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Improved recovery of Erwinia amylovora-stressed cells from pome fruit on RESC, a simple, rapid and differential medium

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Abstract

The bacterium Erwinia amylovora causes fire blight, a serious and widespread disease of several pome fruit and ornamental plants. The use of suitable detection tools is essential for preventing its dissemination and, according to the protocol of the European and Mediterranean Plant Protection Organization, the isolation and further identification of E. amylovora is the only conclusive test of its presence. However, bacterial growth on solid media can be hampered when the pathogen is suffering stressful conditions in pome fruit or in other habitats. Since copper is an essential micronutrient that, in E. amylovora, also increases the exopolysaccharide production in rich-nutrient media, we have designed a non-selective differential medium containing 1.5 mM CuSO4 to improve the recovery of E. amylovora from plants under unfavorable conditions. In this new medium named Recovery Erwinia amylovora-Stressed Cells (RESC), its colonies were easily distinguished by a light yellow color and a high mucus production. The plating recovery of several E. amylovora strains in vitro and from naturally infected samples was significantly improved with respect to other media routinely employed, particularly when the pathogen was suffering stressful conditions. Thus, the recovery of stressed E. amylovora cells (after UV irradiation, nutrient deprivation, or the presence of copper ions in non-copper-complexing media) was significantly enhanced on RESC medium, and their culturability period extended. Therefore, RESC is a useful and valuable medium for the isolation of E. amylovora when adverse conditions in the natural environment are expected.

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Acknowledgments

This work was supported through projects AGL2005-06982 and AGL2008-05723-C02-01 from the Ministerio de Educación y Ciencia (MEC) of Spain and GV-05/214 from Generalitat Valenciana, and was performed in the framework of COST 864. M. Ordax thanks a predoctoral fellowship from the MEC, Spain, and E. Marco-Noales has a contract from the Spanish Ministry of Education and Science (Programa INIA-CCAA) co-funded by European Social Fund. The authors wish to specially thank J.M Quesada and M. T. Gorris (IVIA, Spain) for their PCR and ELISA analysis, respectively, E. Carbonell and J. Pérez (IVIA, Spain) for statistical analysis, and J. Murillo (UPNA, Spain) for useful advice. We also thank K. Geider (JKI, Germany) and E. Montesinos (INTEA, Spain) for the bacterial strains provided, and S. Rubio and M. Marín (Servicio de Laboratorios Agroalimentarios de La Rioja, Spain) for permitting us the sampling from orchards with fire blight symptoms.

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  1. Ester Marco-Noales

    Present address: Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera, Km 4.5, 46113, Moncada, Valencia, Spain

Authors and Affiliations

  1. Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera, Km 4.5, 46113, Moncada, Valencia, Spain

    Mónica Ordax & María M. López

  2. Departamento de Microbiología y Ecología, Universidad de Valencia, Av. Dr. Moliner, 50, 46100, Burjassot, Valencia, Spain

    Elena G. Biosca

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  1. Mónica Ordax
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  2. Elena G. Biosca
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  3. María M. López
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  4. Ester Marco-Noales
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Correspondence to Ester Marco-Noales.

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Communicated by D. Treutter.

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Ordax, M., Biosca, E.G., López, M.M. et al. Improved recovery of Erwinia amylovora-stressed cells from pome fruit on RESC, a simple, rapid and differential medium. Trees 26, 83–93 (2012). https://doi.org/10.1007/s00468-011-0614-2

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