Wheat stem rust caused by Puccinia graminis f. sp. tritici (Pgt) had been a devastating foliar disease worldwide during the 20th century. With the emergence of Ug99 races, which are virulent to most stem rust resistance genes deployed in wheat varieties and advanced lines, stem rust has once again become a disease threatening global wheat production. Sr52, derived from Dasypyrum villosum and mapped to the long arm of 6V#3, is one of the few effective genes against Ug99 races. In this study, the wheat–D. villosum Robertsonian translocation T6AS·6V#3L, the only stock carrying Sr52 released to experimental and breeding programs so far, was crossed with a CS ph1b mutant to induce recombinants with shortened 6V#3L chromosome segments locating Sr52. Six independent homozygous recombinants with different segment sizes and breakpoints were developed and characterized using in situ hybridization and molecular markers analyses. Stem rust resistance evaluation showed that only three terminal recombinants (1381, 1380, and 1392) containing 8%, 22%, and 30% of the distal segment of 6V#3L, respectively, were resistant to stem rust. Thus, the gene Sr52 was mapped into 6V#3L bin FL 0.92–1.00. In addition, three molecular markers in the Sr52-located interval of 6V#3L were confirmed to be diagnostic markers for selection of Sr52 introgressed into common wheat. The newly developed small segment translocation lines with Sr52 and the identified molecular markers closely linked to Sr52 will be valuable for wheat disease breeding.

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is one of many severe diseases that threaten bread wheat (Triticum aestivum L.) yield and quality worldwide. The discovery and deployment of powdery mildew resistance genes (Pm) can prevent this disease epidemic in wheat. In a previous study, we transferred the powdery mildew resistance gene Pm57 from Aegilops searsii into common wheat and cytogenetically mapped the gene in a chromosome region with the fraction length (FL) 0.75–0.87, which represents 12% segment of the long arm of chromosome 2S^s#1. In this study, we performed RNA-seq using RNA extracted from leaf samples of three infected and mock-infected wheat-Ae. searsii 2S^s#1 introgression lines at 0, 12, 24, and 48 h after inoculation with Bgt isolates. Then we designed 79 molecular markers based on transcriptome sequences and physically mapped them to Ae. searsii chromosome 2S^s#1- in seven intervals. We used these markers to identify 46 wheat-Ae. searsii 2S^s#1 recombinants induced by ph1b, a deletion mutant of pairing homologous (Ph) genes. After analyzing the 46 ph1b-induced 2S^s#1L recombinants in the region where Pm57 is located with different Bgt-responses, we physically mapped Pm57 gene on the long arm of 2S^s#1 in a 5.13 Mb genomic region, which was flanked by markers X67593 (773.72 Mb) and X62492 (778.85 Mb). By comparative synteny analysis of the corresponding region on chromosome 2B in Chinese Spring (T. aestivum L.) with other model species, we identified ten genes that are putative plant defense-related (R) genes which includes six coiled-coil nucleotide-binding site-leucine-rich repeat (CNL), three nucleotide-binding site-leucine-rich repeat (NL) and a leucine-rich receptor-like repeat (RLP) encoding proteins. This study will lay a foundation for cloning of Pm57, and benefit the understanding of interactions between resistance genes of wheat and powdery mildew pathogens.

Wall-associated kinases (WAKs), a group of receptor-like kinases, have been found to play important roles in defending against pathogens and in various developmental processes. However, the importance of this family in wheat remains largely unknown. Wheat powdery mildew is caused by Blumeria graminis f. sp. tritici (Bgt), which initiates infection on the cell surface and forms haustoria inside the cell; therefore, the defense to Bgt involves extracellular and subsequently intracellular signals. In this study, WAKs were identified genome-wide and analyzed phylogenetically, and then a transmembrane WAK gene that putatively participated in pathogen-associated molecular pattern–triggered immunity and effector-triggered immunity to Bgt was functionally and evolutionarily investigated. In total, 1,193 WAKs were identified from wheat and its Gramineae relatives. Phylogenetic analysis indicated that WAKs expanded through tandem duplication or segment duplication. TaWAK7, from chromosome 2A, was identified as a Bgt-inducible gene both in susceptible and resistant materials, but it showed distinct responsive patterns. Functional analysis showed that TaWAK7 was involved in both the basal and resistance gene–mediated resistances. The specific gene structures and protein characteristics of TaWAK7, along with its orthologs, were characterized both in subgenomes of Triticum spp. and in the A genome of multiple wheat accessions, which revealed that TaWAK7 orthologs underwent complex evolution with frequent gene fusion and domain deletion. In addition, three cytoplasmic proteins interacting with TaWAK7 were indicated by yeast two-hybrid and bimolecular fluorescence complementation assays. Binding of TaWAK7 with these proteins could change its subcellular localization from the plasma membrane to the cytoplasm. This study provides a better understanding of the evolution of WAKs at the genomic level and TaWAK7 at the gene level and provides useful clues for further investigation of how WAKs transmit the extracellular signals to the cytoplasm to activate defense responses.

Powdery mildew of wheat, caused by Blumeria graminis f. sp. tritici, is a destructive disease of common wheat. Cultivation of resistant varieties is the most cost-effective disease management strategy. Previous studies reported that chromosome 3S^l#2 present in Chinese Spring (CS)–Aegilops longissima 3S^l#2(3B) disomic substitution line TA3575 conferred resistance to powdery mildew. In this study, we further located the powdery mildew resistance gene(s) to the short arm of chromosome 3S^l#2 (3S^l#2S) by evaluating for B. graminis f. sp. tritici resistance of newly developed CS–Ae. longissima 3S^l#2 translocation lines. Meanwhile, TA7545, a previously designated CS–Ae. longissima 3S^l#3 disomic addition line, was reidentified as an isochromosome 3S^l#3S addition line and evaluated to confer resistance to powdery mildew, thus locating the resistance gene(s) to the short arm of chromosome 3S^l#3 (3S^l#3S). Based on transcriptome sequences of TA3575, 10 novel chromosome 3S^lS-specific markers were developed, of which 5 could be used to distinguish between 3S^l#2S and 3S^l#3S derived from Ae. longissima accessions TL20 and TA1910 (TAM4) and the remaining 5 could identify both 3S^l#2S and 3S^l#3S. Also, CL897, one of five markers specific to both 3S^l#2S and 3S^l#3S, could be used to detect Pm13 located at chromosome 3S^l#1S from Ae. longissima accession TL01 in diverse wheat genetic backgrounds. The powdery mildew resistance genes on chromosomes 3S^l#2S and 3S^l#3S, the CS–Ae. longissima 3S^l#2 translocation lines, and the 3S^lS-specific markers developed in this study will facilitate the transfer of B. graminis f. sp. tritici resistance genes into common wheat and provide new germplasm resources for powdery mildew resistance breeding.