SUMMARY Botrytis cinerea is a necrotrophic pathogen that produces an array of enzymes capable of attacking the plant cell wall components. We have previously shown that growth of the fungus in planta is accompanied by the degradation of pectin and that endopolygalacturonase (Bcpg) genes are expressed during infection of different plant tissues. It was assumed that pectin demethylation by pectin methylesterases (PME) was essential for the subsequent depolymerization by BcPGs to occur efficiently. We report here on the functional analysis of two Bcpme genes in strain B05.10, using a gene-replacement approach. The method used for the generation of constructs for gene replacement in B. cinerea circumvents the need for cloning and yielded a high proportion of homologous recombinants. Mutants lacking both Bcpme genes are not affected in their growth on highly methylated pectin, nor did they show any reduction in virulence. The results suggest that B. cinerea strain B05.10 can efficiently degrade pectin without prior demethylation.