World Journal of Microbiology and Biotechnology

, Volume 21, Issue 8–9, pp 1591–1597 | Cite as

Antifungal and Root Surface Colonization Properties of GFP-Tagged Paenibacillus brasilensis PB177

  • I. von der Weid
  • V. Artursson
  • L. Seldin
  • J. K. Jansson


This study evaluates the potential of Paenibacillus brasilensis strain PB177 to inhibit phytopathogenic fungi commonly causing maize diseases and to colonize maize plants. In vitro assays demonstrated antagonistic activity against the fungal pathogens, Fusarium moniliforme and Diplodia macrospora. The PB177 strain was tagged with the gfp gene, encoding the green fluorescent protein (GFP) and GFP-tagged bacteria were detected attached to maize roots by stereo- and confocal microscopy. The GFP-tagged bacteria were also used to treat maize seeds before challenging the seeds with two phytopathogenic fungi. The results demonstrated that the bacterial cells are mobilized to the maize roots in the presence of the fungal pathogens. The ability of P. brasilensis PB177 to inhibit fungal growth in vitro and its capability of colonization of maize roots in vivo suggest a potential application of this strain as a biological control agent. This is the first report on the successful introduction of the GFP marker gene into a P. brasilensis strain, enabling the direct observation of these promising plant growth promoting bacteria on maize roots in situ.


Antagonism antifungal substances biological control GFP maize colonization Paenibacillus brasilensis 


