Wheat Stem Rust Resistance Gene Research Articles

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.

Single amino acid change alters specificity of the multi-allelic wheat stem rust resistance locus SR9

Most rust resistance genes thus far isolated from wheat have a very limited number of functional alleles. Here, we report the isolation of most of the alleles at wheat stem rust resistance gene locus SR9. The seven previously reported resistance alleles (Sr9a, Sr9b, Sr9d, Sr9e, Sr9f, Sr9g, and Sr9h) are characterised using a synergistic strategy. Loss-of-function mutants and/or transgenic complementation are used to confirm Sr9b, two haplotypes of Sr9e (Sr9e_h1 and Sr9e_h2), Sr9g, and Sr9h. Each allele encodes a highly related nucleotide-binding site leucine-rich repeat (NB-LRR) type immune receptor, containing an unusual long LRR domain, that confers resistance to a unique spectrum of isolates of the wheat stem rust pathogen. The only SR9 protein effective against stem rust pathogen race TTKSK (Ug99), SR9H, differs from SR9B by a single amino acid. SR9B and SR9G resistance proteins are also distinguished by only a single amino acid. The SR9 allelic series found in the B subgenome are orthologs of wheat stem rust resistance gene Sr21 located in the A subgenome with around 85% identity in protein sequences. Together, our results show that functional diversification of allelic variants at the SR9 locus involves single and multiple amino acid changes that recognize isolates of wheat stem rust.

High-resolution genetic mapping and identification of candidate genes for the wheat stem rust resistance gene Sr8155B1

Stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), threatens global wheat production. Development of cultivars with increased resistance to stem rust by identification, mapping, and deployment of resistance genes is the best strategy for controlling the disease. In this study, we performed fine mapping and characterization of the all-stage stem rust resistance (Sr) gene Sr8155B1 from the durum wheat line 8155-B1. In seedling tests of biparental populations, Sr8155B1 was effective against six Chinese Pgt races tested. In a segregating population of 5060 gametes, Sr8155B1 was mapped to a 0.06-cM region flanked by markers Pku2772 and Pku43365, corresponding to 1.5- and 2.7-Mb regions in the Svevo and Chinese Spring reference genomes. Both regions include several typical nucleotide-binding leucine-rich repeat (NLR) and protein kinase genes that represent candidate genes. Among them, three NLR genes and three receptor-like protein kinases were highly polymorphic between the parental lines and their transcripts were upregulated in the homozygous resistant line TdR2 relative to its susceptible sister line TdS4. Four markers (Pku2772, Pku43365, Pku2950, and Pku3721) developed in this study, together with seedling resistance responses, correctly predicted Sr8155B1 absence or presence in 78 tetraploid wheat genotypes tested. The presence of Sr8155B1 in tetraploid wheat accessions CItr 14916, PI 197492, and PI 197493 was confirmed by mapping in three F2 populations. The genetic map and linked markers developed in this study may accelerate the deployment of Sr8155B1-mediated resistance in wheat breeding programs.

The wheat stem rust resistance gene Sr43 encodes an unusual protein kinase

To safeguard bread wheat against pests and diseases, breeders have introduced over 200 resistance genes into its genome, thus nearly doubling the number of designated resistance genes in the wheat gene pool1. Isolating these genes facilitates their fast-tracking in breeding programs and incorporation into polygene stacks for more durable resistance. We cloned the stem rust resistance gene Sr43, which was crossed into bread wheat from the wild grass Thinopyrum elongatum2,3. Sr43 encodes an active protein kinase fused to two domains of unknown function. The gene, which is unique to the Triticeae, appears to have arisen through a gene fusion event 6.7 to 11.6 million years ago. Transgenic expression of Sr43 in wheat conferred high levels of resistance to a wide range of isolates of the pathogen causing stem rust, highlighting the potential value of Sr43 in resistance breeding and engineering.

Identification of wheat stem rust resistance genes in wheat cultivars from Hebei province, China.

