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The occurrence of unusual laminated structures rich in β-1,4-glucans in plastids of Phaseolus vulgaris root-nodule cells infected by an ineffective C4-dicarboxylic-acid mutant of Rhizobium leguminosarum bv. phaseoli

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Abstract

Root nodules induced in Phaseolus vulgaris L. by the wild-type (WT) and a C4-dicarboxylic-acid mutant strain of Rhizobium leguminosarum biovar. phaseoli were compared on the basis of ultrastructure and cytochemistry of cellulose subunits. The mutant bacteroids failed to colonize infected host cells in a normal manner, and presented a premature degenerative appearance. Starch granules, rough endoplasmic reticulum and mitochondria were found to accumulate in the ineffective nodules. The most striking difference between effective and ineffective nodules was the presence of unusual spherical, laminated structures in plastids of mutant-infected host cells only. Cytochemical observations showed that these structures contain β-1,4-glucans. The presence of β-1,4-glucans within such structures may be caused by the activity of a cellulase which is produced by either the bacteroids or the host cell and is locally hydrolyzing the host cell-wall, thus releasing cellulose subunits into the cytoplasm. Another possibility is denovo synthesis of β-1,4-glucans in the host cell.

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Abbreviations

PBS:

phosphate-buffered saline

PEG:

polyethylene glycol

PHB:

poly-β-hydroxybutyrate

RER:

rough endoplasmic reticulum

TTC:

2,3,5-triphenyl tetrazolium chloride

WT:

wild-type

B :

bacteroid

C :

cortex

Cy :

cytoplasm

CC :

central core

HCW :

host cell-wall

IC :

infected cell

IS :

inter-cellular space

LS :

laminated structure

N :

nucleus

P :

peroxisome

PHB :

poly-β-hydroxybutyrate

Pl :

plastid

PlM :

plastid matrix

PM :

peribacteroid membrane

RER :

rough endoplasmic reticulum

S :

starch grain

UC :

uninfected cell

Va :

vacuole

VB :

vascular bundle

References

  • Arwas, R., McKay, I.A., Rowney, F.R.P., Dilworth, M.J., Glenn, A.R. (1985) Properties of organic acid utilization mutants of Rhizobium leguminosarum strain 300. J. Gen. Microbiol. 131, 2059–2066

    Google Scholar 

  • Baird, L.M., Webster, B.D. (1982) Morphogenesis of effective and ineffective root nodules in Phaseolus vulgaris. L. Bot. Gaz. 143, 41–51

    Google Scholar 

  • Benhamou, N. (1989) Cytochemical localization of β-(1→ 4)-d-glucans in plant and fungal cells using an exoglucanase-gold complex. Electron Microsc. Rev. 2, 123–138

    Google Scholar 

  • Benhamou, N., Chamberland, H., Ouellette, G.B., Pauzé, F.J. (1987) Ultrastructural localization of β-(1→4)-d-glucans in two pathogenic fungi and in their host tissues by means of an exoglucanase-gold complex. Can. J. Microbiol. 33, 405–417

    Google Scholar 

  • Bergersen, F.J. (1957) The structure of ineffective nodules of legumes: an unusual type of ineffectiveness and an appraisal of present knowledge. Aust. J. Biol. Sci. 10, 233–242

    Google Scholar 

  • Berghem, L.E.R., Pettersson, L.G. (1973) The mechanism of enzymatic cellulose degradation. Purification of a cellulocytic enzyme from Trichoderma viride active on highly ordered cellulose. Eur. J. Biochem. 37, 21–30

    Google Scholar 

  • Berghem, L.E.R., Pettersson, L.G., Axiö-Fredricksson, U.-B. (1975) The mechanism of enzymatic cellulose degradation. Characterization and enzymatic properties of a β-1,4-glucan cellobiohydrolase from Trichoderma viride. Eur. J. Biochem. 53, 55–62

    Google Scholar 

  • Chalifour, F.-P., Benhamou, N. (1989) Indirect evidence for cellulase production by Rhizobium in pea root nodules during bacteroid differentiation: cytochemical aspects of cellulose breakdown in rhizobial droplets. Can. J. Microbiol. 35, 821–829