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  1. Ash, C., Priest, F.G., Collins, M.D. 1993Molecular identification of rRNA group 3 bacilli (Ash, Farrow, Wallbanks & Collins) using a PCR probe testAntonie van Leeuwenhoek64253260Google Scholar
  2. Assmus, B., Hutzler, P., Kirchhof, G., Amann, R., Lawrence, J.K., Hartmann, A. 1995In situ localization of Azospirillum brasilense in the rhizosphere of wheat with fluorescently labelled rRNA-target oligonucleotide probes and scanning confocal laser microscopyApplied and Environmental Microbiology6110131019Google Scholar
  3. Axelrood, P.E., Chow, M.L., Arnold, C.S., Lu, K., McDermott, J.M., Davies, J. 2002Cultivation-dependent characterization of bacterial diversity from British Columbia forest soils subjected to disturbanceCanadian Journal of Microbiology4864354Google Scholar
  4. Beatty, P.H., Jensen, S.E. 2002Paenibacillus polymyxa produces fusaricidin-type antifungal antibiotics active against Leptosphaeria maculans, the causative agent of blackleg disease of canolaCanadian Journal of Microbiology48159169CrossRefGoogle Scholar
  5. Bent, E., Chanway, C.P. 2002Potential for misidentification of a spore-forming Paenibacillus polymyxa isolated as an endophyte by using culture-based methodsApplied and Environmental Microbiology6846504652CrossRefGoogle Scholar
  6. Berge, O., Guinebretiere, M.H., Achouak, W., Normand, P., Heulin, T. 2002Paenibacillus graminis sp. nov. and Paenibacillus odorifer sp. nov., isolated from plant roots, soil and foodInternational Journal of Systematic and Evolutionary Microbiology52607616Google Scholar
  7. Budi, S.W., Tuinen, D., Arnould, C., Dumas-Gaudot, E., Gianinazzi-Pearson, V., Gianinazzi, S. 2000Hydrolytic enzyme activity of Paenibacillus sp. strain B2 and effects of the antagonistic bacterium on cell integrity of two soil-born pathogenic fungiApplied Soil Ecology15191199CrossRefGoogle Scholar
  8. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W., Prasher, D.C. 1994Green fluorescent protein as a marker for gene expressionScience263802805Google Scholar
  9. Chelius, M.K., Triplett, E.W. 2000Immunolocalization of dinitrogenase reductase produced by Klebsiela pneumoniae in association with Zea mays LApplied and Environmental Microbiology66783787CrossRefGoogle Scholar
  10. Cormack, B.P., Valdivia, R.H., Falkow, S. 1996FACS-optimized mutants of the green fluorescent protein (GFP)Gene1733338CrossRefGoogle Scholar
  11. Dandurand, L.M., Schotzko, D.J., Knudsen, G.R. 1997Spatial patterns of rhizoplane populations of Pseudomonas fluorescensApplied and Environmental Microbiology6332113217Google Scholar
  12. Dijksterhuis, J., Sanders, M., Gorris, L.G., Smid, E.J. 1999Antibiosis plays a role in the context of direct interaction during antagonism of Paenibacillus polymyxa towards Fusarium oxysporumJournal of Applied Microbiology861321CrossRefGoogle Scholar
  13. Fortineau, N., Trieu-cuot, P., Gaillot, O., Pellegrini, E., Berche, P., Gaillard, J.-L. 2000Optimization of green fluorescent protein expression vectors for in vitro and in vivo detection of Listeria monocytogenesResearch in Microbiology151353360CrossRefGoogle Scholar
  14. Garbeva, P., Overbeek, L.S., Vuurde, J.W.L., Elsas, J.D. 2001Analysis of endophytic bacterial communities of potato by planting and denaturing gradient gel electrophoresis (DGGE) of 16S rDNA based PCR fragmentsMicrobial Ecology41369383Google Scholar
  15. Garbeva, P., Veen, J.A., Elsas, J.D. 2003Predominant Bacillus spp. in agricultural soil under different management regimes detected via PCR-DGGEMicrobial Ecology45302316CrossRefGoogle Scholar
  16. Martin, N.I., Hu, H., Moake, M.M., Churey, J.J., Whittal, R., Worobo, R.W., Vederas, J.C. 2003Isolation, structural characterization, and properties of mattacin (polymyxin M), a cyclic peptide antibiotic produced by Paenibacillus kobensisJournal of Biological Chemistry2781312413132Google Scholar
  17. Piuri, M., Sanchez-Rivas, C., Ruzal, S.M. 1998A novel antimicrobial activity of a Paenibacillus polymyxa strain isolated from regional fermented sausagesLetters in Applied Microbiology27913CrossRefGoogle Scholar
  18. Ramos, H.J., Roncato-Maccari, L.D., Souza, E.M., Soares-Ramos, J.R., Hungria, M., Pedrosa, F.O. 2002Monitoring Azospirillum–wheat interactions using the gfp and gusA genes constitutively expressed from a new broad-host range vectorJournal of Biotechnology97243252CrossRefGoogle Scholar
  19. Rosado, A.S., Duarte, G.F., Seldin, L. 1994Optimization of electroporation procedure to transform B. polymyxa SCE2 and other nitrogen-fixing BacillusJournal of Microbiological Methods19111CrossRefGoogle Scholar
  20. Rosado, A.S., Seldin, L. 1993Production of a potentially novel anti-microbial substance by Bacillus polymyxaWorld Journal of Microbiology and Biotechnology9521528CrossRefGoogle Scholar
  21. Ross, N., Villemur, R., Marcandella, E., Deschênes, L. 2001Assessment of changes in biodiversity when a community of ultramicrobacteria isolated from groundwater is stimulated to form a biofilmMicrobial Ecology425668Google Scholar
  22. Seldin, L., Elsas, J.D., Penido, E.C.G. 1983Bacillus nitrogen fixers from Brazilian soilsPlant and Soil70243255Google Scholar
  23. Shishido, M., Breuil, C., Chanway, C.P. 1999Endophytic colonization of spruce by plant growth-promoting rhizobacteriaFEMS Microbiology Ecology29191196CrossRefGoogle Scholar
  24. Tombolini, R., Gaag, D.J., Gerhardson, B., Jansson, J.K. 1999Colonization pattern of the biocontrol strain Pseudomonas chlororaphis MA342 on barley seeds visualized by using green fluorescent proteinApplied and Environmental Microbiology6536743680Google Scholar
  25. Tombolini, R., Unge, A., Davey, M.E., Bruijn, F.J., Jansson, J.K. 1997Flow cytometric and microscopic analysis of GFP-tagged Pseudomonas fluorescens bacteriaFEMS Microbiology Ecology221728CrossRefGoogle Scholar
  26. Unge, A., Tombolini, R., Molbak, L., Jansson, J.K. 1999Simultaneous monitoring of cell number and metabolic activity of specific bacterial populations with a dual gfp-luxAB marker systemApplied and Environmental Microbiology65813821Google Scholar
  27. Weid, I., Alviano, D.S., Santos, A.L.S., Soares, R.M.A., Alviano, C.S., Seldin, L. 2003Antimicrobial activity of Paenibacillus peoriae against a broad spectrum of phytopathogenic bacteria and fungiJournal of Applied Microbiology9511431151CrossRefGoogle Scholar
  28. Weid, I., Duarte, G.F., Elsas, J.D., Seldin, L. 2002Paenibacillus brasilensis sp. nov., a novel nitrogen-fixing species isolated from the maize rhizosphere in BrazilInternational Journal of Systematic and Evolutionary Microbiology5221472153Google Scholar
  29. Weid, I., Paiva, E., Nóbrega, A., Elsas, J.D., Seldin, L. 2000Diversity of Paenibacillus polymyxa strains isolated from the rhizosphere of maize planted in cerrado soilResearch in Microbiology151369381Google Scholar
  30. Walker, R., Rossall, S., Asher, M.J.C. 2002Colonization of the developing rhizosphere of sugar beet seedlings by potential biocontrol agents applied as seed treatmentsJournal of Applied Microbiology92228237CrossRefGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • I. von der Weid
    • 1
    • 2
  • V. Artursson
    • 3
  • L. Seldin
    • 2
  • J. K. Jansson
    • 1
    • 3
  1. 1.Section for Natural SciencesSödertörn University CollegeHuddingeSweden
  2. 2.Instituto de Microbiologia Prof. Paulo de Góes, Laboratório de Genética Microbiana, Departamento de Microbiologia Geral, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de JaneiroBrazil
  3. 3.Department of MicrobiologySwedish University for Agricultural SciencesUppsalaSweden

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