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Application of the Cre/lox System to Construct Auxotrophic Markers for Quantitative Genetic Analyses in Fusarium graminearum

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Book cover Plant Pathogenic Fungi and Oomycetes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1848))

Abstract

The bacteriophage P1 Cre/lox system has been utilized in diverse fungi for marker recycling and exchange, generation of targeted chromosome translocations, and targeted deletion of interstitial chromosome segments. Here we show the application of this tool in the wheat and maize pathogen, Fusarium graminearum. We explored three different ways to introduce Cre into strains with floxed genes, namely transformation with an episomal or integrative plasmid (pLC28), fusion of protoplasts of strains carrying floxed genes with strains expressing Cre by forcing heterokaryons, and crosses between strains with floxed genes and strains expressing Cre to isolate progeny in which the target genes had been deleted during the cross. We used this system for the construction of strains bearing auxotrophic markers that were generated by gene replacement with positively selectable markers followed by Cre-mediated marker excision. In addition, updated protocols for transformation and crosses for F. graminearum are provided. In combination, strains and tools developed here add to the arsenal of methods that can be used to carry out molecular genetics with F. graminearum.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA

    Lanelle R. Connolly, Allyson A. Erlendson, Corinne M. Fargo, Kendra K. Jackson, Morgan M. G. Pelker, Jacob W. Mazzola, Mark S. Geisler & Michael Freitag

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  1. Lanelle R. Connolly
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  2. Allyson A. Erlendson
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  3. Corinne M. Fargo
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  4. Kendra K. Jackson
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  5. Morgan M. G. Pelker
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  6. Jacob W. Mazzola
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  7. Mark S. Geisler
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  8. Michael Freitag
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Correspondence to Michael Freitag .

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Editors and Affiliations

  1. Department of Microbiology and Plant Pathology, University of California Riverside, Riverside, CA, USA

    Wenbo Ma

  2. Oregon State University, Corvallis, OR, USA

    Thomas Wolpert

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Connolly, L.R. et al. (2018). Application of the Cre/lox System to Construct Auxotrophic Markers for Quantitative Genetic Analyses in Fusarium graminearum. In: Ma, W., Wolpert, T. (eds) Plant Pathogenic Fungi and Oomycetes. Methods in Molecular Biology, vol 1848. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8724-5_16

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  • DOI: https://doi.org/10.1007/978-1-4939-8724-5_16

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