Abstract
Agrobacterium tumefaciens-mediated transformation (ATMT) is becoming a popular effective system as an insertional mutagenesis tool in filamentous fungi. An efficient Agrobacterium tumefaciens-mediated transformation approach was developed for the plant pathogenic fungus, F. oxysporum, the causal agent of Apple replant disease (ARD) in China. Four parameters were selected to optimize efficiencies of transformation. A. tumefaciens concentration, conidial concentration of F. oxysporum, and co-culture temperature and time have a significant influence on all parameters. Transformants emit green fluorescence under fluorescence microscopy. The integration of a mitotically stable hygromycin resistance gene (hph) in the genome is confirmed by PCR. The transformation efficiency can reach up to 300 transformants per 106 conidia under optimal conditions. This ATMT method is an efficient tool for insertional mutagenesis of F. oxysporum.
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Acknowledgments
We thank Dr. Junxiang Zhang from the Chinese Academy of Agricultural Sciences for kindly providing the Agrobacterium tumefaciens strain LBA4404 and the pCamhybgfp vector. This work was supported partly by the National Key R&D Program of China (2016YFD0201100), the China Agriculture Research System (Cars-27), and the Hebei Natural Science Foundation (c2016204140).
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Dong, YH., Wang, ST. (2022). Agrobacterium tumefaciens-Mediated Transformation Method for Fusarium oxysporum. In: Coleman, J. (eds) Fusarium wilt. Methods in Molecular Biology, vol 2391. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1795-3_6
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DOI: https://doi.org/10.1007/978-1-0716-1795-3_6
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