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Inactivation of a candidate effector gene of Zymoseptoria tritici affects its sporulation

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Abstract

Background

Wheat is one of the most important staple crops produced worldwide. Its susceptibility to plant diseases reduces its production significantly. One of the most important diseases of wheat is septoria tritici blotch, a devastating disease observed in fields with wet and temperate conditions. Z. tritici secretes effector proteins to influence the host’s defense mechanisms, as is typical of plant pathogens. In this investigation, we evaluated the pathogenicity of some Zymoseptoria tritici effector candidate genes having a signal peptide for secretion with no known function.

Methods and results

Three genes named Mycgr3G104383, Mycgr3G104444 and Mycgr3G105826 were knocked out separately through homologous recombination, generating Z. tritici IPO323 mutants lacking the functional copy of the corresponding genes. While KO1 and KO3 mutants did not show any significant differences during phenotypic and virulence investigations, the KO2 mutant generated exclusively macropycnidiospores in artificial media, different from wild-type IPO323 which produce only micropycidiospores. The mycelial growth capability of KO2 was also severely attenuated in all of the investigated growth conditions. These changes were observed independent of growth media and growth temperatures, implying that changes were genetic and inherited through generations. Virulence of knockout mutants in wheat leaves was observed to be similar to the wild-type IPO323.

Conclusion

Understanding the biology of Z. tritici and its interactions with wheat will reveal new strategies to fight septoria tritici blotch, enabling breeding wheat cultivars resistant to a broader spectrum of Z. tritici strains. Furthermore, gene knockout via homologous recombination proved to be a powerful tool for discovering novel gene functions.

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Data Availability

All data needed to conduct this study is provided within the manuscript.

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Acknowledgements

Z. tritici IPO323 strain and pCHYG-JK plasmid were kindly provided by Dr. Jason Rudd from Rothamsted Research Institute.

Funding

This project was supported by TÜBİTAK (project number 114O083) under COST action “Pathogen-informed strategies for sustainable broad-spectrum crop resistance”.

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Authors and Affiliations

  1. Department of Plant Production and Technologies, Faculty of Agricultural Science and Technologies, Sivas University of Science and Technology, Sivas, Turkey

    Zemran Mustafa

  2. Department of Plant Protection, Faculty of Agricultural Science and Technologies, Sivas University of Science and Technology, Sivas, Turkey

    Fatih Ölmez

  3. School of Bioengineering, Dalian University of Technology, Dalian, Liaoning, China

    Mahinur Akkaya

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  1. Zemran Mustafa
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  2. Fatih Ölmez
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  3. Mahinur Akkaya
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Contributions

ZM performed knockout experiments and statistical analysis, and wrote the manuscript, FÖ conceptualized and established the methodology. MA Conceptualized and supervised the experiments and reviewed the manuscript.

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Correspondence to Zemran Mustafa.

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Mustafa, Z., Ölmez, F. & Akkaya, M. Inactivation of a candidate effector gene of Zymoseptoria tritici affects its sporulation. Mol Biol Rep 49, 11563–11571 (2022). https://doi.org/10.1007/s11033-022-07879-z

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