Skip to main content
SpringerLink
Log in

The role of antibiosis and induced systemic resistance, mediated by strains of Pseudomonas chlororaphis, Bacillus cereus and B. amyloliquefaciens, in controlling blackleg disease of canola

  • Published:
BioControl Aims and scope Submit manuscript

Abstract

Antibiotic-producing Pseudomonas chlororaphis strains DF190 and PA23, Bacillus cereus strain DFE4 and Bacillus amyloliquefaciens strain DFE16 were tested for elicitation of induced systemic resistance (ISR) and direct antibiosis in control of blackleg in canola caused by the fungal pathogen Leptosphaeria maculans. Inoculation of bacteria 24 h and 48 h prior to the pathogen was crucial for disease control. In systemic induction studies, the bacteria and culture extracts had lower but significant suppression of the blackleg lesion. When inoculated at the same wound site as the pathogen pycnidiospores, the bacterial culture extracts had significantly higher reduction of blackleg lesion development. However, localized plant defense-related enzyme activity at the site of inoculation was not induced by all the bacteria. Direct antifungal activity at the infection site seems to be the dominant mechanism mediating control of L. maculans. A Tn5-gacS mutant of strain PA23 exhibited a complete loss of antifungal and biocontrol activity, which was restored upon addition of the gacS gene in trans. Interestingly, a phenazine-minus derivative of PA23 that produces elevated levels of pyrrolnitrin exhibited the same or higher blackleg disease suppression compared to the wild type. These findings suggest that direct antifungal activity, possibly mediated by pyrrolnitrin, and some low level of induced systemic resistance is involved in P. chlororaphis biocontrol of blackleg.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Audenaert K, Pattery T, Cornelis P, Hofte M (2002) Induction of systemic resistance to Botrytis cinerea in tomato by Pseudomonas aeruginosa 7NSK2: role of salicylic acid, pyochelin, and pyocyanin. Mol Plant Microbe Interact 15:1147–1156

    Article  PubMed  CAS  Google Scholar 

  • Baker KF, Cook RJ (eds) (1974) Biological control of plant pathogens. W.H. Freeman and Company, San Francisco, 433 pp

  • Bakker PAHM, Ran LX, Pieterse CMJ, van Loon LC (2003) Understanding the involvement of rhizobacteria-mediated induction of systemic resistance in biocontrol of plant diseases. Can J Plant Pathol 25:5–9

    Article  Google Scholar 

  • Bargabus RL, Zidack NK, Sherwood JW, Jacobsen BJ (2002) Characterization of systemic resistance in sugar beet elicited by a non-pathogenic, phyllosphere-colonizing Bacillus mycoides, biological control agent. Physiol Mol Plant Pathol 61:289–298

    Article  CAS  Google Scholar 

  • Boller T, Mauch F (1988) Colorimetric assay for chitinase. Methods Enzymol 161:430–435

    Article  CAS  Google Scholar 

  • Buensanteai N, Yue GY, Prathuangwong S (2009) Priming, signaling, and protein production associated with induced resistance by Bacillus amyloliquefaciens KPS46. World J Microbiol Biotechnol 25:1275–1286

    Article  CAS  Google Scholar 

  • Bull CT, Welle DM, Thomashow LS (1991) Relationship between root colonization and suppression of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens strain 2-79. Phytopathology 81:954–959

    Article  Google Scholar 

  • Campbell R (1989) Biological control of microbial plant pathogens. Cambridge University Press, Cambridge

    Book  Google Scholar 

  • Chin-A-Woeng TFC, Bloemberg GV, Mulders IHM, Dekkers LC, Lugtenberg BJJ (2000) Root colonization by phenazine-1-carboxamide-producing bacterium Pseudomonas chlororaphis PCL1391 is essential for biocontrol of tomato foot and root rot. Mol Plant Microbe Interact 12:1340–1345

    Article  Google Scholar 

  • de Boer M, van der Sluis I, van Loon LC, Bakker PAHM (1999) Combining fluorescent Pseudomonas spp. strains to enhance suppression of fusarium wilt of radish. Eur J Plant Pathol 105:201–210

