We assembled an international barley panel comprising 282 entries from 26 countries with various levels of field resistance to leaf rust caused by Puccinia hordei. The panel was screened for leaf rust response with an array of pathotypes at the seedling stage, and at the adult plant stage in multiple environments (2013–2015) in Australia and Uruguay, and genotyped using > 13 K polymorphic DArT-Seq markers. Multipathotype testing in the greenhouse postulated the presence of seedling resistance genes Rph1, Rph2, Rph3, Rph4, Rph7, Rph9.am, Rph12, Rph14, Rph15, Rph19, and Rph25. Genome-wide association studies (GWAS) based on field data identified 13 QTLs significantly associated with DArT-Seq markers on chromosomes 2H (Rph_G_Q1, Rph_G_Q2, Rph_...