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Identification and characterization of Phoma tracheiphila mutants impaired in pathogenicity following Agrobacterium-mediated mutagenesis

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Abstract

Mal secco disease, caused by the pathogenic fungus Phoma tracheiphila, is a devastating disease of susceptible citrus species, especially lemon. To study the molecular interactions between the pathogen and its host, a method for identifying the genes involved in pathogenicity is needed. This work describes the transformation of P. tracheiphila phialoconidia by Agrobacterium tumefaciens, and the generation of mutated P. tracheiphila isolates exhibiting reduced virulence on rough lemon seedlings. A rapid, replicable, and reliable method for screening P. tracheiphila mutants to assess their virulence by using rough lemon seedlings was developed. Among 2263 transformants obtained, three were non-virulent and 43 displayed reduced virulence. In addition, one of the transformants, which exhibited virulence similar to that of the wild type, was used for in planta visualization of the fungus progression through the plant xylem. To our knowledge, this is the first report of A. tumefaciens-mediated transformation of P. tracheiphila, and subsequent screening of the transformants to identify non-virulent mutants.

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Acknowledgments

This work was partially financed by the Chief Scientist of the Israel Ministry of Agriculture and Rural Development (MOARD), research project number 132-1243-06. Contribution No. 523/13 from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel.

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Authors and Affiliations

  1. Department of Plant Pathology, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel

    Tammy Kroitor-Keren, Orna Liarzi, Tami Gat, Julia Skovorodnikova & David Ezra

  2. Plant Sciences Institute, Ornamental Horticulture, Microscopy Unit, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel

    Eduard Belausov

Authors
  1. Tammy Kroitor-Keren
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  2. Orna Liarzi
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  3. Tami Gat
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  4. Julia Skovorodnikova
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  5. Eduard Belausov
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  6. David Ezra
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Correspondence to David Ezra.

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Kroitor-Keren, T., Liarzi, O., Gat, T. et al. Identification and characterization of Phoma tracheiphila mutants impaired in pathogenicity following Agrobacterium-mediated mutagenesis. Phytoparasitica 41, 491–502 (2013). https://doi.org/10.1007/s12600-013-0328-7

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  • DOI: https://doi.org/10.1007/s12600-013-0328-7

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