Wheat stem rust is caused by Puccinia graminis f. sp. tritici. This major disease has been effectively controlled via resistance genes since the 1970s. The appearance and spread of new races of P. graminis f. sp. tritici (eg., Ug99, TKTTF, and TTRTF) have renewed the interest in identifying the resistance gene and breeding cultivars resistant to wheat stem rust. In this study, gene postulation, pedigree analysis, and molecular detection were used to determine the presence of stem rust resistance genes in 65 commercial wheat cultivars from Hebei Province. In addition, two predominant races 21C3CTHTM and 34MRGQM were used to evaluate the resistance of these cultivars at the adult-plant stage in 2021-2022. The results revealed that 6 Sr genes (namely, Sr5, Sr17, Sr24, Sr31, Sr32, Sr38, and SrTmp), either singly or in combination, were identified in 46 wheat cultivars. Overall, 37 wheat cultivars contained Sr31. Sr5 and Sr17 were present in 3 and 3 cultivars, respectively. Gao 5218 strong gluten, Jie 13-Ji 7369, and Kenong 1006 contained Sr24, Sr32, and Sr38, respectively. No wheat cultivar contained Sr25 and Sr26. In total, 50 (76.9%) wheat cultivars were resistant to all tested races of P. graminis f. sp. tritici in field test in 2021-2022. This study is important for breeding wheat cultivars with resistance to stem rust.

Identification of new sources of wheat stem rust resistance genes

Stem rust disease is widespread in the wheat-growing regions of Kazakhstan. Despite a large number of studies, the protection of wheat from the pathogen Puccinia graminis f. sp. tritici is considered one of the crucial problems. Chemical control has almost no impact on this disease and no exact result. The only effective way to control this disease is to cultivate resistant varieties and lines. Currently, 60 Sr resistance genes are known. Among them, Sr2, Sr22, Sr25/Lr19, Sr28, Sr36, and Sr39 gene sources are efficient at stem rust disease in different conditions of Kazakhstan. The molecular markers Xgwm533, CFA2019, PSY-E1, wPt-7004, Xgwm319 and Sr39#50 linked to Sr2, Sr22, Sr25/Lr19, Sr28, Sr36 and Sr39 were used, respectively. As a result of PCR analysis, the Sr2 gene was identified in six lines out of 16 lines, namely, GA951395-10-7/WX98D011-U38, Select, GA961565-27-6/KS99U673, GA961662-1-7/TAM107, VA01W-283/WX030513 and Sonalika. Five wheat lines were found as carriers of the Sr22 gene: Line c-19SB, Lutescens 7-04-4, Lutescens 220-03-45, GA961662-1-7/TAM107 and Line D 25 77. There are three lines that are carriers of Sr25/Lr19 gene sources, namely, Lutescens 220-03-45, Advance, and Line D 25 77. The Sr28 gene was identified in four wheat genotypes (GA951395-10-7/WX98D011-U38, Select, Advance and VA01W-283/WX03ASHTS0513) and the positive control W2691Sr2Bkt. An expected fragment (170 bp) for Xgwm319 properly for Sr36 gene was identified in four lines (GA951395-10-7/WX98D011-U38, Advance, VA01W-283/WX03ASHTS0513, GA961662-1-7/TAM107). The 10 wheat varieties were identified using primers Sr39 # 50R/F (Line C-19SB, Omskaya 37, Lutescens 7-04-4, Lutescens 220-03-45, Select, GA951395-10-7 / WX98D011-U38, Advance, GA961662-1-7 / TAM107’, VA01W-283/WX03ASHTS0513 and Line D 25 77). The studied sources of resistance can be used in breeding programs to create varieties of common wheat with durable resistance to stem rust.

The wheat stem rust resistance gene Sr43 encodes an unusual protein kinase

The wheat stem rust resistance gene Sr43 encodes an unusual protein kinase

Identification and characterization of Sr22b, a new allele of the wheat stem rust resistance gene Sr22 effective against the Ug99 race group.

