Zemran Mustafa1, Fatih Ölmez2, Mahinur Akkaya3. 1. Department of Plant Production and Technologies, Faculty of Agricultural Science and Technologies, Sivas University of Science and Technology, Sivas, Turkey. zemran.mustafa@sivas.edu.tr. 2. Department of Plant Protection, Faculty of Agricultural Science and Technologies, Sivas University of Science and Technology, Sivas, Turkey. 3. School of Bioengineering, Dalian University of Technology, Dalian, Liaoning, China.
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.
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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