Molecular detection of a gene effective against powdery mildew in the wheat cultivar Liangxing 66

Since it was commercialized in 2008, Liangxing 66 is one of the most widely grown cultivars of wheat (Triticum aestivum L.) in winter and facultative wheat-producing regions in northern China. This cultivar displays broad-spectrum resistance to isolates of powdery mildew. To identify the powdery mildew resistance gene in Liangxing 66, genetic analysis and molecular mapping were conducted using the F₂ populations and F_2:3 families derived from the reciprocal crosses of Liangxing 66 and the susceptible cultivar Jingshuang 16. A single dominant gene, tentatively designated PmLX66, conferred resistance in Liangxing 66 to the powdery mildew isolate E09. The results of molecular mapping indicated that this gene was located on the short arm of chromosome 5D and flanked by SCAR203 and Xcfd81 at genetic distances of 0.4 and 2.8?cM, respectively, which is similar to the position of locus Pm2. However, PmLX66 and Pm2 showed different reactions to five of the 42 isolates of powdery mildew tested. Together, these results indicated that PmLX66 was most likely an allele of Pm2. Based on its superior yield and agronomic performance, in combination with powdery mildew resistance, Liangxing 66 is useful as a promising parent for control of powdery mildew and for the development of new disease-resistant cultivars.     

The Podosphaera fusca TUB2 gene,a molecular “Swiss Army knife” with multiple applications in powdery mildew research

The powdery mildew fungus Podosphaera fusca (synonym Podosphaera xanthii) is the main causal agent of cucurbit powdery mildew and one of the most important limiting factors for cucurbit production worldwide. Despite the fungus' economic importance, very little is known about the physiological and molecular processes involved in P. fusca biology and pathogenesis. In this study, we isolated and characterised the β-tubulin-encoding gene of P. fusca (PfTUB2) to develop molecular tools with different applications in powdery mildew research. PfTUB2 is predicted to encode a protein of 447 amino acid residues. The coding region is interrupted by six introns that occur at approximately the same positions as the introns present in other fungal TUB2-like genes. Once cloned, the PfTUB2 sequence information was used in different applications. Our results showed that the TUB2 gene is a good marker for molecular phylogenetics in powdery mildew fungi but it is unsuitable for the analysis of intraspecific diversity in P. fusca. The expression of PfTUB2 was proven to be stable in different temperature conditions, supporting its use as a reference gene in quantitative gene expression studies. Furthermore, an allele-specific PCR assay for the detection of resistance to methyl-2-benzimidazole carbamate (MBC) fungicides in P. fusca was developed based on the correlation between the single amino acid change E198A in β-tubulin and the MBC resistance phenotype. Lastly, PfTUB2 was used as a target gene in the development of a high-throughput method to quantify fungal growth in plant tissues.     

Molecular mapping of a recessive powdery mildew resistance gene in spelt wheat cultivar Hubel

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most important wheat diseases worldwide. The basis for wheat powdery mildew resistance breeding consists of screening diversified host genetic resources with a range of races of the powdery mildew pathogen. Spelt wheat (Triticum aestivum ssp. spelta 2n = 6x = 42, AABBDD) is a close relative of common wheat (T. aestivum ssp. aestivum) and contains several known disease resistance genes, including Pm1d, Yr5, and Lr65. Here, we report the identification and mapping of a powdery mildew resistance gene in spelt wheat cultivar Hubel, which was introduced to China from Europe and is resistant to Chinese Bgt isolate E09 at the seedling stage. Genetic analysis of a recombinant inbred line population derived from a cross of Hubel and a susceptible early maturing mutant line indicated that Hubel possessed a recessive powdery mildew resistance gene (temporarily designated MlHubel). Markers linked to MlHubel were identified using bulked segregant analysis, simple sequence repeat, and expressed sequence tag-derived sequence tagged site methods. The linked markers were physically located on wheat chromosome 2D. Comparative genomic analysis indicated that the genetic interval covering MlHubel in wheat is highly colinear with the corresponding regions on Brachypodium distachyon chromosome 5 and Oryza sativa chromosome 4. Accordingly, the genetic map of MlHubel was established in comparison with B. distachyon 5L and O. sativa 4L, with the closest marker Xgwm265 being 0.4 cM from MlHubel. The identification of the recessive powdery mildew gene in spelt wheat suggests the potential of this accession along with its closely linked markers in breeding for resistance to powdery mildew.     

