Skip to main content
SpringerLink
Log in

Enhanced nodule initiation on alfalfa by wild-typeRhizobium meliloti co-inoculated withnod gene mutants and other bacteria

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Nodule formation on alfalfa (Medicago sativa L.) roots was determined at different inoculum dosages for wild-typeRhizobium meliloti strain RCR2011 and for various mutant derivatives with altered nodulation behavior. The number of nodules formed on the whole length of the primary roots was essentially constant regardless of initial inoculum dosage or subsequent bacterial multiplication, indicative of homeostatic regulation of total nodule number. In contrast, the number of nodules formed in just the initially susceptible region of these roots was sigmoidally dependent on the number of wild-type bacteria added, increasing rapidly at dosages above 5·103 bacteria/plant. This behavior indicates the possible existence of a threshold barrier to nodule initiation in the host which the bacteria must overcome. When low dosages of the parent (103 cells/plant) were co-inoculated with 106 cells/plant of mutants lacking functionalnodA, nodC, nodE, nodF ornodH genes, nodule initiation was increased 10- to 30-fold. Analysis of nodule occupancy indicated that these mutants were able to help the parent (wild-type) strain initiate nodules without themselves occupying the nodules. Co-inoculation withR. trifolii orAgrobacterium tumefaciens cured of its Ti plasmid also markedly stimulated nodule initiation by theR. meliloti parent strain. Introduction of a segment of the symbiotic megaplasmid fromR. meliloti intoA. tumefaciens abolished this stimulation.Bradyrhizobium japonicum and a chromosomal Tn5 nod- mutant ofR. meliloti did not significantly stimulate nodule initiation when co-inoculated with wild-typeR. meliloti. These results indicate that certainnod gene mutants and members of theRhizobiaceae may produce extracellular “signals” that supplement the ability of wild-typeR. meliloti cells to induce crucial responses in the host.

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

EH:

emergent root hairs

kb:

kilobase

RDU:

relative distance unit

RT:

root tip

References

  • Bauer, W.D. (1981) Infection of legumes by rhizobia. Annu. Rev. Plant Physiol.32, 407–449

    Google Scholar 

  • Broughton, W.J. (1978) Control of specificity in legume-Rhizobium associations. J. Appl. Bacteriol.45, 165–194

    Google Scholar 

  • Banfalvi, Z., Sakanyan, V., Konzc, C., Kiss, A., Dusha, I., Kondorosi, A. (1981) Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid ofRhizobium meliloti. Mol. Gen. Genet.184, 318–325

    Google Scholar 

  • Bhagwat, A.A., Thomas, J. (1982) Legume-Rhizobium interactions: cowpea root exudate elicits faster nodulation response byRhizobium species. Appl. Environ. Microbiol.43, 800–805

    Google Scholar 

  • Bhuvaneswari, T.V., Bhagwat, A.A., Bauer, W.D. (1981) Transient susceptibility of root cells in four common legumes to nodulation by rhizobia. Plant Physiol.68, 1114–1149

    Google Scholar 

  • Bhuvaneswari, T.V., Lesniak, A.P., Bauer, W.D. (1988) Efficiency of nodule initiation in cowpea and soybean. Plant Physiol., in press

  • Bhuvaneswari, T.V., Mills, K.K., Crist, D.K., Evans, W.R., Bauer, W.D. (1983) Effects of culture age on symbiotic infectivity ofRhizobium japonicum. J. Bacteriol.153, 443–451

    Google Scholar 

  • Bhuvaneswari, T.V., Turgeon, G., Bauer, W.D. (1980) Early stages in the infection of soybean (Glycine max L. Merr.) byRhizobium japonicum. I. Localization of infectible root cells. Plant Physiol.66, 1027–1031

    Google Scholar 

  • Buikema, W.J., Long, S.R., Brown, S.E., van den Bos, R.C., Earl, C., Ausubel, F.M. (1983) Physical and genetic characterization ofRhizobium meliloti symbiotic mutants. J. Mol. Appl. Gen.2, 249–260

    Google Scholar 

  • Caetano-Anollés, G., Favelukes, G. (1986) Host-symbiont specificity expressed during early adsorption ofRhizobium meliloti to the root surface of alfalfa. Appl. Environ. Microbiol.52, 377–382

    Google Scholar 

  • Caetano-Anoll'es, G., Wall, L.G., DeMicheli, A.T., Macchi, E.M., Favelukes, G. (1985) Attachment of nonmotile and nonchemotactic mutants ofRhizobium meliloti to alfalfa roots. In: Nitrogen fixation research progress, p. 257, Evans, H.J., Bottomley, P.J., Newton, W.E., eds. Martinus Nijhoff, Boston

