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Bacillus subtilis M4 decreases plant susceptibility towards fungal pathogens by increasing host resistance associated with differential gene expression

  • Applied Microbial and Cell Physiology
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Abstract

Results presented in this paper describe the ability of Bacillus subtilis strain M4 to reduce disease incidence caused by Colletotrichum lagenarium and Pythium aphanidermatum on cucumber and tomato, respectively. Disease protection in both pathosystems was most probably due to induction of resistance in the host plant since experiments were designed in order to avoid any direct contact between the biocontrol agent and the pathogen. Pre-inoculation with strain M4 thus sensitised both plants to react more efficiently to subsequent pathogen infection. In cucumber, the use of endospores provided a disease control level similar to that obtained with vegetative cells. In contrast, a mixture of lipopeptides from the surfactin, iturin and fengycin families showed no resistance-inducing potential. Interestingly, treatment with strain M4 was also associated with significant changes in gene transcription in the host plant as revealed by cDNA-AFLP analyses. Several AFLP fragments corresponded to genes not expressed in control plants and specifically induced by the Bacillus treatment. In support to the macroscopic protective effect, this differential accumulation of mRNA also illustrates the plant reaction following perception of strain M4, and constitutes one of the very first examples of defence-associated modifications at the transcriptional level elicited by a non-pathogenic bacterium in a host plant.

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References

  • Akpa E, Jacques P, Wathelet B, Paquot M, Fuchs R, Budzikiewicz H, Thonart P (2001) Influence of culture conditions on lipopeptide production by Bacillus subtilis. Appl Biochem Biotechnol 91:551–561

    Article  Google Scholar 

  • Asaka O, Shoda M (1996) Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB14. Appl Environ Microbiol 62:4081–4085

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1993) Phenol/SDS method for plant RNA preparation. In: Current protocols in molecular biology. Wiley, New York, pp 4.3.1–4.3.2

    Google Scholar 

  • Bachem CWB, van der Hoeven RS, de Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF (1996) Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. Plant J 9:745–753

    Article  CAS  PubMed  Google Scholar 

  • Backman PA, Wilson M, Murphy JF (1997) Bacteria for biological control of plant diseases. In: Rechcigl NA, Rechcigl JE (eds) Environmentally safe approaches to crop disease control. Lewis, Boca Raton, pp 95–109

    Google Scholar 

  • Benhamou N, Kloepper J, Quadt-Hallman A, Tuzun S (1996) Induction of defense-related ultrastructural modifications in pea root tissues inoculated with endophytic bacteria. Plant Physiol 112:919–929

    CAS  PubMed  Google Scholar 

  • Brannen PM, Kenney DS (1997) KodiakR—a successful biological-control product for suppression of soil-borne plant pathogens of cotton. J Ind Microbiol Biotechnol 19:169–171

    Article  CAS  Google Scholar 

  • Breyne P, Zabeau M (2001) Genome-wide expression analysis of plant cell cycle modulated genes. Curr Opin Plant Biol 4:136–142

    Article  CAS  PubMed  Google Scholar 

  • Chen TW, Wu WS (1999) Biological control of carrot black rot. J Phytopathol 147:99–104

    Article  Google Scholar 

  • De Torres M, Sanchez P, Fernandez-Delmond I, Grant M (2003) Expression profiling of the host response to bacterial infection: the transition from basal to induced defence responses in RPM1-mediated resistance. Plant J 33:665–676

    Article  PubMed  Google Scholar 

  • Ditt RF, Nester EW, Comai L (2001) Plant gene expression response to Agrobacterium tumefaciens. Proc Natl Acad Sci USA 98:10954–10959

    Article  CAS  PubMed  Google Scholar 

  • Durrant WE, Rowland O, Piedras P, Hammond-Kosack KE, Jones JDG (2000) cDNA-AFLP reveals a striking overlap in race-specific resistance and wound response gene expression profiles. Plant Cell 12:963–977

