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Infection and colonization of strawberry by Gnomonia fragariae strain expressing green fluorescent protein

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Abstract

Gnomonia fragariae is a poorly studied ascomycete, which was recently demonstrated to be a cause of severe root rot and petiole blight of strawberry. The pathogen was genetically transformed with the GFP as a vital marker and hygromycin resistance gene. Several stable transformants were obtained, which did not differ in their phenotype from the wild type isolate. Using one of the GFP-tagged isolates the infection process and colonization of roots and petioles of host plant by the pathogen were studied. Fluorescence microscopy examinations of the inoculated plants at different time points showed that plant infection occurs 24 h after inoculation and intensively continues during first 3 days. The specific penetration sites on epidermal cells and preferences in colonization for certain root and petiole tissues were observed. The pathogen intensively colonized and destroyed cortex of roots and petioles and spread rapidly longitudinally within intercellular spaces. The petioles were colonized by the hyphae, which grew mostly in the intracellular spaces of the cortical cells while in the roots the intracellular growth of hyphae occurred only in the later stages of infection. The fungus was also capable to infect the vascular tissues of petioles although these were not the primary tissues colonized by the pathogen. The mature ascomata were formed on the infected petiole bases several weeks after the inoculation. This study presents a genetic transformation method for Gnomonia fragariae and it demonstrates details on infection process and colonization of root, crown and petiole tissues of strawberry by the pathogen.

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Acknowledgements

The research was financially supported by the Swedish Institute and Strategic Foundation for Environmental Research (MISTRA). We are grateful to B. Gillian Turgeon, Cornell University, USA for kindly providing plasmid pIGPAPA and to Lynda M. Ciuffetti, Oregon State University, USA for providing plasmid pCT74. The preparation of the manuscript was supported by the grant 2009/0228/1DP/1.1.1.2.0/09/APIA/VIAA/035

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Author notes

  1. Inga Moročko-Bičevska

    Present address: Latvia State Institute of Fruit-Growing, Graudu str. 1, LV-3701, Dobele, Latvia

Authors and Affiliations

  1. MASE Laboratories AB, Box 148, 751 04, Uppsala, Sweden

    Inga Moročko-Bičevska & Jamshid Fatehi

  2. Department of Forest Mycology and Pathology, Swedish University of Agricultural Sciences, Box 7026, 750 07, Uppsala, Sweden

    Inga Moročko-Bičevska

Authors
  1. Inga Moročko-Bičevska
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  2. Jamshid Fatehi
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Correspondence to Inga Moročko-Bičevska.

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Moročko-Bičevska, I., Fatehi, J. Infection and colonization of strawberry by Gnomonia fragariae strain expressing green fluorescent protein. Eur J Plant Pathol 129, 567–577 (2011). https://doi.org/10.1007/s10658-010-9720-z

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