Puccinia striiformis f. sp. tritici (Pst), the causal pathogen of stripe rust (YR), poses a major threat to global wheat production. The emergence of highly virulent races highlights the need for new sources of genetic resistance in wheat breeding programs. Landraces and historical cultivars preserved in seed banks represent a major source of genetic variation, but relatively unexplored for novel resistances. In this study, we performed a genome- wide association study (GWAS) using 295 bread wheat accessions from the N.I. Vavilov Institute of Plant Genetic Resources (Saint Petersburg, Russia) to identify new sources of YR resistance. The diversity panel was genotyped with DArTseq GBS, and 13,333 polymorphic markers were used for analyses. Disease response was evaluated at the seedling stage in the glass house and adult stage in the field over three years using highly virulent pathotype (134E16 A+Yr17+Yr27), which is prevalent throughout the eastern wheat growing region of Australia. GWAS identified 96 significant marker-trait associations (-log10 P value >3) which represented 67 quantitative trait loci (QTL). Of these 53 QTL were associated with adult plant resistance (APR) and 10 were associated with seedling specific resistance. Moreover, four QTL were associated with all-stage resistance. A total of 12 QTL were deemed novel and were positioned on chromosomes 1D, 3A, 3D, 4A, 4B, 5A, 6B, 7A and 7D. Nine QTL were co-located with cataloged Yr genes while 46 QTL co-located with recently published GWAS studies. These results demonstrate that the Vavilov collection provides a rich source of new alleles which can be used to broaden the genetic base of YR resistance in modern wheat germplasm.