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Simultaneous and selective detection of two major soft rot pathogens of potato: Pectobacterium atrosepticum (Erwinia carotovora subsp. atrosepticum) and Dickeya spp. (Erwinia chrysanthemi)

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Abstract

Dickeya spp. and Pectobacterium atrosepticum are major pathogens of potato. Current methods to detect these soft-rotting bacteria require separate identification steps. Here we describe a simple method allowing simultaneous detection of both pathogens based on multiplex PCR. The sensitivity of the primer sets was first examined on purified genomic DNA of the type strains Dickeya chrysanthemi 2048T and P. atrosepticum 1526T. The specificity and detection limits of the primer sets were successfully tested on 61 strains belonging to various Dickeya and Pectobacterium species, on artificially inoculated and on naturally contaminated potato plants. This new method provides a gain in time and materials, the main advantages for large-scale processes such as pathogen-free seed certification.

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References

  • Alvarez, A. M. (2004). Integrated approaches for detection of plant pathogenic bacteria and diagnosis of bacterial disease. Annual Review of Phytopathology, 42, 339–366.

    Article  PubMed  CAS  Google Scholar 

  • Avrova, A. O., Hyman, L. J., Toth, R. L., & Toth, I. K. (2002). Application of amplified fragment length polymorphism fingerprinting for taxonomy and identification of the soft rot bacteria Erwinia carotovora and Erwinia chrysanthemi. Applied and Environmental Microbiology, 68, 1499–1508.

    Article  PubMed  CAS  Google Scholar 

  • Darrasse, A., Priou, S., Kotoujansky, A., & Bertheau, Y. (1994). PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases. Applied and Environmental Microbiology, 60, 1437–1443.

    PubMed  CAS  Google Scholar 

  • de Boer, S. H., & Ward, L. J. (1995). PCR detection of Erwinia carotovora subsp. atroseptica associated with potato tissue. Phytopathology, 85, 854–858.

    Article  Google Scholar 

  • Fréchon, D., Exbrayat, P., Hélias, V., Hyman, L. J., Jouan, B., Llop, P., et al. (1998). Evaluation of a PCR kit for the detection of Erwinia carotovora subsp. atroseptica on potato tubers. Potato Research, 41, 163–173.

    Article  Google Scholar 

  • Grimberg, J., Maguire, S., & Belluscio, L. (1989). A simple method for the preparation of plasmid and chromosomal E. coli DNA. Nucleic Acid Research, 17, 8893.

    Article  CAS  Google Scholar 

  • Hélias, V., Le Roux, A. C., Bertheau, Y., Andrivon, D., Gauthier, J. P., & Jouan, B. (1998). Characterisation of Erwinia carotovora subspecies and detection of Erwinia carotovora subsp. atroseptica in potato plants, soil and water extracts with PCR-based methods. European Journal of Plant Pathology, 104, 685–699.

    Article  Google Scholar 

  • Hélias, V., Andrivon, D., & Jouan, B. (2000). Internal colonization pathways of potato plants by Erwinia carotovora subsp. atroseptica. Plant Pathology, 49, 33–42.

    Article  Google Scholar 

  • Hyman, L. J., Birch, P. R. J., Dellagi, A., Avrova, A. O., & Toth, I. K. (2000). A competitive PCR-based method for the detection and quantification of Erwinia carotovora subsp. atroseptica on potato tubers. Letters in Applied Microbiology, 30, 330–335.

    Article  PubMed  CAS  Google Scholar 

  • Janse, J. D., & Wenneker, M. (2002). Possibilities of avoidance and control of bacterial plant diseases when using pathogen-tested (certified) or -treated planting material. Plant Pathology, 51, 523–536.

    Article  Google Scholar 

  • Ma, B., Hibbing, M. E., Kil, H. S., Reedy, R. M., Yedidia, I., Breuer, J. J. B., et al. (2007). Host range and molecular phylogenies of the soft rot enterobacterial genera Pectobacterium and Dickeya. Phytopathology, 97, 1150–1663.

    Article  PubMed  Google Scholar 

  • Nassar, A., Darrasse, A., Lemattre, D., Kotoujansky, A., Dervin, C., Vedel, R., et al. (1996). Characterization of Erwinia chrysanthemi by pectinolytic isozyme polymorphism and restriction fragment length polymorphism analysis of PCR-amplified fragments of pel genes. Applied and Environmental Microbiology, 62, 2228–2240.

