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Background  Powdery mildew diseases are a major phytosanitary issue causing important yield and economic losses in agronomic, horticultural and ornamental crops. Powdery mildew fungi are obligate biotrophic parasites unable to grow on culture media, a fact that has significantly limited their genetic manipulation. In this work, we report a protocol based on the electroporation of fungal conidia, for the transient transformation of  Podosphaera fusca  (synonym  Podosphaera xanthii ), the main causal agent of cucurbit powdery mildew.    Results  To introduce DNA into  P. xanthii  conidia, we applied two square-wave pulses of 1.7 kV for 1 ms with an interval of 5 s. We tested these conditions with several plasmids bearing as selective markers hygromycin B resistance ( hph ), carbendazim resistance ( TUB2 ) or GFP ( gfp ) under control of endogenous regulatory elements from  Aspergillus nidulans ,  Neurospora crassa  or  P. xanthii  to drive their expression. An  in planta  selection procedure using the MBC fungicide carbendazim permitted the propagation of transformants onto zucchini cotyledons and avoided the phytotoxicity associated with hygromycin B.    Conclusion  This is the first report on the transformation of  P. xanthii  and the transformation of powdery mildew fungi using electroporation. Although the transformants are transient, this represents a feasible method for the genetic manipulation of this important group of plant pathogens.

Transient transformation of Podosphaera xanthii by electroporation of conidia