Data on plant pathogenic oomycetes are scarce and little is known about the early events leading to the onset of infection. The aim of this work was to analyze the penetration process of the soil-borne plant pathogen Phytophthora parasitica, which has a wide host range. Here, we performed a cytological analysis of the colonization of the first plant cell and developed an inoculation assay for characterizing the entire penetration process through cellular and molecular analyses. We showed that P. parasitica infects roots by producing a specialized structure, the appressorium. We produced the first cDNA library for the penetrating stage of a Phytophthora species and showed it to be highly enriched in pathogenicity-related sequences. These included coding sequences for many cell-degrading enzymes, effectors such as RXLR-containing proteins and proteins involved in protection against plant defense responses. Characterization of the appressorium cDNA library and identification of genes overrepresented early in P. parasitica infection provided us with an unprecedented opportunity to decipher the molecular mechanisms involved in penetration of the plant cells during the initiation of infection by a soil-borne oomycete.