    Article  CAS  PubMed  Google Scholar 

  • Ferreira JHS, Matthee FN, Thomas AC (1991) Biological control of Eutypa lata on grapevine by an antagonistic strain of Bacillus subtilis. Phytopathology 81:283–287

    Google Scholar 

  • Hammerschmidt R (1999) Induced disease resistance: how do induced plants stop pathogens? Physiol Mol Plant Pathol 55:77–84

    Article  CAS  Google Scholar 

  • Handelsman J, Stabb EV (1996) Biocontrol of soilborne plant pathogens. Plant Cell 8:1855–1869

    Article  CAS  PubMed  Google Scholar 

  • Harris AR, Adkins PG (1999) Versatility of fungal and bacterial isolates for biological control of damping-off disease caused by Rhizoctonia solani and Pythium spp. Biol Control 15:10–18

    Article  Google Scholar 

  • Jacques P, Hbid C, Destain J, Razafindralambo H, Paquot M, De Pauw E, Thonart P (1999) Optimization of biosurfactant lipopeptide production from Bacillus subtilis M4 by Plackett-Burman design. Appl Biochem Biotechnol 77:223–233

    Article  Google Scholar 

  • Kloepper JW, Tuzun S, Kùc JA (1992) Proposed definitions related to induced disease resistance. Biocontrol Sci Technol 2:349–351

    Google Scholar 

  • Leifert C, Li H, Chidburee S, Hampson S, Workman S, Sigee D, Epton HA, Harbour A (1995) Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45. J Appl Bacteriol 78:97–108

    CAS  PubMed  Google Scholar 

  • Maurhofer M, Hase C, Meuwly P, Métraux JP, Defago G (1994) Induction of systemic resistance of tobacco to tobacco necrosis virus by the root-colonizing Pseudomonas fluorescens strain CHA0: influence of the gacA gene and of pyoverdine production. Phytopathology 84:139–146

    CAS  Google Scholar 

  • Métraux JP (2001) Systemic acquired resistance and salicylic acid: current state of knowledge. Eur J Plant Pathol 107:13–18

    Article  Google Scholar 

  • Métraux JP, Burkhart W, Moyer M, Dincher S, Middlesteadt W, Williams S, Payne G, Carnes M, Ryals J (1989) Isolation of a complementary DNA encoding a chitinase with structural homology to a bifunctional lysozyme/chitinase. Proc Natl Acad Sci USA 86:896–900

    PubMed  Google Scholar 

  • Ongena M, Daayf F, Jacques P, Thonart P, Benhamou N, Paulitz TC, Belanger RR (2000) Systemic induction of phytoalexins in cucumber in response to treatments with fluorescent pseudomonads. Plant Pathol 49:523–530

    Article  CAS  Google Scholar 

  • Oostendorp M, Kunz W, Dietrich B, Staub T (2001) Induced disease resistance in plants by chemicals. Eur J Plant Pathol 107:19–28

    Article  CAS  Google Scholar 

  • Pieterse CMJ, Van Pelt JA, Van Wees SCM, Ton J, Leon-Kloosterziel KM, Keurentjes JJB, Verhagen BWM, Knoester M, Van der Sluis I, Bakker P, Van Loon LC (2001) Rhizobacteria-mediated induced systemic resistance: triggering, signalling and expression. Eur J Plant Pathol 107:51–61

    Article  Google Scholar 

  • Raaijmakers JM, Leeman M, van Oorschot MMP, van der Sluis I, Schippers B, Bakker PAHM (1995) Dose-response relationships in biological control of Fusarium wilt of radish by Pseudomonas spp. Phytopathology 85:1075–1081

    Google Scholar 

  • Raupach GS, Kloepper JW (1998) Mixtures of plant growth-promoting rhizobacteria enhance biological control of multiple cucumber pathogens. Phytopathology 88:1158–1164