The antagonistic strain Bacillus subtilis UMAF6639 also confers protection to melon plants against cucurbit powdery mildew by activation of jasmonate-and salicylic acid-dependent defence responses

Biological control of plant diseases has gained acceptance in recent years. Bacillus subtilis UMAF6639 is an antagonistic strain specifically selected for the efficient control of the cucurbit powdery mildew fungus Podosphaera fusca, which is a major threat to cucurbits worldwide. The antagonistic activity relies on the production of the antifungal compounds iturin and fengycin. In a previous study, we found that UMAF6639 was able to induce systemic resistance (ISR) in melon and provide additional protection against powdery mildew. In the present work, we further investigated in detail this second mechanism of biocontrol by UMAF6639. First, we examined the signalling pathways elicited by UMAF6639 in melon plants, as well as the defence mechanisms activated in response to P. fusca. Second, we analysed the role of the lipopeptides produced by UMAF6639 as potential determinants for ISR activation. Our results demonstrated that UMAF6639 confers protection against cucurbit powdery mildew by activation of jasmonate- and salicylic acid-dependent defence responses, which include the production of reactive oxygen species and cell wall reinforcement. We also showed that surfactin lipopeptide is a major determinant for stimulation of the immune response. These results reinforce the biotechnological potential of UMAF6639 as a biological control agent.     

Isolation and evaluation of antagonistic bacteria towards the cucurbit powdery mildew fungus<Emphasis Type="Italic"> Podosphaera fusca</Emphasis>

Powdery mildew is one of the most important limiting factors for cucurbits production in Spain, its management being strongly dependent on chemicals. The aim of this work was to evaluate the possibility of exploiting antagonistic bacteria in the biological control of the cucurbit powdery mildew fungus Podosphaera fusca (syn. Sphaerotheca fusca). Among a collection of bacterial strains isolated from distinct cucurbit powdery mildew diseased plants and rhizospheric soils, four isolates were selected, by means of a screening method based on antibiotic production, and identified as Bacillus spp. These isolates proved to be efficacious in the control of cucurbit powdery mildew in in vitro detached leaves and seedling biocontrol assays, where reductions of disease severity of up to 80% were obtained. Furthermore, bacterial populations on melon leaves remained at similar levels (10⁵ cfu cm^–2) over the 16-day period studied and, as observed by scanning electron microscopy analysis, they were able to establish microcolonies associated with an extracellular matrix, which reveals that these isolates efficiently colonize melon phylloplane. These results indicate that the bacterial isolates selected are promising candidates for biological control agents of cucurbit powdery mildew in southern Spain.     

Genetic linkage map of melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) and localization of a major QTL for powdery mildew resistance

Powdery mildew caused by Podosphaera xanthii has become a major problem in melon since it occurs all year round irrespective of the growing system. The TGR-1551 melon genotype was found to be resistant to several melon diseases, among them powdery mildew. However, the corresponding resistance genes have been never mapped. We constructed an integrated genetic linkage map using an F2 population derived from a cross between the multi-resistant genotype TGR-1551 and the susceptible Spanish cultivar ‘Bola de Oro’. The map spans 1,284.9 cM, with an average distance of 3.6 cM among markers, and consists of 354 loci (188 AFLP, 39 RAPD, 111 SSR, 14 SCAR/CAPS/dCAPS, and two phenotypic traits) distributed in 14 linkage groups. QTL analysis identified one major QTL (Pm-R) on LG V for resistance to races 1, 2, and 5 of powdery mildew. The PM4-CAPS marker is closely linked to the Pm-R QTL at a genetic distance of 1.9 cM, and the PM3-CAPS marker is located within the support interval of this QTL. These codominant markers, together with the map information reported here, could be used for melon breeding, and particularly for genotyping selection of resistance to powdery mildew in this vegetable crop species.     

