A clone encoding a putative soluble epoxide hydrolase (EH-1), an enzyme which converts epoxides to diols, was isolated by differential screening of a cDNA library prepared from tobacco mosaic virus (TMV)-infected tobacco leaves. To confirm that EH-1 encodes an epoxide hydrolase, the recombinant EH-1 protein produced in bacteria was shown to have high epoxide hydrolase activity in vitro. Infection of resistant but not susceptible tobacco cultivars induced the accumulation of EH-1 transcripts in both the inoculated and uninoculated, systemic leaves. EH-1 expression was also induced in the inoculated and systemic tissues of TMV-infected NahG plants, which are unable to accumulate salicylic acid (SA). However, EH-1 expression in the inoculated leaves of NahG plants was delayed, whilst in the systemic leaves the induction was both later and weaker, compared to that observed in wild-type plants. Furthermore, exogenously applied SA or its functional analog 2,6-dichloroisonicotinic acid (INA) caused a rapid and transient accumulation of EH-1 transcripts, whereas an inactive SA analog did not. Thus, the induction of EH-1 gene expression appears to be regulated by both SA-independent and SA-dependent pathways. Since EH-1 was expressed only in TMV-resistant tobacco after infection, and the encoded enzyme is thought to help metabolize toxic compounds, we propose that EH-1 may play a role in protection from oxidative damage associated with defense responses. It may also play a role in generating signals for activation of certain defense responses.