Abstract
Total protein patterns were studied in the course of development of pea somatic embryos using simple protocol of direct regeneration from shoot apical meristems on auxin supplemented medium. Protein content and total protein spectra (SDS-PAGE) of somatic embryos in particular developmental stages were analysed in Pisum sativum, P. arvense, P. elatius and P. jomardi. Expression of seed storage proteins in somatic embryos was compared with their accumulation in zygotic embryos of selected developmental stages. Pea vegetative tissues, namely leaf and root, were used as a negative control not expressing typical seed storage proteins. The biosynthesis and accumulation of seed storage proteins was observed during somatic embryo development (since globular stage), despite of the fact that no special maturation treatment was applied. Major storage proteins typical for pea seed (globulins legumin, vicilin, convicilin and their subunits) were detected in somatic embryos. In general, the biosynthesis of storage proteins in somatic embryos was lower as compared to mature dry seed. However, in some cases the cotyledonary somatic embryos exhibited comparatively high expression of vicilin, convicilin and pea seed lectin, which was even higher than those in immature but morphologically fully developed zygotic embryos. Desiccation treatments did not affect the protein content of somatic embryos. The transfer of desiccated somatic embryos on hormone-free germination medium led to progressive storage protein degradation. The expression of true seed storage proteins may serve as an explicit marker of somatic embryogenesis pathway of regeneration as well as a measure of maturation degree of somatic embryos in pea.
Similar content being viewed by others
Abbreviations
- MSB-medium:
-
medium with mineral salts after Murashige and Skoog (1962) and vitamins after Gamborg et al. (1968)
- SDS-PAGE:
-
sodium dodecylsulphate polyacrylamide gel electrophoresis
- SE:
-
somatic embryo
- ZE:
-
zygotic embryo
References
Bradford, M.M.: A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.-Anal. Biochem. 72: 248–254, 1976.
Casey, R.: Lipoxygenases.-In: Shewry, P.R., Casey, R. (ed.): Seed Proteins. Pp. 685–708. Kluwer Acad. Publ., Dordrecht 1999.
Casey, R., Domoney, C.: Pea globulins.-In: Shewry, P.R., Casey, R. (ed.): Seed Proteins. Pp. 171–208. Kluwer Acad. Publ., Dordrecht 1999.
Casey, R., Domoney, C., Ellis, N.: Legume storage proteins and their genes.-In: Miflin, B.J. (ed.): Oxford Surveys of Plant Molecular and Cell Biology. Vol. 3. Pp. 1–95. Oxford University Press, Oxford-New York 1986.
Casey, R., Domoney, C., Smith, A.M.: Biochemistry and molecular biology of seed products.-In: Casey, R., Davies, D.R. (ed.): Peas: Genetics, Molecular Biology and Biotechnology. Pp. 121–163. CAB International, Wallingford 1993.
Chanprame, S., Kuo, T.M., Widholm, J.M.: Soluble carbohydrate content of soybean [Glycine max (L.) Merr.] somatic and zygotic embryos during development.-In Vitro cell. dev. Biol. Plant 34: 64–68, 1998.
Chanprame, S., Widholm, J.M.: Comparison of oil, protein and sugars content in soybean (Glycine max) cv. Jack somatic and zygotic embryos.-In Vitro cell. dev. Biol. 32: 90A, 1996.
Christou, P., Yang, N.-S.: Developmental aspects of soybean (Glycine max) somatic embryogenesis.-Ann. Bot. 64: 225–234, 1989.
Dahmer, M.L., Hildebrand, D.F., Collins, G.B.: Comparative protein accumulation patterns in soybean somatic and zygotic embryos.-In Vitro cell. dev. Biol. 28P: 106–114, 1992.
Gamborg, O.L., Miller, R.A., Ojima, K.: Nutrient requirements of suspension cultures of soybean root cells.-Exp. Cell Res. 50: 151–158, 1968.
Griga, M.: The study of in vitro regeneration systems in pea, horse bean and soybean.-Ph.D. Thesis, Masaryk University, Brno 1990. [In Czech.]
