Skip to main content
Log in

The initiation of legumin synthesis in immature embryos of Pisum sativum L. grown in vivo and in vitro

  • Published:
Planta Aims and scope Submit manuscript

Abstract

A highly sensitive immunoassay has been used for the detection of a major storage protein, legumin, in embryos of Pisum sativum L.; with this technique nanogram quantities could be measured. In the two varieties tested, legumin could be detected in embryos in vivo, when they had attained a fresh weight of 2·10-3 g and 3·10-3 g, respectively. Contrary to earlier claims, embryos cultured in vitro were shown to be capable of initiating legumin synthesis. This capacity to initiate legumin synthesis was confirmed by two-dimensional isoelectric focusing-electrophoresis and fluorography; embryos harvested before initiation of legumin synthesis and cultured in radioactive medium were shown to have synthesized legumin subunits. The amounts of legumin and total protein synthesized per unit fresh weight were consistently greater in vitro than in equivalent embryos grown in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

ELISA:

Enzyme-linked immunosorbent assay

BSA:

bovine serum albumin

IgG:

immunoglobulin

SDS:

sodium dodecyl sulphate

DSP:

Pisum cv. Dark Skinned Perfection

References

  • Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88

    PubMed  Google Scholar 

  • Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254

    Article  PubMed  Google Scholar 

  • Carasco, J.F., Croy, R., Derbyshire, E., Boulter, D. (1978) The isolation and characterization of the major polypeptides of the seed globulin of cowpea (Vigna unguiculata L. Walp) and their sequential synthesis in developing seeds. J. Exp. Bot. 29, 309–323

    Google Scholar 

  • Casey, R (1979) Immunoaffinity chromatography as a means of purifying legumin from Pisum (pea) seeds. Biochem. J. 177, 509–520

    PubMed  Google Scholar 

  • Clark, M.F., Adams, A.N. (1977) Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol. 34, 475–483

    PubMed  Google Scholar 

  • Cullis, C.A. (1978) Chromatin-bound DNA-dependent RNA polymerase in developing pea cotyledons. Planta 144, 57–62

    Google Scholar 

  • Davies, D.R. (1976) Variation in the storage proteins of peas in relation to sulphur amino-acid content. Euphytica 25, 717–724

    Google Scholar 

  • Dudman, W.F., Millerd, A. (1975) Immunochemical behaviour of legumin and vicilin from Vicia faba: a survey of related proteins in the Leguminosae subfamily Faboideae. Biochem. Syst. Ecol. 3, 25–33

    Google Scholar 

  • Dure, L.S. (1975) Seed formation. Annu. Rev. Plant Physiol. 26, 259–278

    Article  Google Scholar 

  • Guldager, P. (1978) Immunoelectrophoretic analysis of seed proteins from Pisum sativum L. Theor. Appl. Genet. 53, 241–250

    Google Scholar 

  • Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685

    Google Scholar 

  • Laskey, R.A., Mills, A.D. (1975) Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur. J. Biochem. 56, 335–341

    PubMed  Google Scholar 

  • Lea, P.J., Hughes, J.S., Miflin, B.J. (1979) Glutamine- and asparagine-dependent protein synthesis in maturing legume cotyledons cultured in vitro. J. Exp. Bot. 30, 529–537

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. (1951) Protein measurement with Folin phenol reagent. J. Biol. Chem. 193, 265–275

    PubMed  Google Scholar 

  • Millerd, A., Spencer, D. (1974) Changes in RNA-synthesizing activity and template activity in nuclei from cotyledons of developing pea seeds. Aust. J. Plant Physiol. 1, 331–341

    Google Scholar 

  • Millerd, A., Spencer, D., Dudman, W.F., Stiller, M. (1975) Growth of immature pea cotyledons in culture. Aust. J. Plant Physiol. 2, 51–59

    Google Scholar 

  • Millerd, A., Thomson, J.A., Schroeder, H.E. (1978) Cotyledonary storage proteins in Pisum sativum. III. Patterns of accumulation during development. Aust. J. Plant Physiol. 5, 519–534

    Google Scholar 

  • O'Farrell, P.H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021

    PubMed  Google Scholar 

  • Scholz, G., Richter, J., Manteuffel, R. (1974) Studies on seed globulins from legumes. I. Separation and purification of legumin and vicilin from Vicia faba L. by zone precipitation. Biochem. Physiol. Pflanzen 166, 163–172

    Google Scholar 

  • Stafford, A., Davies, D.R. (1979) The culture of immature pea embryos. Ann. Bot. 44, 315–321

    Google Scholar 

  • Thompson, J.F., Madison, J.T., Muenster, A.E. (1977) In vitro culture of immature cotyledons of soya bean (Glycine max L. Merr.) Ann. Bot. 41, 29–39

    Google Scholar 

  • Voller, A., Bartlett, A., Bedwell, D.E., Clark, M.F., Adams, A.N. (1976) The detection of viruses by enzyme-linked immunosorbent assay (ELISA). J. Gen. Virol. 33, 165–167

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Domoney, C., Davies, D.R. & Casey, R. The initiation of legumin synthesis in immature embryos of Pisum sativum L. grown in vivo and in vitro. Planta 149, 454–460 (1980). https://doi.org/10.1007/BF00385747

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00385747

Key words

Navigation