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Multiple knockout mutants reveal a high redundancy of phytotoxic compounds that determine necrotrophic pathogenesis of Botrytis cinerea

Thomas Leisen, Janina Werner, Patrick Pattar, Edita Ymeri, Frederik Sommer, Michael Schroda, David Scheuring, Matthias Hahn
doi: https://doi.org/10.1101/2021.08.21.457223
Thomas Leisen
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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Janina Werner
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
2Botanical Institute and Cluster of Excellence on Plant Sciences (CEPLAS), BioCenter, University of Cologne, Zülpicher Str. 47a, Cologne, 50674, Germany
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Patrick Pattar
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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Edita Ymeri
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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Frederik Sommer
3Department of Biology, Molecular Biotechnology & Systems Biology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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Michael Schroda
3Department of Biology, Molecular Biotechnology & Systems Biology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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David Scheuring
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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Matthias Hahn
1Department of Biology, Phytopathology group, Technische Universität Kaiserslautern, Paul-Ehrlich Straße 22, D-67663 Kaiserslautern, Germany
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  • For correspondence: hahn@biologie.uni-kl.de
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Abstract

Botrytis cinerea is a major pathogen of more than 1400 plant species. During infection, the kills host cells during infection and spreads through necrotic tissue, which is believed to be supported by induction of programmed plant cell death. To comprehensively evaluate the contributions of most of the currently known plant cell death inducing proteins (CDIPs) and metabolites for necrotrophic infection, an optimized CRISPR/Cas protocol was established which allowed serial marker-free mutagenesis to generate Botrytis mutants lacking up to 12 different CDIPs. Infection analysis revealed a decrease in virulence with increasing numbers of knockouts, and differences in the effects of knockouts on different host plants. The on planta secretomes obtained from these mutants revealed substantial remaining necrotic activity after infiltration into leaves. Our study has addressed for the first time the functional redundancy of virulence factors of a fungal pathogen, and demonstrates that B. cinerea releases a highly redundant cocktail of proteins and metabolites to achieve necrotrophic infection of a wide variety of host plants.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted August 22, 2021.
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Multiple knockout mutants reveal a high redundancy of phytotoxic compounds that determine necrotrophic pathogenesis of Botrytis cinerea
Thomas Leisen, Janina Werner, Patrick Pattar, Edita Ymeri, Frederik Sommer, Michael Schroda, David Scheuring, Matthias Hahn
bioRxiv 2021.08.21.457223; doi: https://doi.org/10.1101/2021.08.21.457223
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Multiple knockout mutants reveal a high redundancy of phytotoxic compounds that determine necrotrophic pathogenesis of Botrytis cinerea
Thomas Leisen, Janina Werner, Patrick Pattar, Edita Ymeri, Frederik Sommer, Michael Schroda, David Scheuring, Matthias Hahn
bioRxiv 2021.08.21.457223; doi: https://doi.org/10.1101/2021.08.21.457223

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