    Google Scholar 

  • Dazzo, F.B., Truchet, G.L. (1983) Interactions of lectins and their saccharide receptors in the Rhizobium-legume symbiosis. J. Membr. Biol. 73, 1–16

    Google Scholar 

  • Doherty, D., Leigh, J.A., Glazebrook, J., Walker, G.C. (1988) Rhizobium meliloti mutants that overproduce the R. meliloti acidic calcofluor-binding exopolysaccharide. J. Bacteriol. 170, 4249–4256

    Google Scholar 

  • Finan, T.M., Wood, J.M., Jordan, D.C.. (1983) Symbiotic properties of C4-dicarboxylic acid transport mutants of Rhizobium leguminosarum. J. Bacteriol. 154, 1403–1413

    Google Scholar 

  • Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold solutions. Nature (London), Phys. Sci. 241, 20–22

    Google Scholar 

  • Huber, J.D., Parker, F., Odland, G.F. (1968) A basic fuchsin and alkalinized methylene blue rapid stain for epoxy-embedded tissue. Stain Technol. 43, 83–87

    Google Scholar 

  • Lafontaine, P.J., Lafrenière, C., Antoun, H. (1989a) Some properties of carbohydrate and C4-dicarboxylic acid utilization negative mutants of Rhizobium leguminosarum biovar phaseoli strain P121. Plant Soil 120, 195–201

    Google Scholar 

  • Lafontaine, P.J., Lafrenière, C., Chalifour, F.-P., Dion, P., Antoun, H. (1989b) Carbohydrate and organic acid composition of effective and ineffective root nodules of Phaseolus vulgaris Physiol. Plant. 76, 507–513

    Google Scholar 

  • Lafrenière, C., Lafontaine, P.J., Marion, C., Antoun, H. (1987) Oxidation of substrate in organic acids utilization negative mutants and the wild type Rhizobium meliloti strain S14. Plant Soil 101, 73–78

    Google Scholar 

  • Lalande, R., Antoun, H., Paré, T., Joyal, P. (1986) Effets de l'inoculation avec des sourches du Rhizobium leguminosarum biovar phaseoli sur le rendement et la teneur en azote du haricot (Phaseolus vulgaris). Naturaliste Can. (Rev. Ecol. Syst.) 113, 337–346

    Google Scholar 

  • Leigh, J.A. Lee, C.C. (1988) Characterization of polysaccharides of Rhizobium meliloti exo mutants that form ineffective nodules. J. Bacteriol. 170, 3327–3332

    Google Scholar 

  • Leigh, J.A., Reed, J.W., Hanks, J.F., Hirsh, A.M., Walker, G.C. (1987) Rhizobium meliloti mutants that fail to succinylate their calcofluor-binding exopolysaccharide are defective in nodule invasion. Cell 51, 579–587

    Google Scholar 

  • Long, S.R. Cooper, J. (1988) Overview of symbiosis. In: Proc. 4th Int. Symp. Mol. Gen. Plant Microbe Interact., Acapulo, Mexico, pp. 163–177, Palacios, R., Verma, D.P.S., eds. American Phytopathological Society, Press, St. Paul, Minn., USA

    Google Scholar 

  • Long, S., Reed, J.W., Himawan, J., Walker, G.C. (1988) Genetic analysis of a cluster of genes required for synthesis of the calcofluor-binding exopolysaccharide of Rhizobium meliloti. J. Bacteriol. 170, 4239–4248

    Google Scholar 

  • Martinez-Molina, E., Morales, V.M., Hubbell, D.H. (1979) Hydrolytic enzyme production by Rhizobium. Appl. Environ. Microbiol. 38, 1186–1188

    Google Scholar 

  • MacKenzie, C.R., Jordan, D.C. (1974) Ultrastructure of root nodules formed by ineffective strains of Rhizobium meliloti. Can. J. Microbiol. 20, 755–758

    Google Scholar 

  • Newcomb, W., Syono, K., Torrey, J.G. (1977) Development of an ineffective pea root nodule: morphogenesis, fine structure, and cytokinin biosynthesis. Can. J. Bot. 55, 1891–1907

    Google Scholar 

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

    Google Scholar 

  • Nogueira, N.D.L., Da Silva, D.M., Saito, S.M.T. (1979) Estudo ao microscopio eletrônico de nodulos em Phaseolus vulgaris L. causados por uma estripe inefetiva de Rhizobium. Turrialba 29, 93–96

    Google Scholar 

  • Pankhurst, C.E. (1974) Ineffective Rhizobium trifolii mutants examined by immune-diffusion, gel-electrophoresis and electron microscopy. J. Gen. Microbiol. 82, 405–413

    Google Scholar 

  • Ronson, C.W., Lyttleton, P., Robertson, J.G. (1981) C4-dicarboxylate transport mutants of Rhizobium trifolii form ineffective nodules on Trifolium repens. Proc. Natl. Acad. Sci. USA 78, 4284–4288

    Google Scholar 

  • Selvaraj, G., Hooper, I., Shantharam, S., Iyer, V.N., Barran, L., Wheatcroft, R., Watson, R.J. (1987) Derivation and molecular characterization of symbiotically deficient mutants of Rhizobium meliloti. Can. J. Microbiol. 33, 739–747

    Google Scholar 

  • Van den Bosch, K.A., Noël, K.D., Kaneko, Y., Newcomb, E.H. (1985) Nodule initiation elicited by noninfective mutants of Rhizobium phaseoli. J. Bacteriol. 162, 950–959

    Google Scholar 

  • Watson, R.J., Chan, Y.-K., Wheatcroft, R., Yang, A.-F., Han, S. (1988) Rhizobium meliloti genes required for C4-dicarboxylate transport and symbiotic nitrogen fixation are located on a megaplasmid. J. Bacteriol. 170, 927–934

    Google Scholar 

  • Webb, M.A., Newcomb, E.H. (1987) Cellular compartimentation of ureide biogenesis in root nodules of cowpea (Vigna unguiculata (L.) Walp.). Planta 172, 162–175

    Google Scholar 

  • Werner, D., Mörschel, E. (1978) Differentiation of nodules of Glycine max. Ultrastructural studies of plant cells and bacteroids. Planta 141, 169–177

    Google Scholar 

  • Werner, D., Mörschel, E., Kort, R., Mellor, R.B., Bassarab, S. (1984) Lysis of bacteroids in the vicinity of the host cell nucleus in an ineffective (fix−) root nodule of soybean (Glycine max). Planta 162, 8–16

    Google Scholar 

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Author notes

  1. Pierre J. Lafontaine

    Present address: Institut botanique de l'Université de Montréal, 4101 rue Sherbrooke Est, H1X 2B2, Montréal, Qué., Canada

Authors and Affiliations

  1. Département des Sols, Université Lavai, G1K 7P4, Québec, Canada

    Pierre J. Lafontaine & Hani Antoun

  2. Département de Phytologie, Université Lavai, G1K 7P4, Québec, Canada

    Nicole Benhamou

Authors
  1. Pierre J. Lafontaine
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  2. Nicole Benhamou
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  3. Hani Antoun
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Additional information

This work was supported by grants from the Natural Sciences and Engineering Research Council of Canada to H. Antoun and N. Benhamou. The authors are grateful to Dr. E.H. Newcomb, Department of Botany, University of Wisconsin, Madison, USA, for helpful comments on the interpretation of the micrographs. We also wish to thank S. Noël and H. Bissonnette for excellent technical assistance, Mrs. M. Pelchat for typing this manuscript, and Dr. G.B. Ouellette, Centre Forestier des Laurentides, Agriculture Canada, Sainte-Foy, Que., Canada, for the use of his light microscope. Printing for this paper was supported by the Conseil des recherches et services agricoles du Québec.

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Lafontaine, P.J., Benhamou, N. & Antoun, H. The occurrence of unusual laminated structures rich in β-1,4-glucans in plastids of Phaseolus vulgaris root-nodule cells infected by an ineffective C4-dicarboxylic-acid mutant of Rhizobium leguminosarum bv. phaseoli . Planta 180, 312–323 (1990). https://doi.org/10.1007/BF00198782

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  • DOI: https://doi.org/10.1007/BF00198782

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