    Google Scholar 

  • Caetano-Anollés, G., Wall, L.G., DeMicheli, A.T., Macchi, E.M., Bauer, W.D., Favelukes, G. (1988) Role of motility and chemotaxis in efficiency of nodulation byR. meliloti. Plant Physiol., in press

  • Calvert, H.E., Pence, M.K., Pierce, M., Malik, N.S.A., Bauer, W.D. (1984) Anatomical analysis of the development and distribution ofRhizobium infections in soybean roots. Can. J. Bot.62, 2375–2384

    Google Scholar 

  • Debelle, F., Rosenberg, C., Vasse, J., Maillet, F., Martinez, E., Denarie, J., Truchet, G. (1986) Assignment of symbiotic developmental phenotypes to common and specific nodulation (nod) genetic loci ofRhizobium meliloti. J. Bacteriol.168, 1075–1086

    Google Scholar 

  • Debelle, F., Sharma, S.B. (1986) Nucleotide sequence ofRhizobium meliloti RCR2011 genes involved in host specificity of nodulation. Nucleic Acids Res.14, 7453–7472

    Google Scholar 

  • Djordjevic, S.P., Chen, H., Batley, M., Redmond, J.W., Rolfe, B.G. (1987) Nitrogen fixation ability of exopolysaccharide synthesis mutants ofRhizobium sp. strain NGR234 andRhizobium trifolii is restored by the addition of homologous exopolysaccharides. J. Bacteriol.169, 53–60

    Google Scholar 

  • Egelhoff, T.T., Fisher, R.F., Jacobs, T.W., Mulligan, J.T., Long, S.R. (1985) Nucleotide sequence ofRhizobium meliloti 1021 nodulation genes:nodD is read divergently fromnod ABC. DNA4, 241–248

    Google Scholar 

  • Favelukes, G., Caetano-Anollés, G. (1985)Rhizobium meliloti specifically attached to alfalfa roots: obligatory precursor towards nodulation. In: Nitrogen fixation research progress, p. 259, Evans, H.J., Bottomley, P.J., Newton, W.E., eds., Martinus Nijhoff, Boston

    Google Scholar 

  • Fischer, R.F., Tu, J.K., Long, S.R. (1985) Conserved nodulation genes inRhizobium meliloti andRhizobium trifolii. Appl. Environ. Microbiol.49, 1432–1435

    Google Scholar 

  • Halverson, L.J., Stacey, G. (1984) Host recognition in theRhizobium-soybean symbiosis. Detection of a protein factor in soybean root exudate which is involved in the nodulation process. Plant Physiol.74, 84–89

    Google Scholar 

  • Halverson, L.J., Stacey, G. (1985) Host recognition in theRhizobium-soybean symbiosis. Evidence for the involvement of lectin in nodulation. Plant Physiol.77, 621–625

    Google Scholar 

  • Halverson, L.J., Stacey, G. (1986a) Effect of lectin on nodulation by wild-typeBradyrhizobium japonicum and a nodulation-defective mutant. Appl. Environ. Microbiol.51, 753–760

    Google Scholar 

  • Halverson, L.J., Stacey, G. (1986b) Signal exchange in plantmicrobe interactions. Microbiol. Rev.50, 193–225

    Google Scholar 

  • Heron, D.S., Pueppke, S.G. (1987) Regulation of nodulation in the soybean-Rhizobium symbiosis. Strain and cultivar variability. Plant Physiol.84, 1391–1396

    Google Scholar 

  • Hirsch, A.M., Long, S.R., Bang, M., Haskins, N., Ausubel, F.M. (1982) Structural studies of alfalfa roots infected with nodulation mutants ofRhizobium meliloti. J. Bacteriol.151, 411–419

    Google Scholar 

  • Hoagland, D.R., Arnon, D.I. (1950) The water-culture method for growing plants without soil. (Calif. Agric. Exp. Sta. Circ. No. 347 (rev. edn.))

  • Horvath, B., Kondorosi, E., John, M., Schmidt, J., Torok, I., Gyorgypal, Z., Barabas, I., Wieneke, U., Schell, J., Kondorosi, A. (1986) Organization, structure and symbiotic functions ofRhizobium meliloti nodulation genes determining host specificity for alfalfa. Cell46, 335–343

    Google Scholar 

  • Jacobs, T.W., Egelhoff, T.T., Long, S.R. (1985) Physical and genetic map ofRhizobium meliloti nodulation gene region and nucleotide sequence ofnodC. J. Bacteriol.162, 469–476

    Google Scholar 

  • Jensen, H.L. (1942) Nitrogen fixation in leguminous plants. I. General characters of root nodule bacteria isolated from species ofMedicago andTrifolium in Australia. Proc. Linn. Soc. N.S.W.66, 98–108

