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Leaf rust, caused by Puccinia triticina (Pt), is a major disease of wheat and a significant problem for wheat production in Brazil. The Brazilian variety Toropi, released in 1965, has maintained high levels of field, adult plant resistance (APR) to leaf rust across global locations, while microscopic studies have indicated prehaustorial resistance mechanisms. Analyses of gene expression in flag leaves of Toropi, during the early stages of Pt infection, were undertaken to explore the mechanisms behind the APR in Toropi. Differential expression of wheat genes was undertaken, comparing Pt‐ to mock‐inoculated and Pt‐ to Pt‐inoculated time points. Analysis of gene expression indicated a strong response to Pt, which was fully active by 6 h after inoculation (hai). More genes were downregulated than upregulated, particularly at 6 and 12 hai. Gene Ontology enrichment analysis indicated a shutting down of RNA and protein synthesis and an early effect on photosynthesis, with disruption of the electron transfer chain. Analyses of upregulated genes identified genes involved in ATP‐binding and protein kinase activity at 6 hai, supporting a rapid metabolic response to Pt infection. A general upregulation of genes involved in transport and metabolism indicated the need to relocate protein and organic‐based resources. Alignment of differentially expressed genes with the genomic regions defining four leaf rust APR quantitative trait loci (QTLs) in Toropi identified candidate resistance genes, including a sugar transporter, a receptor kinase and a seven‐transmembrane MLO family protein. In addition, 60 Pt genes were identified, 11 being annotated as potential effector proteins. [ABSTRACT FROM AUTHOR]