Abstract
The ability of soybean (Glycine max (L.) Merr.) embryos cultured in vitro in the absence of abscisic acid (ABA) to germinate precociously increased as the embryos matured. Exogenous ABA prevented precocious germination at all stages of development, concentrations below 10-5 M being partially effective. Growth (fresh weight increase) of mid-maturation embryos was dependent on ABA, and such embryos required ABA for continued synthesis of storage proteins during culture. Two complementary-DNA clones for different members of the family of 11S storage proteins, and one for the 7S storage proteins were used in Northern blot hybridizations to analyze the effects of ABA on the levels of the 11S and 7S mRNAs. In addition, filter hybridizations with in-vivo-labeled [3H]polyadenylated RNA to the cloned DNA for one of the 11S proteins were carried out to study transcription of 11S mRNA. Midmaturation embryos cultured with ABA continued to transcribe mRNAs for the storage proteins during the 21 d of culture studied, whereas in the absence of ABA these mRNAs disappeared from the cotyledons within about 5 d. The optimum concentration of ABA for the synthesis of storage-protein mRNAs was 10-5 M. The effect of ABA on the concentration of 11S mRNAs was stage-dependent. Abscisic acid caused a decrease in the mRNA in embryos cultured at the cotyledon stage; it was necessary for high levels of the mRNA to be achieved in early- and mid-maturation embryos; and it did not reverse the decline in the mRNA levels in embryos cultured at the late-maturation stage.
Similar content being viewed by others
Abbreviations
- ABA:
-
abscisic acid
- Da:
-
dalton
- FW:
-
fresh weight
- Poly(A)RNA:
-
polyadenylated RNA
- SDS:
-
sodium dodecyl sulfate
References
Ackerson, R.C. (1984a) Regulation of soybean embryogenesis by abscisic acid. J. Expt. Bot. 35, 403–413
Ackerson, R.C. (1984b) Abscisic acid and precocious germination in soybeans. J. Expt. Bot. 35, 414–421
Altschuler, M., Mascarenhas, J.P. (1982) Heat shock proteins and effects of heat shock in plants. Plant Mol. Biol. 1, 103–115
Bingham, P.M., Judd, B.H. (1981) A copy of the copia transposable element is very tightly linked to the w a allele at the white locus of Drosophila melanogaster. Cell 25, 705–711
Choinski, J.S., Trelease, R.N. (1978) Control of enzyme activities in cotton cotyledons during maturation and germination. II. Glyoxysomal enzyme development in embryos. Plant Physiol. 62, 141–145
Clewell, D.B., Helinski, D.R. (1972) Effect of growth conditions on the formation of the relaxation complex of supercoiled ColEl deoxyribonucleic acid and protein in E. coli. J. Bacteriol. 110, 1135–1142
Crouch, M.L., Sussex, I.M. (1981) Development and storage protein synthesis in Brassica napus L. embryos in-vivo and in-vitro. Planta 153, 64–74
Davies, D.R., Bedford, I.D. (1982) Abscisic acid and storage protein accumulation in Pisum sativum embryos grown in in-vitro. Plant Sci. Lett. 27, 337–343
Dure, L.S., Galau, G.A. (1981) Developmental biochemistry of cottonseed embryogenesis and germination. XI. Regulation of biosynthesis of principal storage proteins. Plant Physiol. 68, 187–194
Dure, L.S., Galau, G.A., Greenway, S.C. (1980) Changing protein patterns during cotton cotyledon embryogenesis and germination shown by in-vivo and in-vitro synthesis. Israel J. Bot. 28, 142–153
Dure, L.S., Greenway, S.C., Galau, G.A. (1981) Developmental biochemistry of cottonseed embryogenesis and germination. XIV. Changing messenger RNA populations as shown by in-vivo protein synthesis. Biochemistry 20, 4162–4168
Eeuwens, C.J., Schwabe, W.W. (1975) Seed and pod wall development in Pisum sativum L. in relation to extracted and applied hormones. J. Exp. Bot. 26, 1–14
Eisenberg, A.J. (1984) The role of abscisic acid in soybean embryo development. Ph.D. thesis, State University of New York, Albany, USA.
