Necking in Embryonal Tube Cells and Its Implications for Morphogenic Protoplasts and Conifer Tree Improvement

  • Don J. Durzan
Part of the NATO ASI Series book series (NSSA, volume 210)


The breakup of viscous protoplasm into alternate smaller and larger protoplasts (necking) occurs naturally as a product of stress in elongated cells of an embryonal-suspensor mass. Necking can also be induced as an artifact of handling these actively streaming cells. Protoplasts, formed as a product of necking inside cells, can be released and recovered by cell-wall digesting enzymes. At least eight size classes of protoplasts can be recovered from cell suspension cultures of the embryonal-suspensor mass. This range of size classes is based on the different cell types (proembryonal, embryonal tube, embryonal suspensor, upper suspensor) and by limited spontaneous fragmentation and fusion of protoplasts among recovered size classes. Staining properties among recovered protoplast size classes reveal that: i) those derived from proembryonal cells may have morphogenic potential; ii) more than one protoplast inside cells of the embryonal tube and suspensor may contain a nucleus derived from a free nuclear stage; iii) the protoplast population represents a varied and fractional genetic potential based on organelles trapped in protoplasts during the necking and fusion processes; and iv) necking in conifer cells may explain illustrations in the literature showing multiple migrating nuclei just after fertilization or in cylindrical cells of the embryonal-suspensor mass. These nuclei produce a new cytoplasm or neocytoplasm associated with the establishment of the classical “basal plan” for proembryonal development. This neocytoplasm interacts in unknown ways to select for chloroplast genomes from pollen and mitochondria of the female parent.