    PubMed  CAS  Google Scholar 

  • Pérombelon, M. C. M. (1992). Potato blackleg: epidemiology, host-pathogen interaction and control. Netherlands Journal of Plant Pathology, 98, 135–146.

    Article  Google Scholar 

  • Pérombelon, M. C. M. (2002). Potato diseases caused by soft rot erwinias: an overview of pathogenesis. Plant Pathology, 51, 1–12.

    Article  Google Scholar 

  • Singh, U., Trevors, C. M., De Boer, S. H., & Janse, J. D. (2000). Fimbrial-specific monoclonal antibody-based ELISA for European potato strains of Erwinia chrysanthemi and comparison to PCR. Plant Disease, 84, 443–448.

    Article  CAS  Google Scholar 

  • Smadja, B., Latour, X., Faure, D., Chevalier, S., Dessaux, Y., & Orange, N. (2004a). Involvement of N-acylhomoserine lactones throughout plant infection by Erwinia carotovora subsp. atroseptica (Pectobacterium atrosepticum). Molecular Plant Microbe Interaction, 17, 1269–1278.

    Article  CAS  Google Scholar 

  • Smadja, B., Latour, X., Trigui, S., Burini, J.-F., Chevalier, S., & Orange, N. (2004b). Thermodependence of growth and enzymatic activities implicated in pathogenicity of two Erwinia carotovora subspecies (Pectobacterium spp.). Canadian Journal of Microbiology, 50, 19–27.

    Article  CAS  Google Scholar 

  • Smid, E. J., Jansen, A. H. J., & Gorris, L. G. M. (1995). Detection of Erwinia carotovora subsp. atroseptica and Erwinia chrysanthemi in potato tubers using polymerase chain reaction. Plant Pathology, 44, 1058–1069.

    Article  CAS  Google Scholar 

  • Thompson, J. D., Higgins, D. G., & Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acid Research, 22, 4673–4680.

    Article  CAS  Google Scholar 

  • Thomson, S. V., Hildebrand, D. C., & Schroth, M. N. (1981). Identification and differentiation of the Erwinia sugar beet pathogen from members of Erwinia carotovora and Erwinia chrysanthemi. Phytopathology, 71, 1037–1042.

    Article  CAS  Google Scholar 

  • Ward, E., Foster, J. S., Fraaije, B. A., & McCartney, H. A. (2004). Plant pathogen diagnostic: immunological and nucleic acid-based approaches. Annals of Applied Biology, 145, 1–16.

    Article  CAS  Google Scholar 

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Acknowledgments

The authors wish to thank the Plate Forme Technologique d’Evreux, the Region of Haute-Normandie, and the Comité Nord for financial support. We wish to thank V. Hélias, N. Chaabi, A. Chapalain, G. Hemery and M. Vincent for technical assistance.

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Authors and Affiliations

  1. Laboratoire de Microbiologie du Froid, Signaux et Micro-Environnement, UPRES-EA 4312, Centre de Sécurité Sanitaire de Normandie, Université de Rouen, 55 rue Saint-Germain, F-27000, Evreux, France

    Stéphanie Diallo, Xavier Latour, Anne Groboillot, Bruno Smadja, Nicole Orange, Marc G. J. Feuilloley & Sylvie Chevalier

  2. Comité Nord, Station “La Pigache”, Avenue François Mitterand, 62217, Beaurains, France

    Stéphanie Diallo & Patricia Copin

Authors
  1. Stéphanie Diallo
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  2. Xavier Latour
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  3. Anne Groboillot
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  4. Bruno Smadja
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  5. Patricia Copin
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  6. Nicole Orange
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  7. Marc G. J. Feuilloley
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  8. Sylvie Chevalier
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Correspondence to Sylvie Chevalier.

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Diallo, S., Latour, X., Groboillot, A. et al. Simultaneous and selective detection of two major soft rot pathogens of potato: Pectobacterium atrosepticum (Erwinia carotovora subsp. atrosepticum) and Dickeya spp. (Erwinia chrysanthemi). Eur J Plant Pathol 125, 349–354 (2009). https://doi.org/10.1007/s10658-009-9477-4

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  • DOI: https://doi.org/10.1007/s10658-009-9477-4

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