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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
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.
Hosie, R.C. (ed.) (1979) Native Trees of Canada. 8th ed. Fitzhenry and Whiteside, Don Mills.
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.
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.
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.
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.
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.
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.
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.
von Arnold, S. and Eriksson, T. (1981) In vitro studies on adventitious shoot formation in Pinus contorta. Can. J. Bot. 59, 870–874.
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.
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.
Tremblay, F.M. (1990) Somatic embryogenesis and plantlet regeneration from embryos isolated from stored seeds of Picea glauca. Can. J. Bot. 68, 236–242.
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.
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.
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.
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.
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.
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.
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.
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.
Kong, L. (1994) Factors affecting white spruce somatic embryogenesis and embryo conversion. Ph.D. Dissertation, University of Calgary, Calgary.
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
Editor information
Editors and Affiliations
Rights 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