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Background  Besides their ability to produce several interesting bioactive secondary metabolites, members of the  Fusarium solani  species complex comprise important pathogens of plants and humans. One of the major obstacles in understanding the biology of this species complex is the lack of efficient molecular tools for genetic manipulation.    Results  To remove this obstacle we here report the development of a reliable system where the vectors are generated through yeast recombinational cloning and inserted into a specific site in  F. solani  through  Agrobacterium tumefaciens -mediated transformation. As proof-of-concept, the enhanced yellow fluorescent protein (eYFP) was inserted in a non-coding genomic position of  F. solani  and subsequent analyses showed that the resulting transformants were fluorescent on all tested media. In addition, we cloned and overexpressed the Zn(II) 2 Cys 6  transcriptional factor  fsr6  controlling mycelial pigmentation. A transformant displayed deep red/purple pigmentation stemming from bostrycoidin and javanicin.    Conclusion  By creating streamlined plasmid construction and fungal transformation systems, we are now able to express genes in the crop pathogen  F. solani  in a reliable and fast manner. As a case study, we targeted and activated the fusarubin ( PKS3 :  fsr ) gene cluster, which is the first case study of secondary metabolites being directly associated with the responsible gene cluster in  F. solani  via targeted activation. The system provides an approach that in the future can be used by the community to understand the biochemistry and genetics of the  Fusarium solani  species complex, and is obtainable from Addgene catalog #133094.    Graphic abstract

A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani