Skip to main content
SpringerLink
Log in

The occurrence of leghemoglobin protein in the uninfected interstitial cells of soybean root nodules

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The distribution of leghemoglobin (Lb) in resin-embedded root nodules of soybean (Glycine max (L.) Merr.) was investigated using immunogold labeling. Using anti-Lb immunoglobulin G and protein A-gold, Lb or its apoprotein was detected both in cells infected by Bradyrhizobium japonicum and in uninfected interstitial cells. Leghemoglobin was present in the cytoplasm, exclusive of the organelles, and in the nuclei of both cell types. In a comparison of the density of labeling in adjacent pairs of infected and uninfected cells, Lb was found to be about four times more concentrated in infected cells. This is the first report of Lb in uninfected cells of any legume nodule; it raises the possibility that this important nodule-specific protein may participate in mediating oxygen flow to host plant organelles throughout the infected region of the nodule.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

IgG:

immunoglobulin G

kDA:

kilodalton

Lb:

leghemoglobin

TBST:

Tris-buffered saline plus Tween 20

References

  • Appleby, C.A. (1984) Leghemoglobin and Rhizobium respiration. Annu. Rev. Plant Physiol. 35, 443–478

    Google Scholar 

  • Appleby, C.A. (1985) Plant hemoglobin properties, function and genetic origin. In: Nitrogen fixation and CO2 metabolism, pp. 41–51. Ludden, P.W., Burris, J.E., eds. Elsevier Science Publishing Co., New York Amsterdam Oxford

    Google Scholar 

  • Appleby, C.A., Tjepkema, J.D., Trinick, M.J. (1983) Hemoglobin in a non-leguminous plant, Parasponia: possible genetic origin and function in nitrogen fixation. Science 220, 951–953

    Google Scholar 

  • Bergerson, F.J., Appleby, C.A. (1981) Leghaemoglobin within bacteroid-enclosing membrane envelopes from soybean root nodules. Planta 152, 534–543

    Google Scholar 

  • Bergerson, F.J., Turner, G.L., Appleby, C.A. (1973) Studies on the physiological role of leghemoglobin in soybean root nodules. Biochim. Biophys. Acta 292, 271–282

    Google Scholar 

  • Bergmann, H., Preddie, E., Berma, D.P.S. (1983) Nodulin-35: a subunit of specific uricase (uricase II) induced and localized in the uninfected cells of soybean nodules. EMBO J. 2, 2333–2339

    Google Scholar 

  • Bradley, D.J., Wood, E.A., Galfre, G., Butcher, G.W., Brewin, N.J. (1988) Isolation of monoclonal antibodies reacting with peribacteroid membranes and other components of pea root nodules containing Rhizobium leguminosarum. Planta 173, 149–160

    Google Scholar 

  • Chua, N.-H., Bartlett, S.G., Weiss, M. (1982) Preparation and characterization of antibodies to chloroplast proteins. In: Methods in chloroplast molecular biology, pp. 1063–1080, Edelman, M., Hallick, R.B., Chua, N.-H., eds. Elsevier Biomedical Press, Amsterdam New York Oxford

    Google Scholar 

  • Dingwall, C., Laskey, R.A. (1986) Protein import into the nucleus. Annu. Rev. Cell Biol. 2, 367–390

    Google Scholar 

  • Doman, D.C., Trelease, R.N. (1985) Protein A-gold immunocytochemistry of isocitrate lyase in cotton seeds. Protoplasma 124, 157–167

    Google Scholar 

  • Erwee, M.G., Goodwin, P.B. (1985) Symplast domains in extrastelar tissues of Egeria densa Planch. Planta 163, 9–19

    Google Scholar 

  • Fuchsman, W.H., Appleby, C.A. (1979) Separation and determination of the relative concentrations of the homogeneous components of soybean leghemoglobin by iso-electric focusing. Biochim. Biophys. Acta 579, 314–324

    Google Scholar 

  • Govers, F., Moerman, M., Hooymans, J., van Kammen, A., Bisseling, T. (1986) Microaerobiosis is not involved in the induction of pea nodulin gene expression. Planta 169, 513–517

    Google Scholar 

  • Hanks, J.F., Schubert, K., Tolbert, N.E. (1983) Isolation and characterization of infected and uninfected cells from soybean nodules. Plant Physiol. 71, 869–873

    Google Scholar 

  • Hoagland, D.R., Snyder, W.C. (1933) Nutrition of strawberry plant under controlled conditions. Proc. Am. Soc. Hort. Sci. 30, 288–294

    Google Scholar 

  • Kyhse-Anderson, J. (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J. Biochem. Biophys. Meth. 10, 203–209

    Google Scholar 

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

    Google Scholar 

  • Lucas, K., Boland, M.J., Schubert, K.R. (1983) Uricase from soybean root nodules: purification, properties, and comparison with the enzyme from cowpea. Arch. Biochem. Biophys. 226, 190–197

    Google Scholar 

  • Newcomb, E.H., Tandon, S.R. (1981) Uninfected cells of soybean root nodules: ultrastructure suggests key role in ureide production. Science 212, 1394–1396

    Google Scholar 

  • Newcomb, E.H., Tandon, S.R., Kowal, R.R. (1985) Ultrastructural specialization for ureide production in uninfected cells of soybean root nodules. Protoplasma 125, 1–12

