Abstract
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)2Cys6 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: [Figure not available: see fulltext.]
Original language | English |
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Article number | 25 |
Journal | Fungal Biology and Biotechnology |
Volume | 6 |
Issue number | 1 |
ISSN | 2054-3085 |
DOIs | |
Publication status | Published - 2019 |
Bibliographical note
© The Author(s) 2019.Keywords
- Agrobacterium tumefaciens-mediated transformation
- Bostrycoidin
- Fluorescence
- Fusarium
- Fusarubin
- Heterologous expression
- Polyketides
- Secondary metabolites
- Transformation
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MOESM8 of A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani
Nielsen, M. R. (Creator), Holzwarth, A. K. R. (Creator), Brew, E. (Creator), Chrapkova, N. (Creator), Kaniki, S. E. K. (Contributor), Kastaniegaard, K. (Creator), Sørensen, T. (Creator), Westphal, K. R. (Creator), Wimmer, R. (Creator), Søndergaard, T. (Creator) & Sørensen, J. L. (Creator), Figshare, 2019
DOI: 10.6084/m9.figshare.11352794.v1, https://doi.org/10.6084%2Fm9.figshare.11352794.v1
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MOESM1 of A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani
Nielsen, M. R. (Creator), Holzwarth, A. K. R. (Creator), Brew, E. (Creator), Chrapkova, N. (Creator), Kaniki, S. E. K. (Contributor), Kastaniegaard, K. (Creator), Sørensen, T. (Creator), Westphal, K. R. (Creator), Wimmer, R. (Creator), Søndergaard, T. (Creator) & Sørensen, J. L. (Creator), Figshare, 2019
DOI: 10.6084/m9.figshare.11352743.v1, https://doi.org/10.6084%2Fm9.figshare.11352743.v1
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MOESM3 of A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani
Nielsen, M. R. (Creator), Holzwarth, A. K. R. (Creator), Brew, E. (Creator), Chrapkova, N. (Creator), Kaniki, S. E. K. (Contributor), Kastaniegaard, K. (Creator), Sørensen, T. (Creator), Westphal, K. R. (Creator), Wimmer, R. (Creator), Søndergaard, T. (Creator) & Sørensen, J. L. (Creator), Figshare, 2019
DOI: 10.6084/m9.figshare.11352758.v1, https://doi.org/10.6084%2Fm9.figshare.11352758.v1
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