    Google Scholar 

  • Kondorosi, E., Banfalvi, Z., Kondorosi, A. (1984) Physical and genetic analysis of a symbiotic region ofRhizobium meliloti: identification of nodulation genes. Mol. Gen. Genet.193, 445–452

    Google Scholar 

  • Long, S.R., Buikema, W.J., Ausubel, F.M. (1982) Cloning ofRhizobium meliloti nodulation genes by direct complementation of Nod- mutants. Nature298, 318–325

    Google Scholar 

  • Meade, H.M., Long, S.R., Ruvkun, G.B., Brown, S.E., Ausubel, F.M. (1982) Physical and genetic characterization of symbiotic and auxotrophic mutants ofRhizobium meliloti induced by transposon mutagenesis. J. Bacteriol.149, 114–122

    Google Scholar 

  • Ott, L. (1984) An introduction to statistical methods and data analysis, 2nd edn. Duxbury Press, Boston, Mass., USA

    Google Scholar 

  • Pierce, M., Bauer, W.D. (1983) A rapid regulatory response governing nodulation in soybean. Plant Physiol.73, 286–290

    Google Scholar 

  • Putnoky, P., Kondorosi, A. (1986) Two gene clusters ofRhizobium meliloti code for essential nodulation functions and a third influences nodulation efficiency. J. Bacteriol.167, 881–887

    Google Scholar 

  • Renalier, M.H., Batut, J., Ghai, J., Terzaghi, B., Gherardi, M., David, M., Garnerone, A.M., Vasse, J., Truchet, G., Huguet, T., Boistard, P. (1987) A new symbiotic cluster on the pSym megaplasmid ofRhizobium meliloti 2011 carries a functionalfix gene repeat and anod locus. J. Bacteriol.169, 2231–2238

    Google Scholar 

  • Rodriguez-Quinones, F., Banfalvi, Z., Murphy, P., Kondorosi, A. (1987) Interspecies homolog of nodulation genes inRhizobium. Plant Mol. Biol.8, 61–75

    Google Scholar 

  • Rolfe, B.G., Gresshoff, P.M., Shine, J., Vincent, J.M. (1980) Interaction between a non-nodulating and an ineffective mutant ofRhizobium trifolii resulting in effective (nitrogenfixing) nodulation. Appl. Environ. Microbiol.39, 449–452

    Google Scholar 

  • Rosenberg, C., Boistard, P., Dénarié, J., Casse-Delbart, F. (1981) Genes controlling early and late functions in symbiosis are located on a megaplasmid inRhizobium meliloti. Mol. Gen. Genet.184, 326–333

    Google Scholar 

  • Sargent, L., Huang, S.Z., Rolfe, B.G., Djordjevic, M.A. (1987) Split-root assays usingTrifolium subterraneum show thatRhizobium infection induces a systemic response that can inhibit nodulation of another invasiveRhizobium strain. Appl. Environ. Microbiol.53, 1611–1619

    Google Scholar 

  • Takats, S.T. (1986) Suppression of nodulation in soybeans by superoptimal inoculation withBradyrhizobium japonicum. Physiol. Plant.66, 669–673

    Google Scholar 

  • Truchet, G., Debellé, F., Vasse, J., Terzaghi, B., Garnerone, A.M., Rosenberg, C., Batut, J., Maillet, F., Dénarié, J. (1985) Identification of aRhizobium meliloti pSym 2011 region controlling the host specificity of root hair curling and nodulation. J. Bacteriol.164, 1200–1210

    Google Scholar 

  • Truchet, G., Rosenberg, C., Vasse, J., Julliot, J.S., Camut, S., Dénarié, J. (1984) Transfer ofRhizobium meliloti pSym genes intoAgrobacterium tumefaciens. Host-specific nodulation by atypical infection. J. Bacteriol.157, 134–142

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Agronomy, Ohio State University, 2021 Coffey Road, 43210-1086, Columbus, OH, USA

    Gustavo Caetano-Anollés & Wolfgang D. Bauer

Authors
  1. Gustavo Caetano-Anollés
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfgang D. Bauer
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This is journal article No. 188-87 of the Ohio Agricultural Research and Development Center

Rights and permissions

Reprints and permissions

About this article

Cite this article

Caetano-Anollés, G., Bauer, W.D. Enhanced nodule initiation on alfalfa by wild-typeRhizobium meliloti co-inoculated withnod gene mutants and other bacteria. Planta 174, 385–395 (1988). https://doi.org/10.1007/BF00959525

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00959525

Key words

Search

Navigation