Skip to main content

Advertisement

Springer Nature Link
Log in

Identification and RAPD-typing of Ewingella americana on cultivated mushrooms in Castilla-La Mancha, Spain

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Ewingella americana was identified as the causal agent of internal stipe necrosis on symptomatic samples collected from mushroom farms located in Castilla-La Mancha, Spain, during the years 2008 to 2010. Six isolates obtained from Agaricus bisporus (button mushroom) and Pleurotus ostreatus (oyster mushroom) were studied to gain insight into their diversity. According to their biochemical profile, the isolates corresponded to biogroup 1. RAPD typing was performed directly with broth cultures of bacteria using two selected primers, and the profiles obtained were used to define four RAPD types. The pathogenicity of isolates was tested by artificial inoculation in A. bisporus and P. ostreatus, in both of which the pathogen reproduced the disease symptoms, and was reisolated from inoculated specimens. Artificial inoculations of the bacteria on Pleurotus eryngii (king oyster mushroom) reproduced the symptoms of the disease, but, until now, this bacterium has not been isolated from this fungus.

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

Access this article

Log in via an institution

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

References

  • Anonymous (2005). http://www2.uni-jena.de/biologie/mikrobio/tipps/rapd.html (accessed on November 9, 2009)

  • Edwards, U., Rogall, T., Blöcker, H., Emde, M., & Böttger, E. C. (1989). Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucl. Acid Research, 17, 7843–7853.

    Article  CAS  Google Scholar 

  • Felsenstein, J. (1989). PHYLIP - phylogeny inference package (version 3.2). Cladistics, 5, 164–166.

    Google Scholar 

  • Fletcher, J. T., & Gaze, R. H. (2008). Mushroom pest and disease control. London: Manson.

    Google Scholar 

  • González, A. J., González-Varela, G., & Gea, F. J. (2009). Brown blotch caused by Pseudomonas tolaasii on cultivated Pleurotus eryngii in Spain. Plant Disease, 93, 667.

    Article  Google Scholar 

  • Goszczynska, T., & Serfontein, J. J. (1998). Milk-tween agar, a semiselective medium for isolation and differentiation of Pseudomonas syringae pv. syringae, Pseudomonas syringae pv. phaseolicola and Xanthomonas axonopodis pv. phaseoli. Journal of Microbiological Methods, 32, 65–72.

    Article  CAS  Google Scholar 

  • Grimont, P. A. D., Farmer, J. J., III, Grimont, F., Asbury, M. A., Brenner, D. J., & Deval, C. (1983). Ewingella americana gen nov., sp nov, a new Enterobacteriaceae isolated from clinical specimens. Annual Review of Microbiology, 134, 39–52.

    Google Scholar 

  • Gwo-Fang, Y., Chiung-Shu, L., & Chann-Chao, C. (1995). Differentiation of Aspergillus parasiticus from Aspergillus sojae by random amplification of polymorphic DNA. Applied and Environmental Microbiology, 61, 2384–2387.

    Google Scholar 

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

    Article  Google Scholar 

  • Hugh, R., & Leifson, E. (1953). The taxonomic significance of fermentative versus oxidative metabolism of carbohydrates by various Gram-negative bacteria. Journal of Bacteriology, 66, 24–26.

    PubMed  CAS  Google Scholar 

  • Inglis, P. W., & Peberdy, J. F. (1996). Isolation of Ewingella americana from the cultivated mushroom, Agaricus bisporus. Current Microbiology, 33, 334–337.

    Article  PubMed  CAS  Google Scholar 

  • Inglis, P. W., Burden, J. L., & Peberdy, J. F. (1996). Evidence for the association of the enteric bacterium Ewingella americana with internal stipe necrosis of Agaricus bisporus. Microbiology, 142, 3253–3260.

    Article  CAS  Google Scholar 

  • Jansing, H., & Rudolph, K. (1990). A sensitivity and quick test for determination of bean seed infestation by Pseudomonas syrinage pv. phaseolicola. Z Pflanzenk Pflanzen, 97, 42–55.

    Google Scholar 

  • King, E. D., Ward, M. K., & Raney, D. E. (1954). Two simple media for the demonstration of pyocyanin and fluorescein. Laboratory and Clinical Medicine, 44, 301–307.

    CAS  Google Scholar 

  • Laconcha, I., López-Molina, N., Rementería, A., Audicana, A., Perales, I., & Garaizar, J. (1998). Phage typing combined with pulsed-field gel electrophoresis and random amplified polymorphic DNA increases discrimination in the epidemiological analysis of Salmonella enteritidis strains. International Journal of Food Microbiology, 40, 27–34.

    Article  PubMed  CAS  Google Scholar 

  • Lee, Ch-J, Jhune, Ch-S, Cheong, J-Ch, Yun, H.-S., & Cho, W.-D. (2009). Occurrence of internal stipe necrosis of cultivated mushrooms (Agaricus bisporus) caused by Ewingella americana in Korea. Mycobiology, 37, 62–66.

    Article  Google Scholar 

  • Lelliott, R. A., & Stead, D. E. (1987). Methods for the diagnosis of bacterial diseases of plants. Oxford: Blackwell Scientific.

