Tan spot, caused by Pyrenophora tritici-repentis, is an economically important foliar disease of wheat in Kazakhstan. Population structure of the pathogen changes every year due to climate change. This study aimed to characterize the race structure of P. tritici-repentis isolates recovered from wheat in south and north Kazakhstan, and identify tan spot resistance in host genotypes based on disease phenotypes and molecular screening. Virulence profiles were determined within 40 isolates of the pathogen from wheat crops during the 2020 growing season. Seven races, (1, 3, 4, 5, 6, 7 and 8) were identified. A collection of 80 wheat accessions, including promising lines and cultivars from Kazakhstan and Russia, were evaluated for their reactions to races 1 and 5 of the pathogen, and to Ptr ToxA and Ptr ToxB, using greenhouse assessments and molecular markers diagnostic for the Tsn1 and Tsc2 genes. From a practical viewpoint, 18 wheat genotypes were insensitive to the two races and the two Ptr toxins. This resistant germplasm can be used in breeding programmes aiming to develop wheat varieties resistant to P. tritici-repentis.