Eisenberg, A., Altschuler, M., Haskell, D.W., Bashe, D.M., Mascarenhas, J.P. (1984) A simple and inexpensive procedure for preserving plant tissues for RNA analysis. Plant Mol. Biol. Rep. 2, 16–23
Eisenberg, A.J., Holmes, D.S. (1982) A note on the use of CsCl centrifugation to purify plasmids prepared by the rapid boiling method. Anal. Biochem. 127, 434
Finkelstein, R.R., Crouch, M.L. (1984) Precociously germinating rape seed embryos retain characteristics of embryogeny. Planta 162, 125–131
Fisher, R.L., Goldberg, R.B. (1982) Structure and flanking regions of soybean seed protein genes. Cell 29, 651–660
Goldberg, D.A. (1980) Isolation and partial characterization of the Drosophila alcohol dehydrogenase gene. Proc. Natl. Acad. Sci. USA 77, 5794–5801
Goldberg, R.B., Hoschek, G., Tam, S.H., Ditta, G.S., Breidenbach, R.W. (1981a) Abundance, diversity, and regulation of mRNA sequence sets in soybean embryogenesis. Dev. Biol. 83, 201–217
Goldberg, R.B., Hoschek, G., Ditta, G.S., Breidenbach, R.W. (1981b) Developmental regulation of cloned superabundant embryo mRNAs in soybean. Dev. Biol. 83, 218–231
Hill, J.E., Breidenbach, R.W. (1974) Proteins of soybean seeds. II. Accumulation of the major protein components during seed development and maturation. Plant Physiol. 53, 727–751
Holmes, D.S., Quigley, M. (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal. Biochem. 114, 193–197
Hsu, F.C. (1979) Abscisic acid accumulation in developing seeds of Phaseolus vulgaris L. Plant Physiol 63, 552–556
Jackson, G.A.D. (1968) Hormonal control of fruit development, seed dormancy and germination with particular reference to Rosa. Soc. Chem. Ind. (Lond.). Monogr. 31, 127–156
Kessler, S.W. (1975) Rapid isolation of antigens from cells with a staphylococcal protein A-antibody absorbant: parameters of the interaction of antibody complexes with protein A. J. Immunol. 115, 1617–1624
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
Larkins, B.A., Jones, R.A., Tsai, C.Y. (1976) Isolation and in-vitro translation of zein mRNA. Biochemistry 15, 5506–5510
Lehrach, H., Diamond, D., Wozney, J.M., Boedtker, H. (1977) RNA molecular weight determinations by gel electrophoresis under denaturing conditions: A critical re-examination. Biochemistry 16, 4743–4754
Machlis, L., Torrey, J.G. (1956) Plants in action. W.H. Freeman, San Francisco
Maniatis, T., Jeffrey, D.G., Kleid, D.G. (1975) Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. USA 72, 1184–1188
Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor
Meinke, D.W., Chen, J., Beachy, R.N. (1981) Expression of storage protein genes during soybean seed development. Planta 153, 130–139
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Obendorf, R.L., Wettlaufer, S.H. (1984) Precocious germination during in-vitro growth of soybean seeds. Plant Physiol. 76, 1024–1028
Paterson, B.W., Roberts, B.E. (1981) Structural gene identification utilizing eukaryotic cell-free translational systems. In: Gene amplification and analysis, vol. 2, pp. 417–437, Chirikjian, J.G., Papas, T.S., eds. Elsevier/North Holland, Amsterdam
Quatrano, R.S., Ballo, B.L., Williamson, J.D., Hamblin, M.T., Mansfield, M. (1983) ABA controlled expression of embryo specific genes during wheat grain development. In: UCLA Symp. Mol. Cell. Biol., N.S. vol. 12; Plant molecular biology, pp. 343–353, Goldberg, R.B., ed. Allan R. Liss, New York
Quebedeaux, B., Sweetser, P.B., Rowell, J.C. (1976) Abscisic acid levels in soybean reproductive structures during development. Plant Physiol. 58, 363–366
Sengupta, G.V., Deluca, V., Bailey, D. Verma, D.P.S. (1981) Biosynthesis of soybean storage proteins: post-translational processing of 7S and 11S components. Plant Mol. Biol. 1, 19–34
Sondheimer, E., Tzou, D.S., Galson, E.C. (1968) Abscisic acid levels and seed dormancy. Plant Physiol 43, 1443–1447
Sussex, I.M., Dale, R.M.K. (1979) Hormonal control of storage protein synthesis in Phaseolus vulgaris. In: The plant seed: development, preservation and germination, pp. 129–141, Rubenstein, I., Phillips, R.L., Green, C.E., Gengenbach, B.G., eds. Academic Press, New York London
Sussex, I.M., Dale, R.M.K., Crouch, M.L. (1980) Developmental regulation of storage protein synthesis in seeds. In: Genome organization and expression in plants, pp. 283–289, Leaver, C.J., ed. Plenum Press, New York
Thomas, P.S. (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77, 5201–5205
Young, E.T., Mattson, T., Selzer, G., Houwe, G.V., Bolle, A., Epstein, R. (1980) Bacteriophage T4 gene transcription studied by hybridization to cloned restriction fragments. J. Mol. Biol. 138, 423–445
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Eisenberg, A.J., Mascarenhas, J.P. Abscisic acid and the regulation of synthesis of specific seed proteins and their messenger RNAs during culture of soybean embryos. Planta 166, 505–514 (1985). https://doi.org/10.1007/BF00391275
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00391275