SUMMARY Mitogen-activated protein kinases (MAPKs) play important roles in signal transduction and regulation of various aspects of saprotrophic and pathogenic growth in plant pathogenic fungi. We have generated a Botrytis cinerea knock-out mutant in the bmp3 gene encoding a homologue of the yeast Slt2 cell wall integrity MAPK. The Deltabmp3 mutant showed reduced vegetative growth on various media, strongly impaired conidiation and loss of sclerotia formation. Growth retardation of the mutant was enhanced in media with low osmolarity, whereas nearly wild-type growth rates were observed under high osmolarity conditions. The Deltabmp3 mutant did not show increased susceptibility to cell wall damage induced by glucanase, Calcofluor White or Nikkomycin Z, but was more susceptible to the oxidizing agent paraquat and the phenylpyrrole fungicide fludioxonil. Deltabmp3 conidia showed normal germination and germ tube growth on agar, but excessive germ tube elongation on hard surfaces and reduced penetration efficiency, indicating a defect in surface sensing. After penetration, development of necrotic lesions induced by the Deltabmp3 mutant was retarded. All these defects were restored by genetic complementation of the mutant with the wild-type bmp3 gene.