    Google Scholar 

  18. Stasolla, C. and Yeung, E.C. (1999) Ascorbic acid improves the conversion of white spruce somatic embryos. In Vitro Cell. Dev. Biol. Plant 35, 316–319.

    CAS  CrossRef  Google Scholar 

  19. Stasolla, C., Kong, L., Yeung, E.C., and Thorpe, T.A. (2002). Maturation of somatic embryos in conifers: morphogenesis, physiology, biochemistry, and molecular biology. In Vitro Cell. Dev. Biol-Plant 38, 93–105.

    CAS  CrossRef  Google Scholar 

  20. Attree, S.M. and Fowke, L.C. (1993) Embryogeny of gymnosperms: advances in synthetic seed technology of conifers. Plant Cell Tiss. Org. Cult. 35, 1–35.

    CAS  CrossRef  Google Scholar 

  21. Roberts, D.R., Sutton, B.C.S., and Flinn, B.S. (1990) Synchronous and high frequency germination of interior spruce somatic embryos following partial drying at high relative humidity. Can. J. Bot. 68, 1086–1090.

    CrossRef  Google Scholar 

  22. Kong, L. (1994) Factors affecting white spruce somatic embryogenesis and embryo conversion. Ph.D. Dissertation, University of Calgary, Calgary.

    Google Scholar 

Download references

Acknowledgments

The author acknowledges with gratitude the support received from the Natural Sciences and Engineering Research Council of Canada in the form of a Discovery Grant.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

© 2006 Humana Press Inc.

About this protocol

Cite this protocol

Stasolla, C. (2006). Somatic Embryogenesis in Picea Suspension Cultures. In: Loyola-Vargas, V.M., Vázquez-Flota, F. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology™, vol 318. Humana Press. https://doi.org/10.1385/1-59259-959-1:087

Download citation

  • DOI: https://doi.org/10.1385/1-59259-959-1:087

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-547-7

  • Online ISBN: 978-1-59259-959-2

  • eBook Packages: Springer Protocols