    Google Scholar 

  • Noel, K.D., Stacey, G., Tandon, S.R., Silver, L.E., Brill, W.J. (1982) Rhizbbium japonicum mutants defective in symbiotic nitrogen fixation. J. Bacteriol. 152, 485–494

    Google Scholar 

  • O'Brian, M.R., Kirshbom, P.M., Maier, R.J. (1987) Bacterial heme synthesis is required for expression of the leghemoglobin holoprotein but not the apoprotein in soybean root nodules. Proc. Natl. Acad. Sci. USA 84, 8390–8393

    Google Scholar 

  • Posthuma, G., Slot, J.W., Geuze, H.J. (1986) A quantitative immuno-electron microscopic study of amylase and chymotrypsinogen in peri- and tele-insular cells of the rat exocrine pancreas. J. Histochem. Cytochem. 34, 203–207

    Google Scholar 

  • Reynolds, E.S. (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–213

    Google Scholar 

  • Robertson, J.G., Warburton, M.P., Lyttleton, P., Fordyce, A.M., Bullivant, S. (1978) Membranes in lupin root nodules. II. Preparation and properties of peribacteroid membranes and bacterial envelope inner membranes from developing lupin nodules. J. Cell Sci. 30, 151–174

    Google Scholar 

  • Robertson, J.G., Wells, B., Bisseling, T., Farnden, K.J.F., Johnston, W.A.B. (1984) Immunogold localization of leghaemoglobin in cytoplasm in nitrogen-fixing root nodules of pea. Nature 311, 254–256

    Google Scholar 

  • Saari, L.L., Klucas, R.V. (1984) Ferric leghemoglobin reductase from soybean root nodules. Arch. Biochem. Biophys. 231, 102–113

    Google Scholar 

  • Selker, J.M.L., Newcomb, E.H. (1986) Spatial relationships between uninfected and infected cells in root nodules of soybean. Planta 165, 446–454

    Google Scholar 

  • Sheehy, J. E., Bergerson, F.J. (1986) A simulation study of the functional requirements and distribution of leghaemoglobin in relation to biological nitrogen fixation in legume root nodules. Ann. Bot. 58, 121–136

    Google Scholar 

  • Smith, A.M. (1985) Capacity for fermentation in roots of Rhizobium nodules of Pisum sativum L. Planta 166, 264–270

    Google Scholar 

  • Smith, J.D. (1949) The concentration and distribution of haemoglobin in the root nodules of leguminous plants. Biochem. J. 44, 585–591

    Google Scholar 

  • Tjepkema, J.D. (1983) Hemoglobins in the nitrogen-fixing root nodules of actinorhizal plants. Can. J. Bot. 61, 2924–2929

    Google Scholar 

  • Tjepkema, J.D., Yocum, C.S. (1974) Measurement of oxygen partial pressure within soybean nodules by oxygen microelectrodes. Planta 119, 351–360

    Google Scholar 

  • Tucker, E.B. (1982) Translocation in the staminal hairs of Setcreasea purpurea. 1. A study of cell ultrastructure and cellto-cell passage of molecular probes. Protoplasma 113, 193–201

    Google Scholar 

  • VandenBosch, K.A., Newcomb, E.H. (1986) Immunogold localization of nodule-specific uricase in developing soybean root nodules. Planta 167, 425–436

    Google Scholar 

  • Verma, D.P.S., Bal, A.K. (1976) Intracellular site of synthesis and localization of leghaemoglobin in soybean root nodules. Proc. Natl. Acad. Sci. USA 73, 3843–3847

    Google Scholar 

  • Verma, D.P.S., Ball, S., Guerin, C., Wanamaker, L. (1979) Leghemoglobin biosynthesis in soybean root nodules. Characterization of the nascent and released peptides and the relative rate of synthesis of the major leghemoglobins. Biochemistry 18, 476–483

    Google Scholar 

  • Weibel, E.R. (1979) Stereological methods, vol. 1. Academic Press, New York London

    Google Scholar 

  • Whittaker, R.G., Lennox, S., Appleby, C.A. (1981) Relationship of the minor soybean leghaemoglobins d1, d2 and d3 to the major leghemoglobins c1, c2, and c3. Biochem. Int. 3, 117–124

    Google Scholar 

  • Wittenberg, J.B., Bergerson, F.J., Appleby, C.A., Turner, G.L. (1974) Facilitated oxygen diffusion. The role of leghemoglobin in nitrogen fixation by bacteroids isolated from soybean root nodules. J. Biol. Chem. 249, 4057–4066

    Google Scholar 

  • Witty, J.F., Skot, L., Revsbech, N.P. (1987) Direct evidence for changes in the resistance of legume root nodules to O2 diffusion. J. Exp. Bot. 38, 1129–1140

    Google Scholar 

Download references

Author information

Author notes

  1. K. A. VandenBosch

    Present address: Biology Department, Texas A & M University, 77843-3258, College Station, TX, USA

Authors and Affiliations

  1. Department of Botany, University of Wisconsin, 53706, Madison, WI, USA

    K. A. VandenBosch & E. H. Newcomb

Authors
  1. K. A. VandenBosch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. H. Newcomb
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

VandenBosch, K.A., Newcomb, E.H. The occurrence of leghemoglobin protein in the uninfected interstitial cells of soybean root nodules. Planta 175, 442–451 (1988). https://doi.org/10.1007/BF00393063

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00393063

Key words

Search

Navigation