SummaryWheat stem (or black) rust, caused by Puccinia graminis f. sp. tritici (Pgt), has been historically among the most devastating global fungal diseases of wheat. The recent occurrence and spread of new virulent races such as Ug99 have prompted global efforts to identify and isolate more effective stem rust resistance (Sr) genes. Here, we report the map‐based cloning of the Ug99‐effective SrTm5 gene from diploid wheat Triticum monococcum accession PI 306540 that encodes a typical coiled‐coil nucleotide‐binding leucine‐rich repeat protein. This gene, designated as Sr22b, is a new allele of Sr22 with a rare insertion of a large (13.8‐kb) retrotransposon into its second intron. Biolistic transformation of an ~112‐kb circular bacterial artificial chromosome plasmid carrying Sr22b into the susceptible wheat variety Fielder was sufficient to confer resistance to stem rust. In a survey of 168 wheat genotypes, Sr22b was present only in cultivated T. monococcum subsp. monococcum accessions but absent in all tested tetraploid and hexaploid wheat lines. We developed a diagnostic molecular marker for Sr22b and successfully introgressed a T. monococcum chromosome segment containing this gene into hexaploid wheat to accelerate its deployment and pyramiding with other Sr genes in wheat breeding programmes. Sr22b can be a valuable component of gene pyramids or transgenic cassettes combining different resistance genes to control this devastating disease.

Mapping and Characterization of a Wheat Stem Rust Resistance Gene in Durum Wheat "Kronos".

Wheat stem (or black) rust is one of the most devastating fungal diseases, threatening global wheat production. Identification, mapping, and deployment of effective resistance genes are critical to addressing this challenge. In this study, we mapped and characterized one stem rust resistance (Sr) gene from the tetraploid durum wheat variety Kronos (temporary designation SrKN). This gene was mapped on the long arm of chromosome 2B and confers resistance to multiple virulent Pgt races, such as TRTTF and BCCBC. Using a large mapping population (3,366 gametes), we mapped SrKN within a 0.29 cM region flanked by the sequenced-based markers pku4856F2R2 and pku4917F3R3, which corresponds to 5.6- and 7.2-Mb regions in the Svevo and Chinese Spring reference genomes, respectively. Both regions include a cluster of nucleotide binding leucine-repeat (NLR) genes that likely includes the candidate gene. An allelism test failed to detect recombination between SrKN and the previously mapped Sr9e gene. This result, together with the similar seedling resistance responses and resistance profiles, suggested that SrKN and Sr9e may represent the same gene. We introgressed SrKN into common wheat and developed completely linked markers to accelerate its deployment in the wheat breeding programs. SrKN can be a valuable component of transgenic cassettes or gene pyramids that includes multiple resistance genes to control this devastating disease.

Extensive Genetic Variation at the Sr22 Wheat Stem Rust Resistance Gene Locus in the Grasses Revealed Through Evolutionary Genomics and Functional Analyses.

In the last 20 years, severe wheat stem rust outbreaks have been recorded in Africa, Europe, and Central Asia. This previously well controlled disease, caused by the fungus Puccinia graminis f. sp. tritici, has reemerged as a major threat to wheat cultivation. The stem rust (Sr) resistance gene Sr22 encodes a nucleotide-binding and leucine-rich repeat receptor which confers resistance to the highly virulent African stem rust isolate Ug99. Here, we show that the Sr22 gene is conserved among grasses in the Triticeae and Poeae lineages. Triticeae species contain syntenic loci with single-copy orthologs of Sr22 on chromosome 7, except Hordeum vulgare, which has experienced major expansions and rearrangements at the locus. We also describe 14 Sr22 sequence variants obtained from both Triticum boeoticum and the domesticated form of this species, T. monococcum, which have been postulated to encode both functional and nonfunctional Sr22 alleles. The nucleotide sequence analysis of these alleles identified historical sequence exchange resulting from recombination or gene conversion, including breakpoints within codons, which expanded the coding potential at these positions by introduction of nonsynonymous substitutions. Three Sr22 alleles were transformed into wheat cultivar Fielder and two postulated resistant alleles from Schomburgk (hexaploid wheat introgressed with T. boeoticum segment carrying Sr22) and T. monococcum accession PI190945, respectively, conferred resistance to P. graminis f. sp. tritici race TTKSK, thereby unequivocally confirming Sr22 effectiveness against Ug99. The third allele from accession PI573523, previously believed to confer susceptibility, was confirmed as nonfunctional against Australian P. graminis f. sp. tritici race 98-1,2,3,5,6.[Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Correction to: Mapping and characterization of wheat stem rust resistance genes SrTm5 and Sr60 from Triticum monococcum.

