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Summary: Powdery mildew poses severe threats to wheat production. The most sustainable way to control this disease is through planting resistant cultivars.We report the map‐based cloning of the powdery mildew resistance allele Pm5e from a Chinese wheat landrace. We applied a two‐step bulked segregant RNA sequencing (BSR‐Seq) approach in developing tightly linked or co‐segregating markers to Pm5e. The first BSR‐Seq used phenotypically contrasting bulks of recombinant inbred lines (RILs) to identify Pm5e‐linked markers. The second BSR‐Seq utilized bulks of genetic recombinants screened from a fine‐mapping population to precisely quantify the associated genomic variation in the mapping interval, and identified the Pm5e candidate genes.The function of Pm5e was validated by transgenic assay, loss‐of‐function mutants and haplotype association analysis. Pm5e encodes a nucleotide‐binding domain leucine‐rich‐repeat‐containing (NLR) protein. A rare nonsynonymous single nucleotide variant (SNV) within the C‐terminal leucine rich repeat (LRR) domain is responsible for the gain of powdery mildew resistance function of Pm5e, an allele endemic to wheat landraces of Shaanxi province of China.Results from this study demonstrate the value of landraces in discovering useful genes for modern wheat breeding. The key SNV associated with powdery mildew resistance will be useful for marker‐assisted selection of Pm5e in wheat breeding programs. [ABSTRACT FROM AUTHOR]

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most destructive diseases that pose a great threat to wheat production. Wheat landraces represent a rich source of powdery mildew resistance. Here, we report the map-based cloning of powdery mildew resistance gene Pm24 from Chinese wheat landrace Hulutou. It encodes a tandem kinase protein (TKP) with putative kinase-pseudokinase domains, designated WHEAT TANDEM KINASE 3 (WTK3). The resistance function of Pm24 was validated by transgenic assay, independent mutants, and allelic association analyses. Haplotype analysis revealed that a rare 6-bp natural deletion of lysine-glycine codons, endemic to wheat landraces of Shaanxi Province, China, in the kinase I domain (Kin I) of WTK3 is critical for the resistance function. Transgenic assay of WTK3 chimeric variants revealed that only the specific two amino acid deletion, rather than any of the single or more amino acid deletions, in the Kin I of WTK3 is responsible for gaining the resistance function of WTK3 against the Bgt fungus. Powdery mildew is a major threat to world wheat yields. Here the authors describe the map-based cloning of Pm24, a gain-of-function powdery mildew resistance allele that encodes a tandem kinase-pseudokinase protein with a deletion in a kinase domain that is endemic to certain wheat landraces. [ABSTRACT FROM AUTHOR]