Griga, M.: Direct somatic embryogenesis from shoot apical meristems of pea and thidiazuron induced high conversion rate of somatic embryos.-Biol. Plant. 41: 481–495, 1998.
Griga, M.: Somatic embryogenesis in grain legumes.-In: Strnad, M., Peč, P. (ed.): Advances in Regulation of Plant Growth and Development. Pp. 233–249. Peres Publ., Prague 1999.
Griga, M.: Morphology and anatomy of Pisum sativum somatic embryos.-Biol. Plant. 45: 173–182, 2002.
Griga, M., Horáček, J., Klenotičová, H.: Biochemical description of pea somatic embryogenesis.-In: Abstr. 10th IAPTCB Congress — Plant Biotechnology 2002 and Beyond. Pp. 131–A, P-1392. IAPTCB Publ., Orlando 2002.
Griga, M., Pošvec, Z., Horáček, J., Klenotičová, H: Protein and starch metabolism during development of pea somatic embryos.-In: Proc. 4th Europ. Conf. Grain Legumes — Towards The Sustainable Production of Healthy Food, Feed and Novel Products. Pp. 362–363. AEP Publ., Paris 2001.
Griga, M., Stejskal, J.: Auxin and sucrose effects on maturation of soybean somatic embryos from long-term repetitive culture.-Biol. Plant. 36(Suppl.): 81, 1994.
Jacobsen, H.-J.: Somatic embryogenesis in seed legumes: the possible role of soluble auxin receptors.-Israel J. Bot. 40: 139–143, 1991.
Jacobsen, H.-J., Kysely, W.: Induction of somatic embryos in pea, Pisum sativum L.-Plant Cell Tissue Organ Cult. 3: 319–324, 1994.
Komatsuda, T., Lee, W., Oka, S.: Maturation and germination of somatic embryos as affected by sucrose and plant growth regulators in soybeans Glycine gracilis Skwortz and Glycine max (L.) Merr.-Plant Cell Tissue Organ Cult. 28: 103–113, 1992.
Krochko, J.E., Bantroch, D.J., Greenwood, J.S., Bewley, J.D.: Seed storage proteins in developing somatic embryos of alfalfa: defects in accumulation compared to zygotic embryos.-J. exp. Bot. 45: 699–708, 1994.
Krochko, J.E., Pramanik, S.K., Bewley, J.D.: Contrasting storage protein synthesis and messenger RNA accumulation during development of zygotic and somatic embryos of alfalfa (Medicago sativa L.).-Plant Physiol. 99: 46–53, 1992.
Kysely, W., Jacobsen, H.-J.: Somatic embryogenesis from pea embryos and shoot apices.-Plant Cell Tissue Organ Cult. 20: 7–14, 1990.
Kysely, W., Myers, J.R., Lazzeri, P.A., Collins, G.B., Jacobsen, H.-J.: Plant regeneration via somatic embryogenesis in pea (Pisum sativum L.).-Plant Cell Rep. 6: 305–308, 1987.
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4.-Nature 227: 680–685, 1970.
Lai, F.-M., McKersie, B.D.: Regulation of starch and protein accumulation in alfalfa (Medicago sativa L.) somatic embryos.-Plant Sci. 100: 211–219, 1994a.
Lai, F.-M., McKersie, B.D.: Regulation of storage protein synthesis by nitrogen and sulfur nutrients in alfalfa (Medicago sativa L.) somatic embryos.-Plant Sci. 103: 209–221, 1994b.
Lai, F.-M., Senaratna, T., McKersie, B.D.: Glutamine enhances storage protein synthesis in Medicago sativa L. somatic embryos.-Plant Sci. 87: 69–77, 1992.
Lecouteux, C.G., Lai, F.-M., McKersie, B.D.: Maturation of alfalfa (Medicago sativa L.) somatic embryos by abscisic acid, sucrose and chilling stress.-Plant Sci. 94: 207–213, 1993.
