The STS marker MWG699/TaqI is closely linked to the vrs1 locus and has been proposed as a marker of domestication in barley. This study included 257 cultivated barleys of both two- and six-rowed varieties, mainly from the western Mediterranean region. These included many landraces from the Spanish barley core collection, Moroccan landraces, and a set of accessions from other European countries. Restriction analysis of amplified DNA revealed three alleles, as previously described. Most of the two-rowed entries had the same allele, type K. Six-rowed entries showed both types A and D. Indeed, type D was widespread among Spanish landraces and commercial varieties from central Europe. It was also found in some two-rowed landraces originating from Spain and Morocco. Barleys with the D haplotype were predominantly winter types, whereas the A haplotype was evenly distributed among spring and winter types. These results support the existence of two different genetic sources among six-rowed Spanish landraces.