Journal article

Agrobacterium -Mediated Transformation of Fusarium oxysporum : An Efficient Tool for Insertional Mutagenesis and Gene Transfer

Mullins E, Chen X, Romaine P, Raina R, Geiser D, Kang S ...see all
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Mullins, E. D., Chen, X., Romaine, P., Raina, R., Geiser, D. M., and Kang, S. 2001. Agrobacterium-mediated transformation of Fusarium oxysporum: An efficient tool for insertional mutagenesis and gene trans-fer. Phytopathology 91:173-180. Agrobacterium tumefaciens-mediated transformation (ATMT) has long been used to transfer genes to a wide variety of plants and has also served as an efficient tool for insertional mutagenesis. In this paper, we report the construction of four novel binary vectors for fungal transfor-mation and the optimization of an ATMT protocol for insertional muta-genesis, which permits an efficient genetic manipulation of Fusarium oxysporum and other phytopathogenic fungi to be achieved. Employing the binary vectors, carrying the bacterial hygromycin B phosphotrans-ferase gene (hph) under the control of the Aspergillus nidulans trpC promoter as a selectable marker, led to the production of 300 to 500 hy-gromycin B resistant transformants per 1 × 10 6 conidia of F. oxysporum, which is at least an order of magnitude higher than that previously accomplished. Transformation efficiency correlated strongly with the duration of cocultivation of fungal spores with Agrobacterium tumefaciens cells and significantly with the number of Agrobacteruium tumefaciens cells present during the cocultivation period (r = 0.996; n = 3; P < 0.01). All transformants tested remained mitotically stable, maintaining their

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  • E D Mullins

  • X Chen

  • P Romaine

  • R Raina

  • D M Geiser

  • S Kang

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