Hostname: page-component-848d4c4894-5nwft Total loading time: 0 Render date: 2024-06-02T01:19:46.063Z Has data issue: false hasContentIssue false

Haemoglobin expression in germinating barley

Published online by Cambridge University Press:  19 September 2008

Stephen M.G. Duff
Affiliation:
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
Phillip A. Guy
Affiliation:
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
Xianzhou Nie
Affiliation:
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
Douglas C. Durnin
Affiliation:
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
Robert D. Hill*
Affiliation:
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
*
*Tel: 204-474-6087 Fax: 204-474-7525 E-mail: rob_hill/@cc.umanitoba.ca

Abstract

Polyclonal antibodies to purified recombinant barley haemoglobin (Hb) have been raised in rabbits and used to investigate its expression in monocotyledonous plants. Very little or no Hb expression was observed in dry barley seeds but germination resulted in the expression of Hb which peaked at 2–3 days after imbibition. Hb expression was also observed in maize, wheat, wild oat and Echinochloa crus-galli seeds during germination. Dissection of tissues from the barley seedlings showed that most of the haemoglobin was expressed in the root and seed coat (aleurone layer), with very little in the coleoptile. Imbibition of half-seeds or excised embryos resulted in the expression of haemoglobin. ATP measurements of barley embryos showed that ATP levels quickly increase after imbibition. α-Amylase activity was also determined in embryos to correlate Hb expression with a well-characterized germination response. The results demonstrate that Hb expression is a normal consequence of germination.

Type
Physiology & Biochemistry
Copyright
Copyright © Cambridge University Press 1998

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Andersson, C.R., Jensen, E.O., Llewellyn, D.J., Dennis, E.S. and Peacock, W.J. (1996) A new hemoglobin gene from soybean, A role for hemoglobin in all plants. Proceedings of the National Academy of Sciences, USA 93, 427431.CrossRefGoogle Scholar
Appleby, C.A. (1992) The origin and function of haemoglobin in plants. Science Progress 76, 365398.Google Scholar
Appleby, C.A., Tjepkema, J.D. and Trinick, M.J. (1983) Hemoglobin in a nonleguminous plant, Parasponia: Possible genetic origin and function in nitrogen fixation. Science 220, 951954.Google Scholar
Arredondo-Peter, R., Hargrove, M.S., Sarath, G., Moran, J.F., Lohrman, J., Olson, J.S. and Klucas, R.V. (1997) Rice hemoglobins: Gene cloning, analysis and oxygen binding kinetics of a recombinant protein synthesized in Escherichia coli. Plant Physiology 115, 12591266.CrossRefGoogle ScholarPubMed
Bewley, J.D. and Black, M. (1994) Cellular events during germination and seedling growth. pp 147197 in Seeds. Physiology of development and germination. New York, Plenum Press.Google Scholar
Bogusz, D., Appleby, C.A., Landsmann, J., Dennis, E.S., Trinick, M.J. and Peacock, W.J. (1988) Functioning haemoglobin in non-nodulating plants. Nature 131, 178180.CrossRefGoogle 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. Analytical Biochemistry 72, 248254.Google Scholar
Briggs, D.E. (1961) A modification of the Sandstedt, Kneen and Blish assay of α-amylase. Journal of the Institute of Brewing 67, 427431.Google Scholar
Brown, G.G., Lee, J.S., Brisson, N. and Verma, D.P. (1984) The evolution of a plant globin gene family. Journal of Molecular Evolution 21, 1932.Google Scholar
Christensen, T., Dennis, F.S., Peacock, J.W., Landsmann, J. and Marker, K.A. (1991) Haemoglobin genes in nonlegumes: cloning and characterisation of a Casuarina glauca haemoglobin gene. Plant Molecular Biology 16, 339344.Google Scholar
Duff, S.M.G., Plaxton, W.C. and Lefebvre, D.D. (1991) Phosphate-starvation response in plant cells: De novo synthesis and degradation. Proceedings of the National Academy of Sciences, USA 88, 95389542.Google Scholar
Duff, S.M.G., Wittenberg, J.B. and Hill, R.D. (1997) Expression, purification and properties of recombinant barley (Hordeum sp.) Hemoglobin. Optical spectra and reactions with gaseous ligands. Journal of Biological Chemistry 272, 1674616752.Google Scholar
Hardison, R.C. (1996) A brief history of hemoglobins: Plant, animal, protist and bacteria. Proceedings of the National Academy of Sciences, USA 93, 56755682.Google Scholar
Johnston, F.B. and Stern, H. (1957) Mass isolation of viable wheat embryos. Nature 179, 160161.CrossRefGoogle ScholarPubMed
Keilin, F.R.S. and Wang, Y.L. (1945) Hemoglobin in the root nodules of leguminous plants. Nature 155, 227229.CrossRefGoogle Scholar
Kubo, H. (1939) Über Hämoprotein aus den Wurzelknöllchen von Leguminosin. Acta Phytochimica 11, 195200.Google Scholar
Landsmann, J., Dennis, E.S., Higgins, T.J.V., Appleby, C.A., Kortt, A.A. and Peacock, W.J. (1986) Common evolutionary origin of legume and non-legume plant haemoglobins. Nature 324, 166168.Google Scholar
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.Google Scholar
Lin, M., Turpin, D.T. and Plaxton, W.C. (1989) Pyruvate kinase isozymes from the green algae Selanstrum minutum I. Purification, physical and immunological characterization. Archives of Biochemistry and Biophysics 269, 219227.Google Scholar
Lowry, O.H. and Passonneau, J.V. (1972) A collection of metabolite assays. pp 146222in A flexible system of enzymatic analysis. Academic Press, New York and London.Google Scholar
Nie, X-Z. and Hill, R.D. (1997) Mitochondrial respiration and hemoglobin gene expression in barley aleurone tissue. Plant Physiology 114, 835840.CrossRefGoogle ScholarPubMed
Osborne, D.J. (1983) Biochemical control systems operating in the early hours of germination. Canadian Journal of Botany 61, 35683577.CrossRefGoogle Scholar
Plaxton, W.C. (1989) Molecular and immunological characterization of plastid and cytosolic pyruvate kinase isozymes from castor-oil-plant endosperm and leaf. European Journal of Biochemistry 181, 443451.Google Scholar
Sowa, A., Duff, S.M.G., Guy, P.A. and Hill, R.D. (1998) Altering hemoglobin levels changes energy status in maize cells under hypoxia. Proceedings of the National Academy of Sciences, USA 95, 1031710321.CrossRefGoogle ScholarPubMed
Taylor, E.R., Nie, X-Z., MacGregor, A.W. and Hill, R.D. (1994) A cereal haemoglobin gene is expressed in seed and root tissues under anaerobic conditions. Plant Molecular Biology 24, 853862.Google Scholar
Trevaskis, B., Watts, R.A., Andersson, C.R., Llewellyn, D.J., Hargrove, M.S., Olson, J.S., Dennis, E.S. and Peacock, W.J. (1997) Two hemoglobin genes in Arabidopsis thaliana: The evolutionary origins of leghemoglobins. Proceedings of the National Academy of Sciences, USA 94, 1223012234.Google Scholar