The rym3 (formerly designated ym3) gene conferring resistance to barley yellow mosaic virus (BaYMV) is effective against all strains of the virus but up to now has not been mapped to any chromosome. We performed a linkage analysis, using DNA extracted from individually harvested mature leaves of 153 F2 plants derived from a cross between BaYMV-resistant cv ’Ishuku Shirazu’ carrying rym3 and susceptible cv ’Ko A’. Additionally, the F3 lines derived from F2 plants were grown in the BaYMV-infested field and examined for their reaction to BaYMV. Our results indicated that rym3 is located on the short arm of chromosome 5H and flanked by RFLP markers MWG28and ABG705A at distances of 7.2 and 11.7 cM, respectively. The chromosomal configuration estimated by DNA markers around rym3 and the utilization of these molecular markers for pyramiding with the BaYMV resistance genes in barley breeding programs are discussed.