Summary
An improved method for PEG-mediated transformation of the fungal pathogen Fusarium oxysporum was developed which led to at least a 10-fold increase in transformation frequency. This improvement was gained through the analysis of biological factors, viz., protoplast origin, plasmid modifications and protoplast/plasmid ratio. Elecroporation, a recently developed method for introducing DNA into many cell types, was successfully applied. It gave similar transformation efficiencies to those obtained with the chemical method, and thus appeared a valuable alternative. Qualitative features such as integration events were also analysed. Molecular analysis of transformants revealed that a single copy of plasmid DNA was preferentially integrated by electroporation. The respective advantages of the two DNA transfer methods are discussed.
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Abbreviations
- (CM):
-
complete medium (Daboussi 1980)
- (MM):
-
minimal medium (Daboussi 1980)
- (MS):
-
10 mM MOPS, pH 6.3, 1 M sorbitol
- (MSC):
-
MS with 10 mM CaCl2
- (HS):
-
5 mM HEPES, pH 6.3, 20% sucrose
- (PEG):
-
polyethylene glycol
- (μFD):
-
microfarad
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Communicated by B. S. Cox
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Langin, T., Daboussi, M.J., Gerlinger, C. et al. Influence of biological parameters and gene transfer technique on transformation of Fusarium oxysporum . Curr Genet 17, 313–319 (1990). https://doi.org/10.1007/BF00314878
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DOI: https://doi.org/10.1007/BF00314878