Ectomycorrhiza development involves the differentiation of structurally specialized fungal tissues (e.g., mantle and Hartig net) and an interface between symbionts. Polypeptides presenting a preferential, up-, or down-regulated synthesis have been characterized in several developing ectomycorrhizal associations. Their spatial and temporal expressions have been characterized by cell fractionation, two-dimensional polyacrylamide gel electrophoresis, and immunochemical assays in the Eucalyptus spp. – Pisolithus tinctorius mycorrhizas. These studies have emphasized the importance of fungal cell wall polypeptides during the early stages of the ectomycorrhizal interaction. The increased synthesis of 30- to 32-kDa acidic polypeptides, together with the decreased accumulation of a prominent 95-kDa mannoprotein provided evidence for major alterations of Pisolithus tinctorius cell walls during mycorrhiza formation. Differential cDNA library screening and shotgun cDNA sequencing were used to clone symbiosis-regulated fungal genes. Several abundant transcripts showed a significant amino acid sequence similarity to a family of secreted morphogenetic fungal proteins, the so-called hydrophobic. In P. tinctorius, the content of hydrophobin transcripts is high in aerial hyphae and during the ectomycorrhizal sheath formation. Alteration of cell walls and the extracellular matrix is therefore a key event in the ectomycorrhiza development. An understanding of the molecular mechanisms that underlies the temporal and spatial control of genes and proteins involved in the development of the symbiotic interface is now within reach, as more sophisticated techniques of molecular and genetic analysis are applied to the mycorrhizal interactions.