Phylogenesis of virulent races of Puccinia graminis f. sp. tritici based on phenotyping and genotyping using newly developed SNP markers for genetic groups in clades

Ug99 (TTKSK) and Digalu (TKTTF) race groups of Puccinia graminis f. sp. tritici (Pgt), are thought to be the most virulent, rendering many wheat stem rust resistance genes (Sr) ineffective. A total of 600 single pustule Pgt isolates were collected from eight Egyptian provinces during 2016–2022, undergoing both phenotyping and genotyping assays. Among these, 129 viable samples (v-samples) underwent phenotyping using the North American differentials delineated to Pgt races in the TTKS (Ug99) and TKTTF race groups, identifying 24 distinct physiologic races of Pgt. Notably, seven virulent races, TKTTF (Digalu), TKKTF, TKPTF, TKKTP, TTRTF, TTTTF, and TKFTF, were discovered in Egypt. The host reaction of wheat germplasm to these seven races indicated a high level of susceptibility. A set of 437 dead samples (d-samples) of Pgt isolates from Egypt were genotyped using newly created 17 core SNP markers. Genotypic analysis revealed that these seven races could be classified into two major clades: III and IV. Within these clades, six subclades were identified, namely III-B, IV-A.2, IV-B, IV-E.1, IV-E.2, and IV-F. These SNP genotypes exhibited a broad geographic distribution, highlighting the global movement of Pgt populations across Europe, Asia, and Africa. Notably, none of these genetic groups were present in Egypt before 2016. The most prevalent genotypes were subclades IV-A.2 (TKTTF; Digalu), IV-E.2 (TKKTF, TKPTF, TKFTF), and III-B (TTRTF). The prevalence of these unique genotypes/races suggests clonal reproduction and evolution within them. Race TKKTP (subclade IV-E.1) is particularly concerning because of its ability to overcome the resistance conferred by gene Sr24 against race TTKSK (Ug99). Phylogenetic analysis of Pgt genetic groups revealed that the majority of identified races belong to TKTTF (Digalu) race group (subclade IV-A.2). Notably, no races were found to belong to the TTKS (Ug99) race group (clade I). This information should help breeders select a broader range of resistance genes that are both diverse and effective to enhance the durability of stem rust resistance. •Race phenotyping resulted in identification of 24 races of Puccinia graminis f. sp. tritici (pgt).•Seven Pgt races revealed a broad distribution, virulence and diverse range.•Genotyping using a new custom SNP assay divided Pgt races into two major clades, III and IV.•Genotyping/races belonged to Digalu race (subclade IV-A.2), but not to Ug99 race (clade I).