Summary
The growth and development of white spruce somatic embryos was followed from the filamentous immature to the mature cotyledonary embryo stage. Histochemical examination of the various stages of embryo development showed that lipids, proteins, and polysaccharides were produced to varying degrees during the process. During early stages (1 to 2 wk on ABA), mostly polysaccharide was produced, whereas during later stages, polysaccharides, lipids, and protein accumulated. Electron microscopy indicated that lipid deposition in somatic embryos started during the first week after transfer to ABA-containing medium. Deposition of the storage products began at the basal end of the embryonal mass and within the proximal zone of the suspensors. Accumulation continued to the peripheral regions and then inward toward the cortex of the developing embryo. In all cases, polysaccharide accumulated first, followed by lipid and lastly, protein. Quantitatively, cotyledonary stage somatic embryos had less lipid and protein and more starch when compared to zygotic embryos at the same developmental stage. Total protein profiles elucidated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the majority of proteins were similar in zygotic and somatic embryos. Prominent protein bands were found at 30, 20, 19.5, 15, 14.4, 12, and 10 Kd. However, protein bands at 40, 15, and 12 Kd in total protein from somatic embryos were either absent or highly underexpressed.
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References
Buchholz, J. T. Suspensor and early embryo of pines. Bot. Gaz. 66:185–233; 1918.
Clutter, M.; Brady, T.; Walbot, V., et al. Macromolecular synthesis during plant embryogeny. Cellular rates of RNA synthesis in diploid and polytene cells in bean embryos. J. Cell Biol. 63:1097–1102; 1974.
Dubois, M.; Billes, K. A.; Hamilton, J. K., et al. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28:350–356; 1956.
Dunstan, D. I. Prospects and progress in conifer biotechnology. Can. J. For. Res. 18:1497–1506; 1988.
Durzan, D. J. Physiological states and metabolic phenotypes in embryonic development. In: Bonga, J. M.; Durzan, D. J., eds. Cell and tissue culture in Forestry, vol. 2, Boston, Dordrecht; Martinus-Nijhoff Pubs. 1987:405–439.
Feirer, R. P.; Conkey, J. H.; Verhagen, S. A. Triglycerides in embryogenic conifer calli: a comparison with zygotic embryos. Plant Cell Rep. 8:207–209; 1989.
Finkelstein, R. R.; Crouch, M. L. Rapeseed embryo development in culture on high osmoticum is similar to that in seeds. Plant Physiol. 81:907–912; 1986.
Finkelstein, R. R.; Somerville, C. Abscisic acid or high osmoticum promote accumulation of long-chain fatty acids in developing embryos ofBrassica napus. Plant Science 61:213–217; 1989.
Gifford, D. J.; Tolley, M. C. The seed proteins of white spruce and their mobilization following germination. Physiol. Plant. 77:254–261; 1989.
Hakman, I. Somatic embryogenesis in Norway spruce (Picea abies). 4th International Conifer Tissue Culture Work Group, Aug. 8–12, 1988, Saskatoon, Sask., Canada, Abstr.
Hakman, I.; Rennie, P.; Fowke, L. A light and electron microscope study ofPicea glauca (white spruce) somatic embryos. Protoplasma 140:100–109; 1987.
Hakman, I.; Fowke, L. C. Somatic embryogenesis inPicea glauca (white spruce) andPicea mariana (black spruce). Can. J. Bot. 65:656–659; 1987.
Hakman, I.; von Arnold, S. Somatic embryogenesis and plant regeneration from suspension cultures onPicea glauca (white spruce). Physiol. Plant. 72:579–587; 1988.
Hosie, R. C. Native trees of Canada. Eighth ed., Fitzhenry and Whiteside Pubs. Ltd., Don Mills, Ontario. 64; 1979.
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.
Lu, C-Y.; Thorpe, T. A. Somatic embryogenesis and plantlet regeneration in cultured immature embryos ofPicea glauca. J. Plant Physiol. 128:297–302; 1987.
Maheshwari, P. An introduction to the embryology of angiosperms. New York, McGraw Hill; 1950.
McCready, R. M.; Guggolz, J.; Silviera, V., et al. Determination of starch and amylase in vegetables. Anal. Chem. 22:1156–1158; 1950.
Misra, S.; Green, M. J. Developmental gene expression in conifer embryogenesis and germination. I. Seed proteins and protein body composition of mature embryo and the megagametophyte of white spruce (Picea glauca [Moench] voss.). Plant Sci. 68:163–173; 1990.
Owens, J. N.; Molder, M. The reproductive cycle of Interior spruce. Province of British Columbia, Ministry of Forests, Information Services Branch, Victoria. 1984.
Reynolds, E. S. The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17:208–212; 1963.
Roberts, D. R.; Flinn, B. S.; Webb, D. T., et al. Abscisic acid and indole-3-butyric acid regulation of maturation and accumulation of storage proteins in somatic embryos of interior spruce. Physiol. Plant. 78:355–360; 1990.
Roberts, D. R.; Flinn, B. S.; Webb, D. T. Characterization of immature embryos of interior spruce by SDS-PAGE and microscopy in relation to their competence for somatic embryogenesis. Plant Cell Rep. 8:285–288; 1989.
Singh, H. Embryology of gymnosperms. Encyclopedia of Plant Anatomy, vol 0, pt. 2. Gerbruder Borntraeger, Berlin; 1978.
Spector, T. Refinement of the coomassie blue method of protein quantitation. Anal. Biochem. 86:142–146; 1978.
Sussex, I.; Clutter, M.; Walbot, V., et al. Biosynthetic activity of the suspensor ofPhaseolus coccineus. Caryologia 25 Suppl., 261–272; 1973.
von Arnold, S.; Eriksson, T. In vitro studies of adventitious shoot formation inPinus contorta. Can. J. Bot. 59:870–874; 1981.
von Arnold, S.; Hakman, I. Regulation of somatic embryo development inPicea abies by abscisic acid (ABA). J. Plant Physiol. 132:164–169; 1988.
Walbot, V.; Brady, T.; Clutter, M., et al. Macromolecular synthesis during plant embryogeny: Rates of RNA synthesis inPhaseolus coccineus embryos and suspensors. Develop. Biol. 29:104–111; 1972.
Walthall, E. D.; Brady, T. The effect of the suspensor and gibberellic acid ofPhaseolus vulgaris embryo protein synthesis. Cell Differ. 18:37–44; 1986.
Yeung, E. C. Embryogeny ofPhaseolus: the role of the suspensor. Z. Pflanzenphysiol. 96:17–28; 1980.
Yeung, E. C. Histological and histochemical staining procedures. In: Vasil, I. K. ed. Cell culture and somatic cell genetics of plants. Orlando, FL: Academic Press; 1984:689–697.
Yeung, E. C. A simple procedure to visualize osmicated storage lipids in semithin epoxy sections of plant tissues. Stain Technol. 65:45–47; 1990.
Yeung, E. C.; Law, S. Serial sectioning techniques for a modified LKB historesin. Stain Technology 62:147–153; 1987.
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Joy, R.W., Yeung, E.C., Kong, L. et al. Development of white spruce somatic embryos: I. Storage product deposition. In Vitro Cell Dev Biol - Plant 27, 32–41 (1991). https://doi.org/10.1007/BF02632059
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DOI: https://doi.org/10.1007/BF02632059