In the original publication, the IWGSC assembly is incorrectly referenced.

Temperature‐sensitive wheat stem rust resistance gene Sr15 is effective against Puccinia graminis f. sp. tritici race TTKSK

The wheat stem rust fungus, Puccinia graminis f. sp. tritici (Pgt), race TTKSK and related races pose a serious threat to world wheat production. Knowing the effectiveness of wheat stem rust resistance (Sr) genes against Pgt race TTKSK is fundamental in mitigating this threat through resistance breeding. Sr15 was previously identified as being ineffective against Pgt race TTKSK. Here, multirace disease phenotyping data, linkage analyses, allelism testing and haplotype analyses are presented to support the conclusion that Sr15 is effective against Pgt race TTKSK. Resistance to race TTKSK was mapped to Sr15 in a biparental population. Thirty‐two accessions with Sr15 displayed seedling resistance phenotypes against race TTKSK. However, these accessions were susceptible as seedlings at high temperatures (22–25 °C), consistent with previous reports that the interaction between avirulent Pgt isolates and Sr15 is temperature‐sensitive. Markers STS638, wri4 and KASP_IWB30995 were found to predict the presence of Sr15, suggesting the utility of these assays for marker‐assisted selection in breeding programmes. The effectiveness of Sr15 to specific Pgt races and temperatures makes it a less‐desirable TTKSK‐effective gene. Wheat lines assayed as resistant to race TTKSK at the seedling stage may possess Sr15 and breeders should be aware of the limitations of Sr15 for conferring stem rust resistance.

Genetic Stability of Wheat Stem Rust Resistance Genes under Egyptian Field Conditions

Genetic Stability of Wheat Stem Rust Resistance Genes under Egyptian Field Conditions

Identification and characterization of wheat stem rust resistance gene Sr21 effective against the Ug99 race group at high temperature.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a devastating foliar disease. The Ug99 race group has combined virulence to most stem rust (Sr) resistance genes deployed in wheat and is a threat to global wheat production. Here we identified a coiled-coil, nucleotide-binding leucine-rich repeat protein (NLR) completely linked to the Ug99 resistance gene Sr21 from Triticum monococcum. Loss-of-function mutations and transgenic complementation confirmed that this gene is Sr21. Sr21 transcripts were significantly higher at high temperatures, and this was associated with significant upregulation of pathogenesis related (PR) genes and increased levels of resistance at those temperatures. Introgression of Sr21 into hexaploid wheat resulted in lower levels of resistance than in diploid wheat, but transgenic hexaploid wheat lines with high levels of Sr21 expression showed high levels of resistance. Sr21 can be a valuable component of transgenic cassettes or gene pyramids combining multiple resistance genes against Ug99.

