Skip to main content
Log in

Immunoelectron microscopy investigation of the cell surface of Azospirillum brasilense strains

  • Experimental Articles
  • Published:
Microbiology Aims and scope Submit manuscript

Abstract

The genus-specific surface protein antigens of Azospirillum brasilense strains were visualized immunochemically. The procedure used for cell sample preparation was optimized to ensure that the surface protein structures were detected on cells in situ. Gold and gold-silver nanoparticles were conjugated to antibodies raised against the flagellin of A. brasilense type strain Sp7, against the lipopolysaccharide of A. brasilense Sp245, and against the genus-specific protein determinants of A. brasilense Sp7. Electron microscopic analysis using nanoparticle-labeled antibodies revealed antigenic determinants of the polar flagellum on the A. brasilense Sp245 cell surface, which in these bacteria are normally screened from the surroundings by a lipopolysaccharide sheath. Pili-like structures were detected on the Sp245 wild-type strain and on its Fla Swa Omegon-Km mutant SK048, which are presumably involved in microcolonial spreading in these bacteria.

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

Access this article

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. and de-Bashan, L.E. How the plant growthpromoting bacterium Azospirillum promotes plant growth–a critical assessment, Adv. Agron., 2010, vol. 108, pp. 77–136.

    Article  CAS  Google Scholar 

  • Bashan, Y. and Levanony, H. Active attachment of Azospirillum brasilense Cd to quartz sand and to a light-textured soil by protein bridging, J. Gen. Microbiol., 1988, vol. 134, pp. 2269–2279.

    CAS  Google Scholar 

  • Burygin, G.L., Shirokov, A.A., Shelud’ko, A.V., Katsy, E.I., Shchygolev, S.Yu., and Matora, L.Yu., Detection of a sheath on Azospirillum brasilense polar flagellum, Microbiology (Moscow), 2007, vol. 76, pp. 728–734.

    Article  CAS  Google Scholar 

  • Collection of Rhizosphere Microorganisms of the Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences (IBPPM RAS) [Electronic document] (http://collection.ibppm.ru/). Verified 25.11.2016.

  • Croes, C.L., Moens, S., van Bastelaere, E., Vanderleyden, J., and Michiels, K.W., The polar flagellum mediated Azospirillum brasilense adsorption to wheat roots, J. Gen. Microbiol., 1993, vol. 139, pp. 2261–2269.

    Article  CAS  Google Scholar 

  • De Troch, P. and Vanderleyden, J., Surface properties and motility of Rhizobium and Azospirillum in relation to plant root attachment, Microb. Ecol., 1996, vol. 32, pp. 149–169.

    Article  PubMed  Google Scholar 

  • De Weger, L.A., Van der Vlug, C.I., Wijfjes, A.H.M., Bakker, P.A., Schippers, B., and Lugtenberg, B., Flagella of a plant-growth-stimulating Pseudomonas fluorescens strain are required for colonization of potato roots, J. Bacteriol., 1987, vol. 169, pp. 2769–2773.

    Article  PubMed  PubMed Central  Google Scholar 

  • Del Gallo, M., Negi, M., and Neira, C.A., Calcofluor- and lectin-binding exocellular polysaccharides of Azospirillum brasilense and Azospirillum lipoferum, J. Bacteriol., 1989, vol. 171, pp. 3504–3510.

    Article  PubMed  PubMed Central  Google Scholar 

  • Döbereiner, J. and Day, J.M., Associative symbioses in tropical grasses: characterization of microorganisms and dinitrogen-fixing sites, Proc. 1st Int. Symp. Nitrogen Fixation, Newton, W.E. and Nijmans, C.J., Eds., Pullman: Washington State Univ. Press, 1976, pp. 518–538.

    Google Scholar 

  • Dörr, J., Hurek, T., and Reinhold-Hurek, B., Type IV pili are involved in plant-microbe and fungus-microbe interactions, Mol. Microbiol., 1998, vol. 30, pp. 7–17.

    Article  PubMed  Google Scholar 

  • Frens, G., Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions, Nature Phys. Sci., 1973, vol. 241, no. 105, pp. 20–22.

    Article  CAS  Google Scholar 

  • Khanadeev, V.A., Khlebtsov, B.N., Staroverov, S.A., Vidyasheva, I.V., Skaptsov, A.A., Ileneva, E.S., Bogatyrev, V.A., Dykman, L.A., and Khlebtsov, N.G., Quantitative cell bioimaging using gold-nanoshell conjugates and phage antibodies, J. Biophotonics, 2011, vol. 4, nos. 1–2, pp. 74–83.

