Skip to main content
SpringerLink
Log in

Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules

  • Original Paper
  • Published:
Biology and Fertility of Soils Aims and scope Submit manuscript

Abstract

Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH +4 -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N2 fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O2 than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH +4 -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.

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

References

  • Bashan Y, Levanony H (1988) Interaction between Azospirillum brasilense CD and wheat root cells during early stages of root colonization. In: Klingmüller W (ed) Azospirillum IV. Springer Verlag, Berlin, pp 166–173

    Google Scholar 

  • Bergersen FJ (1982) Root nodules of legumes: Structure and functions. Research Studies Press, Wiley, Chichester

    Google Scholar 

  • Bender GL, Preston L, Bernard D, Rolfe BG (1988) Formation of nodule-like structures on the roots of the non-legumes rice and wheat. In: Gresskoff PM, Roith G, Stacey G, Newton WE (eds) Nitrogen fixation: Achievements and objectives. Chapman and Hall, New York, p 825

    Google Scholar 

  • Boddey RM, Urquiaga S, Reis V, Döbereiner J (1991) Biological nitrogen fixation associated with sugar cane. Plant and Soil 137:111–117

    Google Scholar 

  • Christiansen-Weniger C (1992a) N2 fixation by ammonium-excreting Azospirillum brasilense in auxine-induced root tumours of wheat (Triticum aestivum L.). Biol Fertil Soils 13:165–172

    Google Scholar 

  • Christiansen-Weniger C (1992b) Dynamics of a transposon Tn 5 mutant of Azospirillum brasilense in soil and rhizosphere of spring wheat. Symbiosis 13:85–100

    Google Scholar 

  • Christiansen-Weniger C, Van Veen JA (1991a) Nitrogen fixation by Azospirillum brasilense in soil and the rhizosphere under controlled environmental conditions. Biol Fertil Soils 12:100–106

    Google Scholar 

  • Christiansen-Weniger C, Van Veen JA (1991b) NH +4 -excreting Azospirillum brasilense mutants enhance the nitrogen supply of a wheat host. Appl Environ Microbiol 57:3006–3012

    Google Scholar 

  • Christiansen-Weniger C, Boddey RM, Döbereiner J (1985) Evaluation of nitrogen fixation in sorghum cultivars inoculated with different strains of Azospirillum spp. In: Klingmüller W (ed) Azospirillum III. Springer Verlag, Berlin, pp 180–188

    Google Scholar 

  • Christiansen-Weniger C, Groneman AF, Van Veen JA (1992) Associative N2-fixation and root exudation of organic acid from wheat cultivars of different aluminium tolerance. Plant and Soil 139:167–174

    Google Scholar 

  • Döbereiner J, Day JM (1976) Associative symbiosis in tropical grasses. Characterisation of microorganisms and dinitrogen fixation sites. In: Newton WE, Nyman CJ (eds) Proc 1st Int Symp on Nitrogen Fixation, Vol 2, Washington State University Press, Washington, p 518

    Google Scholar 

  • Echlin P (1991) Low temperature microscopy and analysis of plant material. In: Boekestein A (ed) Past, present and future of electron microscopy in agricultural research, Dienst Landbouwkundig Onderzoek DLO, The Netherlands, pp 75–85

    Google Scholar 

  • Fahreus G (1957) The infection of clover root hairs by nodule bacteria studied by a simple glass slide technique. J Gen Microbiol 16:374–381

    Google Scholar 

  • Gantar M, Kerby NW, Rowell P, Obreht Z (1991) Colonization of wheat (Triticum vulgare L.) by N2 fixing cyanobacteria: I. A survey of soil cyanobacterial isolates forming associations with roots. New Phytol 118:477–483

    Google Scholar 

  • Hu CY, Chee PP, Chesney RH, Zhou JH, Miller PD, O'Brien WT (1990) Intrinsic GUS-like activities in seed plants. Plant Cell Rep 9:1–5

    Google Scholar 

  • James EK, Sprent JI, Sutherland JM, Mcinroy SG, Minchin FR (1992) The structure of nitrogen fixing root nodules on the aquatic mimosid legume Neptunia plena. Ann Bot 69:173–180

    Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    Google Scholar 

  • Jefferson RA, Burgess SM, Hirsh D (1986) Beta glucuronidase from Escherichia coli as gene fusion marker. Proc Natl Acad Sci USA 83:8447–8451

    Google Scholar 

  • Kleiner D (1984) Bakterien und Ammonium. Forum Mikrobiol 7:13–19

    Google Scholar 

  • Lalonde M, Devoe IW (1975) Scanning electron microscopy of the Alnus crispa var. mollis Fern. root nodule endophyte. Arch Microbiol 105:87–94

    Google Scholar 

  • Longeri L, Mireya Abarzua (1989) Ultrastructure of Francia isolated from three Chilean shrubs (Rhamnaceae). In: Skinner FA, Boddey RM, Fendrik I (eds) Nitrogen fixation with non-legumes, Kluwer Academic Publishers, Dordrecht Boston London, pp 47–53