Discovery of a Novel Stem Rust Resistance Allele in Durum Wheat that Exhibits Differential Reactions to Ug99 Isolates.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici Eriks. & E. Henn, can incur yield losses in susceptible cultivars of durum wheat, Triticum turgidum ssp. durum (Desf.) Husnot. Although several durum cultivars possess the stem rust resistance gene Sr13, additional genes in durum wheat effective against emerging virulent races have not been described. Durum line 8155-B1 confers resistance against the P. graminis f. sp. tritici race TTKST, the variant race of the Ug99 race group with additional virulence to wheat stem rust resistance gene Sr24. However, 8155-B1 does not confer resistance to the first-described race in the Ug99 race group: TTKSK. We mapped a single gene conferring resistance in 8155-B1 against race TTKST, Sr8155B1, to chromosome arm 6AS by utilizing Rusty/8155-B1 and Rusty*2/8155-B1 populations and the 90K Infinium iSelect Custom bead chip supplemented by KASP assays. One marker, KASP_6AS_IWB10558, cosegregated with Sr8155B1 in both populations and correctly predicted Sr8155B1 presence or absence in 11 durum cultivars tested. We confirmed the presence of Sr8155B1 in cultivar Mountrail by mapping in the population Choteau/Mountrail. The marker developed in this study could be used to predict the presence of resistance to race TTKST in uncharacterized durum breeding lines, and also to combine Sr8155B1 with resistance genes effective to Ug99 such as Sr13. The map location of Sr8155B1 cannot rule out the possibility that this gene is an allele at the Sr8 locus. However, race specificity indicates that Sr8155B1 is different from the known alleles Sr8a and Sr8b.

Discovery and characterization of two new stem rust resistance genes in Aegilops sharonensis

Key messageWe identified two novel wheat stem rust resistance genes, Sr-1644-1Sh and Sr-1644-5Sh in Aegilops sharonensis that are effective against widely virulent African races of the wheat stem rust pathogen.Stem rust is one of the most important diseases of wheat in the world. When single stem rust resistance (Sr) genes are deployed in wheat, they are often rapidly overcome by the pathogen. To this end, we initiated a search for novel sources of resistance in diverse wheat relatives and identified the wild goatgrass species Aegilops sharonesis (Sharon goatgrass) as a rich reservoir of resistance to wheat stem rust. The objectives of this study were to discover and map novel Sr genes in Ae. sharonensis and to explore the possibility of identifying new Sr genes by genome-wide association study (GWAS). We developed two biparental populations between resistant and susceptible accessions of Ae. sharonensis and performed QTL and linkage analysis. In an F6 recombinant inbred line and an F2 population, two genes were identified that mapped to the short arm of chromosome 1Ssh, designated as Sr-1644-1Sh, and the long arm of chromosome 5Ssh, designated as Sr-1644-5Sh. The gene Sr-1644-1Sh confers a high level of resistance to race TTKSK (a member of the Ug99 race group), while the gene Sr-1644-5Sh conditions strong resistance to TRTTF, another widely virulent race found in Yemen. Additionally, GWAS was conducted on 125 diverse Ae. sharonensis accessions for stem rust resistance. The gene Sr-1644-1Sh was detected by GWAS, while Sr-1644-5Sh was not detected, indicating that the effectiveness of GWAS might be affected by marker density, population structure, low allele frequency and other factors.

Markers Linked to Wheat Stem Rust Resistance Gene Sr11 Effective to Puccinia graminis f. sp. tritici Race TKTTF.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici, can cause severe yield losses on susceptible wheat varieties and cultivars. Although stem rust can be controlled by the use of genetic resistance, population dynamics of P. graminis f. sp. tritici can frequently lead to defeat of wheat stem rust resistance genes. P. graminis f. sp. tritici race TKTTF caused a severe epidemic in Ethiopia on Ug99-resistant 'Digalu' in 2013 and 2014. The gene Sr11 confers resistance to race TKTTF and is present in 'Gabo 56'. We identified seven single-nucleotide polymorphism (SNP) markers linked to Sr11 from a cross between Gabo 56 and 'Chinese Spring' exploiting a 90K Infinium iSelect Custom beadchip. Five SNP markers were validated on a 'Berkut'/'Scalavatis' population that segregated for Sr11, using KBioscience competitive allele-specific polymerase chain reaction (KASP) assays. Two of the SNP markers, KASP_6BL_IWB10724 and KASP_6BL_IWB72471, were predictive of Sr11 among wheat genetic stocks, cultivars, and breeding lines from North America, Ethiopia, and Pakistan. These markers can be utilized to select for Sr11 in wheat breeding and to detect the presence of Sr11 in uncharacterized germplasm.

