Summary Globodera pallida and G.'mexicana' are closely related nematode species that can mate and form viable hybrids on tomato but usually develop on different Solanaceous plants. Identification of nematode genes involved in parasitism is important for elucidation of disease resistance mechanisms in plants. In this study, we have used suppression subtractive hybridization (SSH) to investigate differences between the transcriptomes of G. pallida and G.'mexicana' J2s. This provides a basis for further studies characterizing pathogenicity factors in these nematodes. None of the cDNA fragments isolated in the SSH experiments appeared to be completely absent from the other transcriptome. Differences in expression levels of some of the isolated cDNAs between the two species were detected. Sequence analysis revealed that nearly 85% of the cloned sequences are nematode specific and a high proportion were pioneer genes for which no putative homologues were present in the databases. However, homologues of a cellulase and a putative pathogenicity factor previously described from G. rostochiensis were isolated. The putative roles of these sequences in parasitism are discussed.