    Google Scholar 

  • Machado HB, Fumayama S, Rigo LV, Pedrosa FO (1991) Excretion of ammonium by Azospirillum brasilense mutants resistant to ethylene-diamine. Can J Microbiol 37:549–553

    Google Scholar 

  • Malik KA, Bilal R (1988) Survival and colonization of inoculated bacteria in kallar grass rhizosphere and quantification of N2-fixation. Plant and Soil 110:329–338

    Google Scholar 

  • Michiels KW, Verreth C, Vanderleyden J (1990) Azospirillum lipoferum and Azospirillum brasilense surface polysaccharides mutants that are affected in flocculation. J Appl Bacteriol 69:705–711

    Google Scholar 

  • Michiels KW, Croes CL, Vanderleyden J (1991) Two different modes of attachment of Azospirillum brasilense Sp 7 to wheat roots. J Gen Microbiol 137:2241–2246

    Google Scholar 

  • Nap JP, Bisseling T (1990) Developmental biology of a plant-procaryote symbiosis: The legume root nodule. Science 250:948–954

    Google Scholar 

  • Nap JP, Van de Wiel C, Spaink HP, Moerman M, Van den Heuvel M, Djordjevic MA, Van Lammeren AAM, Van Kammen A, Bisseling T (1989) The relationship between nodulin gene expression and the Rhizobium nod genes in Vicia sativa root nodule development. Mol Plant-Microbe Interactions 2:53

    Google Scholar 

  • Nayak PNM, Ladha JK, Watanabe I (1986) The fate of marker Azospirillum lipoferum inoculated into rice and its effect on growth, yield and N2-fixation studied by acetylene reduction, 15N2-feeding and 15N-dilution technique. Biol Fertil Soils 2:7–14

    Google Scholar 

  • Nur I, Okon Y, Henis Y (1980) An increase in nitrogen content of Setaria italica and Zea maize inoculated with Azospirillum. Can J Microbiol 26:482–486

    Google Scholar 

  • Okon Y, Heitler PG, Hardy R (1983) N2-fixation by Azospirillum brasilense and its incorporation into the host Setaria italica. Appl Environ Microbiol 46:695–697

    Google Scholar 

  • Patriquin DG, Döbereiner J (1978) Light microscopy observation of tetrazolium reducing bacteria in the endorhizosphere of maize and other grasses in brazil. Can J Microbiol 24:734–742

    Google Scholar 

  • Reding HK, Hartel PG, Wiegel J (1991) Effect of Xantobacter, isolated and characterized from rice roots, on growth of wetland rice. Plant and Soil 138:221–229

    Google Scholar 

  • Spaink HP, Sheeley DM, VanBrussel AAN, Glushka J, York WS, Tak T, Geiger O, Kennedy EP, Reinhold VN, Lugtenberg BJJ (1991) A novel highly unsaturated fatty acid moeity of lipooligosaccharide signals determines host specificity of Rhizobium. Nature (London) 354:125

    Google Scholar 

  • Tchan YT, Kennedy IR (1989) Possible N2-fixing root nodules induced in non-legumes. Agric Sci (AIAS, Melbourne) 2:57–59

    Google Scholar 

  • Tchan YT, Zeman AMM, Kennedy IR (1991) Nitrogen fixation in para-nodules of wheat roots by introduced free living diazotrophs. Plant and Soil 137:43–47

    Google Scholar 

  • Trinick MJ, Hadobas PA (1988) Biology of Parasponia-Bradyrhizobium symbiosis. Plant and Soil 110:177–185

    Google Scholar 

  • Truchet G, Roche P, Lerouge P, Vasse J, Camut S, de Billy F, Prome JC, Denarie J (1991) Sulphated lipo-oligosaccharide signals of Rhizobium meliloti elicit root nodule organogenesis in Alfalfa. Nature (London) 351:670–673

    Google Scholar 

  • Watanabe I, Lin C (1983) Response of wetland rice to inoculation with Azospirillum lipoferum and Pseudomonas sp. Soil Sci Plant Nutr 30:117–124

    Google Scholar 

  • Whallon JH, Acker G, El-Khawas H (1985) Electron microscopy of young wheat roots inoculated with Azospirillum. In: Klingmüller W (ed) Azospirillum III. Springer Verlag, Berlin, pp 221–229

    Google Scholar 

  • You CB, Song W, Wang HX, Li PJ, Lin M, Hai WL (1991) Association of Alcaligenes faecalis with wetland rice. Plant and Soil 137:81–85

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. F. A. Janssens Laboratory of Genetics, Catholic University of Leuven, Willem de Croylaan 42, B-3001, Heverlee, Belgium

    C. Christiansen-Weniger & J. Vanderleyden

Authors
  1. C. Christiansen-Weniger
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Vanderleyden
    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

Christiansen-Weniger, C., Vanderleyden, J. Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules. Biol Fert Soils 17, 1–8 (1994). https://doi.org/10.1007/BF00418663

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00418663

Key words

Search

Navigation