Virulent Race Overcome Wheat Stem Rust Resistance Gene Sr27 in Egypt

Triticale is an excellent source of resistance to wheat stem rust, caused by Puccinia graminis f.sp. tritici. Stem rust historically is one of the most destructive diseases of wheat (Triticum aestivum L.) worldwide. Deployment of resistant genotypes successfully prevented rust epidemics over the past several decades. This is primarily due to the on-going incorporation of effective stem rust resistance genes into new wheat breeding materials; this may place the pathogen under great selection pressure. Although Sr27 has little effect on the commercial wheat production, it has a major impact on the commercial triticale cultivars. In Egypt, high levels of stem rust infection were observed on wheat cultivar (Cooring) which carrying Sr27 grown at Gemmiza Research Station during May 2015. Samples collected from rusted stems of wheat cultivar (Cooring) were cut and artificially inoculated on primary leaves of 8-day-old seedlings of highly susceptible wheat Cv. Morocco. Four single pustules were collected and race analysis was done. Results showed that out of the four identified races, race QQQCM was the most virulent one wheat seedling carrying Sr27 with infection type (4), whereas the rest of races were a virulent to Sr27. This race (QQQCM) first time toappear in Egypt from previous identified races. Regarding to the performance of 20 stem rust resistant genes at seedling stage, (Sr 9e,7b,8a,9g,30,17,9a,9d,10,31and Sr38) were resistant to race QQQCM, meanwhile the rest Srs were susceptible

The wheat Sr50 gene reveals rich diversity at a cereal disease resistance locus.

We identify the wheat stem rust resistance gene Sr50 (using physical mapping, mutation and complementation) as homologous to barley Mla, encoding a coiled-coil nucleotide-binding leucine-rich repeat (CC-NB-LRR) protein. We show that Sr50 confers a unique resistance specificity different from Sr31 and other genes on rye chromosome 1RS, and is effective against the broadly virulent Ug99 race lineage. Extensive haplotype diversity at the rye Sr50 locus holds promise for mining effective resistance genes.

Fine mapping and characterization of Sr21, a temperature-sensitive diploid wheat resistance gene effective against the Puccinia graminis f. sp. tritici Ug99 race group.

The diploid wheat stem rust resistance gene Sr21 confers temperature-sensitive resistance to isolates of the Ug99 group and maps to the middle of the long arm of chromosome 2A (m). A race of Puccinia graminis f. sp. tritici, the causal pathogen of stem rust of wheat, known as Ug99, and its variants, are virulent to plants carrying stem rust resistance genes currently deployed in most wheat cultivars worldwide. Therefore, identification, mapping and deployment of effective resistance genes are critical to reduce this threat. Resistance gene Sr21 identified in diploid wheat T. monococcum can be effective against races from the Ug99 race group, but both susceptible and partial resistant reactions have been reported in previous studies. To clarify this conflicting information we screened four monogenic lines with Sr21 and four susceptible controls with 16 Pgt isolates including five isolates of the Ug99 race group under three different temperatures and three different photoperiods. We observed that, temperature influences the interaction between monogenic lines with Sr21 and Ug99 race group isolates, and may be one source of previous inconsistencies. This result indicates that, although Sr21 confers partial resistance against Ug99, its effectiveness can be modulated by environmental conditions and should not be deployed alone. Using two large diploid wheat-mapping populations (total 3,788 F2 plants) we mapped Sr21 approximately 50 cM from the centromere on the long arm of chromosome 2A(m) within a 0.20 cM interval flanked by sequence-based markers FD527726 and EX594406. The closely linked markers identified in this study will be useful to reduce the T. monococcum segments introgressed into common wheat, accelerate Sr21 deployment in wheat breeding programs, and facilitate the map-based cloning of this gene.