    Article  CAS  PubMed  Google Scholar 

  • Khlebtsov, B., Panfilova, E., Khanadeev, V., Bibikova, O., Terentyuk, G., Ivanov, A., Rumyantseva, V., Shilov, I., Ryabova, A., Loshchenov, V., and Khlebtsov, N., Nanocomposites containing silica-coated gold-silver nanocages and Yb-2, 4-dimethoxyhematoporphyrin: multifunctional capability of IR-luminescence detection, photosensitization, and photothermolysis, ACS Nano, 2011, vol. 5, pp. 7077–7089.

    Article  CAS  PubMed  Google Scholar 

  • Madi, L. and Henis, Y., Aggregation in Azospirillum brasilense Cd: conditions and factors involved in cell-to-cell adhesion, Plant Soil, 1989, vol. 115, pp. 89–98.

    Article  Google Scholar 

  • Madi, L., Kessel, M., Sadovnik, E., and Henis, Y., Electron microscopic studies of aggregation and pellicle formation in Azospirillum spp., Plant Soil, 1988, vol. 109, pp. 115–121.

    Article  Google Scholar 

  • Matora, L.Yu., Burygin, G.L., and Shchegolev, S.Yu., Study of immunochemical heterogeneity of Azospirillum brasilense lipopolysaccharides, Microbiology (Moscow), 2008, vol. 77, pp. 166–170.

    Article  CAS  Google Scholar 

  • Matora, L.Yu., Shvartsburd, B.I., and Shchegolev, S.Yu., Immunochemical analysis of O-specific polysaccharides from the soil nitrogen-fixing bacterium Azospirillum brasilense, Microbiology (Moscow), 1998, vol. 67, pp. 677–681.

    CAS  Google Scholar 

  • Michiels, K., Croes, C.L., and Vanderleyden, J., Two different modes of attachment of Azospirillum brasilense Sp7 to wheat roots, J. Gen. Microbiol., 1991, vol. 137, pp. 2241–2246.

    Article  CAS  Google Scholar 

  • Scheludko, A.V., Katsy, E.I., Ostudin, N.A., Gringaus, O.K., and Panasenko, V.I., Novel classes of Azospirillum brasilense mutants with defects in the assembly and functioning of polar and lateral flagella, Mol. Genet. Mikrobiol. Virusol., 1998, no. 4, pp. 33–37.

    Google Scholar 

  • Shelud’ko, A.V. and Katsy, E.I., Formation of polar bundles of pili and the behavior of Azospirillum brasilense cells in a semiliquid agar, Microbiology (Moscow), 2001, vol. 70, pp. 570–575.

    Article  Google Scholar 

  • Shelud’ko, A.V., Shirokov, A.A., Sokolova, M.K., Sokolov, O.I., Petrova, L.P., Matora, L.Yu., and Katsy, E.I., Wheat root colonization by Azospirillum brasilense strains with different motility, Microbiology (Moscow), 2010, vol. 79, pp. 688–695.

    Article  Google Scholar 

  • Shirokov, A.A., Krasov, A.I., Selivanov, N.Yu., Burygin, G.L., Shchyogolev, S.Yu., and Matora, L.Yu., Immunochemical detection of Azospirilla in soil with genus-specific antibodies, Microbiology (Moscow), 2015, vol. 84, pp. 263–267.

    Article  CAS  Google Scholar 

  • Skrabalak, S.E., Au, L., Li, X., and Xia, Y., Facile synthesis of Ag nanocubes and Au nanocages, Nat. Protoc., 2007, vol. 2, no. 9, pp. 2182–2190.

    Article  CAS  PubMed  Google Scholar 

  • Zhulin, I.B. and Armitage, J.P., The role of taxis in the ecology of Azospirillum, Symbiosis, 1992, vol. 13, pp. 199–206.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to L. Yu. Matora.

Additional information

Original Russian Text © A.A. Shirokov, A.A. Budanova, A.M. Burov, B.N. Khlebtsov, A.I. Krasov, S.Yu. Shchyogolev, L.Yu. Matora, 2017, published in Mikrobiologiya, 2017, Vol. 86, No. 4, pp. 476–482.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Shirokov, A.A., Budanova, A.A., Burov, A.M. et al. Immunoelectron microscopy investigation of the cell surface of Azospirillum brasilense strains. Microbiology 86, 487–492 (2017). https://doi.org/10.1134/S0026261717040142

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026261717040142

Keywords

Navigation