Summary• R US is a major dominant gene controlling quantitative resistance, inherited from Populus trichocarpa, whereas R 1 is a gene governing qualitative resistance, inherited from P. deltoides.• Here, we report a reiterative process of concomitant fine-scale genetic and physical mapping guided by the P. trichocarpa genome sequence. The highresolution linkage maps were developed using a P. deltoides · P. trichocarpa progeny of 1415 individuals. R US and R 1 were mapped in a peritelomeric region of chromosome 19. Markers closely linked to R US were used to screen a bacterial artificial chromosome (BAC) library constructed from the P. trichocarpa parent, heterozygous at the locus R US .• Two local physical maps were developed, one encompassing the R US allele and the other spanning r US . The alignment of the two haplophysical maps showed structural differences between haplotypes. The genetic and physical maps were anchored to the genome sequence, revealing genome sequence misassembly. Finally, the R US locus was localized within a 0.8-cM interval, whereas R 1 was localized upstream of R US within a 1.1-cM interval.• The alignment of the genetic and physical maps with the local reorder of the chromosome 19 sequence indicated that R US and R 1 belonged to a genomic region rich in nucleotide-binding site leucine-rich repeat (NBS-LRR) and serine threonine kinase (STK) genes.