    Article  Google Scholar 

  • De Vleesschauwer D, Djavaheri M, Bakker PAHM, Hofte M (2007) Pseudomonas fluorescens WCS374r-induced systemic resistance in rice against Magnaporthe oryzae is based on pseudobactin-mediated priming for a salicylic acid-repressible multifaceted defense response. Plant Physiol 148:1996–2012

    Article  Google Scholar 

  • Fernando WGD, Nakkeeran S, Zhang Y, Savchuk S (2007) Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals. Crop Prot 26:100–107

    Article  Google Scholar 

  • Hammerschmidt R, Nuckles EM, Kuc J (1982) Association of enhanced peroxidase activity with induced systemic resistance of cucumber to Colletotricum lagenarium. Physiol Mol Pathol 20:73–82

    Article  CAS  Google Scholar 

  • Hammond KE, Lewis BG, Musa TM (1985) A systemic pathway in the infection of oilseed rape plants by Leptosphaeria maculans. Plant Pathol 34:557–565

    Article  Google Scholar 

  • Heeb S, Haas D (2001) Regulatory roles of the GacS/GacA two-component system in plant-associated and other gram-negative bacteria. Mol Plant Microbe Interact 14:1351–1363

    Article  PubMed  CAS  Google Scholar 

  • Iavicoli A, Boutet E, Buchala A, Metraux JP (2003) Induced systemic resistance in Arabidopsis thaliana in response to root inoculation with Pseudomonas fluorescens CHA0. Mol Plant Microbe Interact 16:851–858

    Article  PubMed  CAS  Google Scholar 

  • Jourdan E, Henry G, Duby F, Dommes J, Barthélemy JP, Thonart P, Ongena M (2009) Insights into the defense-related events occurring in plant cells following perception of surfactin-type lipopeptide from Bacillus subtilis. Mol Plant Microbe Interact 22:456–468

    Article  PubMed  CAS  Google Scholar 

  • McCleary BV, Shameer I (1987) Assay of malt beta-glucanase using azo-barley glucan—an improved precipitant. J Inst Brew 93:87–90

    CAS  Google Scholar 

  • Meziane H, Sluis IVD, van Loon LC, Hofte M, Bakker PAHM (2005) Determinants of Pseudomonas putida WCS358 involved in inducing systemic resistance in plants. Mol Plant Pathol 6:177–185

    Article  PubMed  Google Scholar 

  • Olivain C, Alabouvette C, Steinberg C (2004) Production of a mixed inoculum of Fusarium oxysporum Fo47 and Pseudomonas fluorescens C7 to control Fusarium diseases. Biocontrol Sci Technol 14:227–238

    Article  Google Scholar 

  • Ongena M, Jacques P, Touré Y, Destain J, Jabrane A, Thonart P (2005) Involvement of fengycin-type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis. Appl Microbiol Biotechnol 69:29–38

    Article  PubMed  CAS  Google Scholar 

  • Ongena M, Jourdan E, Adam A, Paquot M, Bran A, Joris B, Arpigny JL, Thonart P (2007) Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants. Environ Microbiol 9:1084–1090

    Article  PubMed  CAS  Google Scholar 

  • Pang Y, Liu X, Ma Y, Chernin L, Berg G, Gao K (2009) Induction of systemic resistance, root colonization and biocontrol activities of the rhizospheric strain of Serratia plymuthica are dependent on N-acyl homoserine lactones. Eur J Plant Pathol 124:261–268

    Article  CAS  Google Scholar 

  • Parke JL (1990) Population dynamics of Pseudomonas cepacia in the pea spermosphere in relation to biocontrol of Pythium. Phytopathology 80:1307–1311

    Article  Google Scholar 

  • Pieterse CMJ, van Pelt JA, van Wees SCC, Ton J, Kloosterzie KML, Keurentjes JJB, Verhagen VWM, Knoester M, Sluis IVD, Bakker PAHM, van Loon LC (2001) Rhizobacteria-mediated induced systemic resistance: triggering, signaling and expression. Eur J Plant Pathol 107:51–61