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  1. 1.
    Adamson, A.W., 1982, “Physical Chemistry of Surfaces,” 4th Edition. J. Wiley and Sons, N.Y.Google Scholar
  2. 2.
    Allen, G.S. and Owens, J.N., 1972, “The Life History of Douglas Fir.” Environment Canada, Forestry Service, Ottawa.Google Scholar
  3. 3.
    Attree, S.M., Dunstan, D.I. and Fowke, L.C., 1989, Plantlet regeneration from embryogenic protoplasts of white spruce (Picea glauca). Bio/Tech. 7:1060–1062.Google Scholar
  4. 4.
    Batchelor, G.K., 1967, “An Introduction to Fluid Dynamics.” Cambridge University Press, Cambridge.Google Scholar
  5. 5.
    Bold, H.C., Alexopoulos, C.J., and Delevoryas, T., 1980, “Morphology of Plants and Fungi.” Harper and Row, New York.Google Scholar
  6. 6.
    Boucher, E.A., 1980, Capillary phenomena: properties of systems with fluid/fluid interfaces, Rep. Prog. Phys. 43:497–546.CrossRefGoogle Scholar
  7. 7.
    Boulay, M.P., Gupta, P.K., Krogstrup, P. and Durzan, D.J., 1988, Development of somatic embryos from cell suspension culture of Norway spruce (Picea abies Karst), Plant Cell Reports 7:134–137.CrossRefGoogle Scholar
  8. 8.
    Bradley, W.H., 1965, Vertical density currents. Science 150:1423–1428.CrossRefGoogle Scholar
  9. 9.
    Camefort, H., 1969, Fécondation et proembryogénèse chez les Abietaceés (notion de neocytoplasme), Rev. Cytol. Biol. Vég. 32:253–2711.Google Scholar
  10. 10.
    Chamberlain, C.J., 1935, “Gymnosperms. Structure and Evolution.” University of Chicago Press, Chicago.Google Scholar
  11. 11.
    Dawkins, R., 1981, “The Extended Phenotype,” W.H. Freeman and Co., San Francisco.Google Scholar
  12. 12.
    Dogra, P.D., 1966, Observations on Abies pindrow with a discussion on the question of occurence of apomixis in gymnosperms. Silvae Genetica 15:1–32.Google Scholar
  13. 13.
    Dogra, P.D., 1980, Embryogeny of gymnosperms and taxonomy — an assessment. In: “Glimpses in Plant Research,” P.K.K. Nair, ed., Vikas House Ltd., New Delhi, 5:114–128.Google Scholar
  14. 14.
    Dogra, P.D., 1978, Morphology, development and nomenclature of conifer embryo, Phytomorphology 28:307–322.Google Scholar
  15. 15.
    Dogra, P.D., 1967, Seed sterility and disturbances in embryogeny in conifers with particular reference to seed testing and tree breeding in Pinaceae. Stud. Forestal. Suecica 45:1–97.Google Scholar
  16. 16.
    Doyle, J., 1963, Proembryogeny in Pinus in relation to that in other conifers — a survey. Proc. Roy. Irish Acad. 62B: 181–216.Google Scholar
  17. 17.
    Durzan, D.J., 1988, Process control in somatic polyembryogenesis. In: “Molecular Genetics of Forest Trees,” Franz Kempe Symp., June 14–16, 1988, Swedish Univ. Agric. Sci, Umea, Rept. No. 8, 147–186.Google Scholar
  18. 18.
    Durzan, D.J., 1988, Somatic polyembryogenesis for the multiplication of tree crops, Biotech Genetic Eng. Revs. 6:339–376.Google Scholar
  19. 19.
    Durzan, D.J. and Gupta, P.K., 1987, Somatic embryogenesis and polyembryogenesis in Douglas fir cell suspension cultures, Plant Sci. 52:229–235.CrossRefGoogle Scholar
  20. 20.
    Eberhard, W.G., 1980, Evolutionary consequences of intracellular organelle competition, Quart. Rev. Biol. 55:231–249.CrossRefPubMedGoogle Scholar
  21. 21.
    Gupta, P.K., Dandekar, A.M. and Durzan, D.J., 1988, Somatic proembryo formation and transient expression of luciferase gene in Douglas fir and loblolly pine protoplasts. Plant Sci. 58:85–92.CrossRefGoogle Scholar
  22. 22.
    Gupta, P.K. and Durzan, D.J., 1987a, Somatic embryos from protoplasts of loblolly pine proembryonal cells. Bio/Tech. 5:710–712.Google Scholar
  23. 23.
    Gupta, P.K. and Durzan, D.J., 1987b, Biotechnology of somatic polyembryogenesis and plantlet regeneration in loblolly pine. Bio/Tech. 5:147–151.Google Scholar
  24. 24.
    Gupta, P.K. and Durzan, D.J., 1987c, Plantlet regeneration via somatic embryogenesis from subculture callus of mature embryos of Picea abies (Norway spruce). In Vitro Cell, and Devel. Biol. 22:685–688.CrossRefGoogle Scholar
  25. 25.
    Haissig, B.E., Nelson, N.D. and Kidd, G.H., 1987, Trends in the use of tissue culture in forest improvement. Bio/Tech. 5:52–57.Google Scholar
  26. 26.
    Holliday, R., 1987, The inheritance of epigenetic defects. Science 238:163–170.CrossRefPubMedGoogle Scholar
  27. 27.
    Israelachvil, J.N. and McGuiggan, P.M., 1988, Forces between surfaces in liquids. Science 241:795–800.CrossRefGoogle Scholar
  28. 28.
    Kay, J.M. and Nedderman, R.M., 1985, “Fluid Mechanics and Transfer Processes.” Cambridge Univ. Press, Cambridge. 28. Klekowski, E.J. Jr., 1988, “Mutation, Developmental Selection, and Plant Evolution.” Columbia Univ. Press, New York.Google Scholar
  29. 30.
    Lörz, H., 1984, Enucleation of protoplasts: Preparation of cytoplasts and miniprotoplasts. In: “Cell Culture and Somatic Cell Genetics of Plants,” Vol. 1, “Laboratory Procedures and Their Applications,” I.K. Vasil, ed., Academic Press, New York, p. 448–453.Google Scholar
  30. 31.
    Neale, D.B. and Sederoff, R., 1988, Inheritance and evolution of conifer organelle genomes. In: “Genetic Manipulation of Woody Plants,” J.W. Hanover and D.E. Keathley, eds., Plenum Press, pp. 251–264.CrossRefGoogle Scholar
  31. 32.
    Powledge, T.M., 1984, Biotechnology touches the forest. Bio/Tech. 2:763–772.Google Scholar
  32. 33.
    Singh, H., 1978, “Embryology of Gymnosperms.” Enc. Plant Physiol., Vol. 10, Part 2. Gebrüder, Borntraeger, Berlin.Google Scholar
  33. 34.
    Sinnott, E.W., 1960, “Plant Morphogenesis.” McGraw-Hill Inc., New York.CrossRefGoogle Scholar
  34. 35.
    Sziklai, O., 1986, Polyembryony of Pinus contorta Doug. in central Yukon. In: “Provenances and Forest Tree Breeding for High Latitudes,” D. Lindgren, ed., Proc. F. Kempe Symp., Swedish Univ. Agric. Sci., Umea, Rept. 6., p. 251.Google Scholar
  35. 36.
    Tanaka, T., Nishio, I., Sun, S.-T. and Ueno-Nishio, S., 1982, Collapse of gels in an electric field. Science 218:467–469.CrossRefPubMedGoogle Scholar
  36. 37.
    Tanksley, S.D., Young, N.D., Paterson, A.H. and Bonierbale, M.W., 1989, RFLP mapping in plant breeding. New tools for an old science. Bio/Tech. 7:257–264.Google Scholar
  37. 38.
    Thompson, D.W., 1963, “On Growth and Form,” Vol. 1. Cambridge Univ. Press, Cambridge.Google Scholar
  38. 39.
    Vicsek, T., 1989, “Fractal Growth Phenomena,” World Scientific, Singapore.CrossRefGoogle Scholar
  39. 40.
    Vollrath, F. and Edmonds, D.T., 1989, Modulation of the mechanical properties of spider silk by coating with water. Nature 340:305–307.CrossRefGoogle Scholar
  40. 41.
    Widolm, J.M., 1972, The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Tech. 47:189–194.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Don J. Durzan
    • 1
  1. 1.Department of Environmental HorticultureUniversity of CaliforniaDavisUSA

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