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Research on Gene Knockout and Function of FgPDE1 in Fusarium graminearum |
CHANG Yu-mei, HOU Zhan-ming |
College of Life Science and Technology, Inner Mongolia Normal University, Hohhot 010022, China |
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Abstract Objective: The purpose is to knock out FgPDE1 gene in Fusarium graminearum, so as to identify the function of the gene through analysis of the phenotype of the deletion mutants. Methods: The Split-marker strategy is applied to build knockout cassette containing hygromycin phosphotransferase gene and anti-hygromycin transformants are obtained by using PEG-mediated protoplast transformation. The FgPDE1 gene deletion mutants are screened by the absent of the PCR products of the FgPDE1 gene. The function of the gene is analyzed according to the mutant phenotype and pathogenicity detection. Results: The Split-marker knockout cassette is successfully constructed and the transformants are obtained after PEG-mediated transformation of the protoplasts of PH-1 and then, three FgPDE1 gene deletion mutants are obtained through PCR screening. The phenotypic analysis revealed that there is no significant difference between ΔFgPDE1 and wild type in terms of colony phenotype and growth rate. The virulence assay by fruit of tomato infected by conidiospores show that the mutants do not decrease greatly in pathogenicity. However, microscopic observation show that the conidiospore amount of ΔFgPDE1 reduce significantly. Conclusions: FgPDE1 gene might be related to conidia formation of the Fusarium graminearum.
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Received: 10 March 2015
Published: 25 August 2015
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