KEY MESSAGE:Stripe rust resistance gene, Yr82, was mapped in chromosome 3BL using SNP markers. Yr82 interacted with Yr29 to produce lower stripe rust responses at the adult plant stage. Landrace Aus27969 produced low infection types against Australian Puccinia striiformis f. sp. tritici (Pst) pathotypes. A recombinant inbred line (RIL) F7 population from the Aus27969/Avocet S cross was developed. Monogenic segregation for seedling stripe rust response was observed among the RIL population, and the resistance locus was named Yr82. Bulk segregant analysis performed using the iSelect wheat 90 K Infinium SNP array located Yr82 in the long arm of chromosome 3B. The RIL population was screened against stripe rust under field conditions and was genotyped with targeted genotyping-by-sequencing assay. QTL analysis detected the involvement of chromosomes 1B and 3B in controlling stripe rust resistance carried by Aus27969. Incorporation of Yr82 and marker SNPLr46G22 into the linkage map showed that the QTL in 1B and 3B represented Yr29 and Yr82, respectively. Kompetitive allele-specific PCR (KASP) markers sun KASP_300 and KASP_8775 flanked Yr82 distally and proximally, respectively, each at 2 cM distance. These Yr82-linked markers were polymorphic among 84% of Australian cultivars and can be used for marker-assisted selection of Yr82.