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Fusarium graminearum  is an important plant‐pathogenic fungus that causes Fusarium head blight on wheat and barley, and ear rot on maize worldwide. This fungus has been widely used as a model organism to study various biological processes of plant‐pathogenic fungi because of its amenability to genetic manipulation and well‐established outcross system. Gene deletion and overexpression/constitutive expression of target genes are tools widely used to investigate the molecular mechanism underlying fungal development, virulence, and secondary metabolite production. However, for fine‐tuning gene expression and studying essential genes, a conditional gene expression system is necessary that enables repression or induction of gene expression by modifying external conditions. Until now, only a few conditional expression systems have been developed in plant‐pathogenic fungi. This study proposes a new and versatile conditional gene expression system in  F .  graminearum  using the promoter of a copper‐responsive gene, designated  F. graminearum  copper‐responsive 1 ( FCR1 ). Transcript levels of  FCR1  were found to be greatly affected by copper availability conditions. Moreover, the promoter ( P FCR1  ), 1 kb upstream of the  FCR1  open reading frame, was sufficient to confer copper‐dependent gene expression. Replacement of a green fluorescent protein gene and  FgENA5  promoter with a  P FCR1   promoter clearly showed that  P FCR1   could be used for fine‐tuning gene expression in this fungus. We also demonstrated the applicability of this conditional gene expression system to an essential gene study by replacing the promoter of  FgIRE1 , an essential gene of  F .  graminearum . This enabled the generation of  FgIRE1  suppression mutants, which allowed functional characterization of the gene. This study reported the first conditional gene expression system in  F .  graminearum  using both repression and induction. This system would be a convenient way to precisely control gene expression and will be used to determine the biological functions of various genes, including essential ones.

Development of a versatile copper‐responsive gene expression system in the plant‐pathogenic fungus Fusarium graminearum