    Google Scholar 

  • Razafindralambo H, Paquot M, Hbid C, Jacques P, Destain J, Thonart P (1993) Purification of antifungal lipopeptides by reserved-phase high performance liquid chromatography. J Chromatogr 639:81–85

    Article  CAS  PubMed  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Van Loon LC, Van Strien EA (1999) The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant Pathol 55:85–97

    Article  Google Scholar 

  • Van Loon LC, Bakker PAHM, Pieterse CMJ (1998) Systemic resistance induced by rhizosphere bacteria. Annu Rev Phytopathol 36:453–483

    Article  PubMed  Google Scholar 

  • Van Kan JA, Joosten MH, Wagemakers CA, van den Berg-Velthuis GC, de Wit PJ (1992) Differential accumulation of mRNAs encoding extracellular and intracellular PR proteins in tomato induced by virulent and avirulent races of Cladosporium fulvum. Plant Mol Biol 20:513–527

    PubMed  Google Scholar 

  • Wei G, Kloepper JW, Tuzun S (1996) Induced systemic resistance to cucumber diseases and increased plant growth by plant growth-promoting rhizobacteria under field conditions. Phytopathology 86:221–224

    Google Scholar 

  • Whipps JM (2001) Microbial interactions and biocontrol in the rhizosphere. J Exp Bot 52:487–511

    CAS  PubMed  Google Scholar 

  • Yoshida S, Hiradate S, Tsukamoto T, Hatakeda K, Shirata A (2001) Antimicrobial activity of culture filtrate of Bacillus amyloliquefaciens RC-2 isolated from mulberry leaves. Phytopathology 91:181–187

    Google Scholar 

  • Yu GY, Sinclair JB, Hartman GL, Bertagnolli BL (2002) Production of iturin A by Bacillus amyloliquefaciens suppressing Rhizoctonia solani. Soil Biol Biochem 34:955–963

    Article  CAS  Google Scholar 

  • Zehnder GW, Yao CB, Murphy JF, Sikora ER, Kloepper JW (2000) Induction of resistance in tomato against cucumber mosaic cucumovirus by plant growth-promoting rhizobacteria. Biocontrol 45:127–137

    Article  Google Scholar 

  • Zehnder GW, Murphy JF, Sikora EJ, Kloepper JW (2001) Application of rhizobacteria for induced resistance. Eur J Plant Pathol 107:39–50

    Article  Google Scholar 

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Acknowledgements

This work was financed by AGROSTAR s.a. and by a grant from the Walloon Region of Belgium (program AV BIOVAL no 3847). It also received support from the National Funds for Scientific Research (F.N.R.S., Belgium) (Program F.R.F.C. no 2.4.570.00).

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Authors and Affiliations

  1. Centre Wallon de Biologie Industrielle, Faculté Universitaire des Sciences Agronomiques de Gembloux, Université de Liège, Bâtiment B40, Boulevard du Rectorat 29, 4000, Liege, Belgium

    Marc Ongena, Emmanuel Jourdan & Philippe Thonart

  2. Laboratoire de Biologie Moléculaire et Biotechnologie Végétales, Université de Liège, Liege, Belgium

    Francéline Duby & Jacques Dommes

  3. Agrostar, Saint-Servais, Namur, Belgium

    Thierry Beaudry & Victor Jadin

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  1. Marc Ongena
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  2. Francéline Duby
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  3. Emmanuel Jourdan
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  4. Thierry Beaudry
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  5. Victor Jadin
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  6. Jacques Dommes
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  7. Philippe Thonart
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Correspondence to Marc Ongena.

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M. Ongena and F. Duby contributed equally to the paper

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Ongena, M., Duby, F., Jourdan, E. et al. Bacillus subtilis M4 decreases plant susceptibility towards fungal pathogens by increasing host resistance associated with differential gene expression. Appl Microbiol Biotechnol 67, 692–698 (2005). https://doi.org/10.1007/s00253-004-1741-0

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  • DOI: https://doi.org/10.1007/s00253-004-1741-0

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