Lehminger-Mertens R., Stirn, S., Jacobsen, H.-J.: Storage protein patterns of somatic and zygotic pea embryos.-In: Abstracts VIIth Int. Congr. Plant Tissue Cell Cult. Pp. 257. IAPTC, Amsterdam 1990.
Liu, C.M., Johnson, S., Hedley, C.L., Wang, T.L.: The generation of a legume embryo: morphological and cellular defects in pea mutants.-In: Wang, T.L., Cuming, A.C. (ed): Embryogenesis, the Generation of a Plant. Pp. 191–213. Bios Scientific Publishers, Oxford 1996.
Liu, C.M., Johnson, S., Wang, T.L.: cyd, a mutant of pea that alters embryo morphology is defective in cytokinesis.-Dev. Genet. 16: 321–331, 1995.
Loiseau, J., Michaux-Ferriere, N., Le Deunff, Y.: Histology of somatic embryogenesis in pea.-Plant Physiol. Biochem. 36: 683–687, 1998.
Marinos, N.G.: Embryogenesis of the pea (Pisum sativum). I. The cytological environment of the developing embryo.-Protoplasma 70: 261–279, 1970.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassay with tobacco tissue cultures.-Physiol. Plant. 15: 472–497, 1962.
Pate, J.S.: Pea.-In: Evans, L.T. (ed.): Crop Physiology. Pp. 173–196. Cambridge Univ. Press, Cambridge 1975.
Peumans, W.J., Van Damme, E.J.M.: Seed lectins.-In: Shewry, P.R., Casey, R. (ed.): Seed Proteins. Pp. 657–683. Kluwer Acad. Publ., Dordrecht 1999.
Shewry, P.R., Casey, R.: Seed proteins.-In: Shewry, P.R., Casey, R. (ed.): Seed Proteins. Pp. 1–10. Kluwer Acad. Publ., Dordrecht 1999.
Slawinska, J., Obendorf, R.L.: Soybean somatic embryo maturation: composition, respiration and water relations.-Seed Sci. Res. 1: 251–262, 1991.
Stejskal, J.: Comparative analysis of proteins and isozymes in somatic and zygotic embryos of soybean (Glycine max (L.) Merr.).-Ph. D. Thesis, IEB Prague 1994. [In Czech.]
Stejskal, J., Griga, M.: Comparative analysis of some isozymes and proteins in somatic and zygotic embryos of soybean (Glycine max (L.) Merr.).-J. Plant Physiol. 146: 497–502, 1995.
Stirn, S., Jacobsen, H.-J.: Marker proteins for embryogenic differentiation patterns in pea callus.-Plant Cell Rep. 6: 50–54, 1987.
Stirn, S., Jacobsen, H.-J.: Production and characterization of monoclonal antibodies against marker proteins for somatic embryogenesis in pea (Pisum sativum L.).-In: Nijkamp, H.J.J., Van Der Plas L.H.W., Van Aartrijk J. (ed.): Progress in Plant Cellular and Molecular Biology. Pp. 460–465. Kluwer Acad. Publ., Amsterdam 1990.
Stuart, D.A., Nelsen, J., Nichol, J.W.: Expression of 7S and 11S alfalfa seed storage proteins in somatic embryos.-J. Plant Physiol. 132: 134–139, 1988.
Vitale, A., Bollini, R.: Legume storage proteins.-In: Kigel, J., Galili, G. (ed.): Seed Development and Germination. Pp. 73–102. Marcel Dekker Inc., New York 1995.
Wang, T.L., Hedley, C.L.: Genetic and developmental analysis of the seed.-In: Casey, R., Davies, D.R. (ed.): Peas: Genetics, Molecular Biology and Biotechnology. Pp. 83–120. CAB International, Wallingford 1993.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Griga, M., Horáček, J. & Klenotičová, H. Protein patterns associated with Pisum sativum somatic embryogenesis. Biol Plant 51, 201–211 (2007). https://doi.org/10.1007/s10535-007-0041-9
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10535-007-0041-9