    Google Scholar 

  • Lin, A. W., Usera, M. A., Barnet, T. J., & Goldsby, R. A. (1996). Application of random amplified polymorphic DNA analysis to differentiate strains of Salmonella enteritidis. Journal of Clinical Microbiology, 34, 870–876.

    PubMed  CAS  Google Scholar 

  • Miyata, M., Aoki, T., Inglish, V., Yoshida, T., & Endo, M. (1995). RAPD analysis of Aeromonas salmonicida and Aeromonas hydrophila. Journal of Applied Bacteriology, 79, 181–185.

    Article  PubMed  CAS  Google Scholar 

  • Nhung, P. H., Ohkusu, K., Mishima, N., Noda, M., Shah, M. M., Sun, X., Hayashi, M., & Ezaki, T. (2007). Phylogeny and species identification of the family Enterobacteriaceae based on dnaJ sequences. Diagnostic Microbiology and Infectious Disease, 58, 153–161.

    Article  CAS  Google Scholar 

  • Orskov, F., & Orskov, I. (1993). Summary of a workshop on the clone concept. Epidemiology, taxonomy and evolution of the Enterobacteriaceae and other bacteria. Journal of Infectious Diseases, 148, 346–357.

    Article  Google Scholar 

  • Pound, M. W., Tart, S. B., & Okoye, O. (2007). Multidrug-resistant Ewingella americana: a case report and review of the literature. The Annals of Pharmacotherapy, 41, 2066–2070.

    Article  PubMed  Google Scholar 

  • Reyes, J. E., Venturini, M. E., Oria, R., & Blanco, D. (2004). Prevalence of Ewingella americana in retail fresh cultivated mushrooms (Agaricus bisporus, Lentinula edodes and Pleurotus ostreatus) in Zaragoza (Spain). FEMS Microbiology Ecology, 47, 291–296.

    Article  PubMed  CAS  Google Scholar 

  • Roy Chowdhury, P., & Heinemann, J. A. (2006). The general secretory pathway of Burkholderia gladioli pv. agaricicola BG164R is necessary for cavity disease in white button mushrooms. Applied and Environmental Microbiology, 72, 3558–3565.

    Article  PubMed  Google Scholar 

  • Roy Chowdhury, P., Pay, J., & Braithwaite, M. (2007). Isolation, identification and ecology of Ewingella americana (the causal agent of internal stipe necrosis) from cultivated mushrooms in New Zealand. Australasian Plant Pathology, 36, 424–428.

    Article  Google Scholar 

  • Shawkey, M. D., Mills, K. L., Dale, C., & Hill, G. E. (2005). Microbial diversity of wild bird feathers revealed through culture-based and culture-independent techniques. Microbial Ecology, 50, 40–47.

    Article  PubMed  Google Scholar 

  • Smits, T. H., Rezzonico, F., Kamber, T., Blom, J., Goesmann, A., Frey, J. E., & Duffy, B. (2010). Complete genome sequence of the fire blight pathogen Erwinia amylovora CFBP 1430 and comparison to other Erwinia spp. Molecular Plant-Microbe Interactions, 23, 384–393.

    Article  PubMed  CAS  Google Scholar 

  • Soler Rivas, C., Juncà Blanch, G., & Wichers, H. J. (2004). La mancha bacteriana en champiñón y setas. In: Avances en la tecnología de la producción comercial del champiñón y otros hongos cultivados, 2. Patronato de Promoción Económica. Diputación Provincial de Cuenca (ed.), pp. 171–187.

  • Struelens, M. J., et al. (1996). Consensus guidelines for appropiate use and evaluation of microbial epidemiologic typing systems. Clinical Microbiology and Infection, 2, 2–11.

    Article  PubMed  Google Scholar 

  • Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A., & Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acid Research, 18, 6531–6535.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Governments of the Principality of Asturias and Autonomous Community of Castilla-La Mancha. Ana Mª Fernández is a recipient of a grant from the “Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)”.

Author information

Authors and Affiliations

  1. Laboratorio de Fitopatología, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Carretera de Oviedo s/n, 33300, Villaviciosa, Asturias, Spain

    Ana J. González & Ana M. Fernández

  2. Centro de Investigación, Experimentación y Servicios del champiñón (CIES), 16220, Quintanar del Rey, Cuenca, Spain

    Francisco J. Gea & María J. Navarro

Authors
  1. Ana J. González
    View author publications

    Search author on:PubMed Google Scholar

  2. Francisco J. Gea
    View author publications

    Search author on:PubMed Google Scholar

  3. María J. Navarro
    View author publications

    Search author on:PubMed Google Scholar

  4. Ana M. Fernández
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Ana J. González.

Rights and permissions

Reprints and permissions

About this article

Cite this article

González, A.J., Gea, F.J., Navarro, M.J. et al. Identification and RAPD-typing of Ewingella americana on cultivated mushrooms in Castilla-La Mancha, Spain. Eur J Plant Pathol 133, 517–522 (2012). https://doi.org/10.1007/s10658-012-9952-1

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-012-9952-1

Keywords