A cDNA clone, pBH72-F1 (F1), was isolated from a cDNA library prepared from barley leaves 72 h after inoculation with Erysiphe graminis f.sp. hordei. The 1388 bp nucleotide sequence of pBH72-F1 contains an open reading frame encoding a 42.3 kDa polypeptide of 390 amino acids which shows sequence similarity to O-methyltransferases (OMTs) from different plant species; the highest identity (41%) was observed with a putative OMT expressed in roots of maize. A phylogenetic analysis shows that the barley and maize sequences are distinctly different from the ortho-diphenol-OMTs involved in lignin formation. A putative S-adenosyl-L-methionine-binding motif (KELVDDSITN) determined for a rabbit protein-carboxyl OMT is partially conserved in the encoded amino acid sequence. Genomic Southern blot hybridization shows that pBH72-F1 probably represents a single copy gene. The F1 clone corresponds to a gene transcript exhibiting a relatively late accumulation in mildew-infected barley leaves compared to other pathogen-induced transcripts, such as transcripts encoding PR proteins, a peroxidase, and transcripts homologous to a maize caffeic acid OMT. No transcript was detected in plants exhibiting papilla resistance at time points when resistance is thought to be manifested. The atypical transcript accumulation pattern for F1 was also observed after infection by other pathogens and after UV-light treatment.