    Article  Google Scholar 

  • Poritsanos NJ (2005) Molecular mechanisms involved in the secondary metabolite production and biocontrol of Pseudomonas chlororaphis PA23. Masters Thesis Dissertation, University of Manitoba

  • Poritsanos N, Selin C, Fernando WGD, Nakkeeran S, de Kievit TR (2006) A GacS deficiency does not affect Pseudomonas chlororaphis PA23 fitness when growing on canola, in aged batch culture or as a biofilm. Can J Microbiol 52:1177–1188

    Article  PubMed  CAS  Google Scholar 

  • Ramarathnam R (2007) Mechanisms of phyllosphere biological control of Leptosphaeria maculans, the blackleg pathogen of canola, using antagonistic bacteria. PhD Thesis, University of Manitoba

  • Ramarathnam R, Fernando WGD (2006) Preliminary phenotypic and molecular screening for potential bacterial biocontrol agents of Leptosphaeria maculans, the blackleg pathogen of canola. Biocontrol Sci Technol 16:567–582

    Article  Google Scholar 

  • Ran LX, Li ZN, Wu GJ, van Loon LC, Bakker PAHM (2005) Induction of systemic resistance against bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. Eur J Plant Pathol 113:59–70

    Article  CAS  Google Scholar 

  • Selin C, Habibian R, Poritsanos N, Athukorala SNP, Fernando D, De Kievit TR (2010) Phenazines are not essential for Pseudomonas chlororaphis PA23 biocontrol of Sclerotinia sclerotiorum, but do play a role in biofilm formation. FEMS Microbiol Ecol 71:73–83

    Article  PubMed  CAS  Google Scholar 

  • Thomashow LS, Bonsall RF, Weller DM (1997) Antibiotic production by soil and rhizosphere microbes in situ. In: Hurst CJ, Knusden GR, McInerney MJ, Stetzenbach LD, Walter MD (eds) Manual of environmental microbiology. ASM Press, Washington, DC, pp 493–499

    Google Scholar 

  • Tran H, Ficke A, Asiimwe T, Höfte M, Raaijmakers JM (2007) Role of the cyclic lipopeptide massetolide A in biological control of Phytophthora infestans and in colonization of tomato plants by Pseudomonas fluorescens. New Phytol 175:731–742

    Article  PubMed  CAS  Google Scholar 

  • Weller DM, Howie WJ, Cook RJ (1988) Relationship between in vitro inhibition of Gaeumannomyces graminis var tritici and suppression of take-all of wheat by fluorescent pseudomonads. Phytopathology 78:1094–1100

    Article  Google Scholar 

  • Zhang Y, Fernando WGD, de Kievit T, Berry C, Daayf F, Paulitz T (2006) Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction. Can J Microbiol 52:476–481

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the financial support for this work through grants awarded to W.G.D.F. and T.R.de K. from the Natural Sciences and Engineering Research Council (NSERC) Discovery Grants Program.

Author information

Author notes

  1. Rajesh Ramarathnam

    Present address: Canadian Food Inspection Agency, 59 Camelot Drive, Ottawa, ON, KIA 0Y9, Canada

Authors and Affiliations

  1. Department of Plant Science, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Rajesh Ramarathnam & W. G. Dilantha Fernando

  2. Department of Microbiology, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Teresa de Kievit

Authors
  1. Rajesh Ramarathnam
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W. G. Dilantha Fernando
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Teresa de Kievit
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to W. G. Dilantha Fernando.

Additional information

Handling Editor: Monica Hofte.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ramarathnam, R., Fernando, W.G.D. & de Kievit, T. The role of antibiosis and induced systemic resistance, mediated by strains of Pseudomonas chlororaphis, Bacillus cereus and B. amyloliquefaciens, in controlling blackleg disease of canola. BioControl 56, 225–235 (2011). https://doi.org/10.1007/s10526-010-9324-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10526-010-9324-8

Keywords

Search

Navigation