Pm50: a new powdery mildew resistance gene in common wheat derived from cultivated emmer

Fungal diseases of wheat, including powdery mildew, cause significant crop, yield and quality losses throughout the world. Knowledge of the genetic basis of powdery mildew resistance will greatly support future efforts to develop and cultivate resistant cultivars. Studies were conducted on cultivated emmer-derived wheat line K2 to identify genes involved in powdery mildew resistance at the seedling and adult plant growth stages using a BC₁ doubled haploid population derived from a cross between K2 and susceptible cultivar Audace. A single gene was located distal to microsatellite marker Xgwm294 on the long arm of chromosome 2A. Quantitative trait loci (QTL) analysis indicated that the gene was also effective at the adult plant stage, explaining up to 79.0 % of the variation in the progeny. Comparison of genetic maps indicated that the resistance gene in K2 was different from Pm4, the only other formally named resistance gene located on chromosome 2AL, and PmHNK54, a gene derived from Chinese germplasm. The new gene was designated Pm50.     

Sensibilité du Melon àSphaerotheca fuliginea en Fonction de L'Etage Foliaire

Effect of leaf position on the susceptibility of melon plants to artificial infection with powdery mildew, Sphaerotheca fuliginea The leaf position of melon plants seems to play a role on their susceptibility when they are artificially infected with powdery mildew Sphaerotheca fuliginea. The cotyledons are generally very susceptible, while the first leaf relatively resistant; the susceptibility again continues up to the 4th–5th leaf (but less susceptible than the cotyledons) and then after it decreases; these results can be obtained on plants in greenhouse or on detached leaves in Petri dishes. From this observation, we think that the screening of melon genotypes for resistance to powdery mildew can be evaluated neither on the cotyledons which are very susceptible nor on the first leaf which is resistant, but on the third leaf which is moderately susceptible. In fact, there is a good correlation between the reaction of the third leaf and the resistance or susceptibility of genotypes.     

Identification of QTLs for resistance to powdery mildew and SSR markers diagnostic for powdery mildew resistance genes in melon (Cucumis melo L.)

Powdery mildew caused by Podosphaera xanthii is an important foliar disease in melon. To find molecular markers for marker-assisted selection, we constructed a genetic linkage map of melon based on a population of 93 recombinant inbred lines derived from crosses between highly resistant AR 5 and susceptible ‘Earl’s Favourite (Harukei 3)’. The map spans 877 cM and consists of 167 markers, comprising 157 simple sequence repeats (SSRs), 7 sequence characterized amplified region/cleavage amplified polymorphic sequence markers and 3 phenotypic markers segregating into 20 linkage groups. Among them, 37 SSRs and 6 other markers were common to previous maps. Quantitative trait locus (QTL) analysis identified two loci for resistance to powdery mildew. The effects of these QTLs varied depending on strain and plant stage. The percentage of phenotypic variance explained for resistance to the pxA strain was similar between QTLs (R ² = 22–28%). For resistance to pxB strain, the QTL on linkage group (LG) XII was responsible for much more of the variance (41–46%) than that on LG IIA (12–13%). The QTL on LG IIA was located between two SSR markers. Using an independent population, we demonstrated the effectiveness of these markers. This is the first report of universal and effective markers linked to a gene for powdery mildew resistance in melon.     

A mutation in a coproporphyrinogen III oxidase gene confers growth inhibition, enhanced powdery mildew resistance and powdery mildew-induced cell death in Arabidopsis

Key message

A gene encoding a coproporphyrinogen III oxidase mediates disease resistance in plants by the salicylic acid pathway.

Abstract

A number of genes that regulate powdery mildew resistance have been identified in Arabidopsis, such as ENHANCED DISEASE RESISTANCE 1 to 3 (EDR1 to 3). To further study the molecular interactions between the powdery mildew pathogen and Arabidopsis, we isolated and characterized a mutant that exhibited enhanced resistance to powdery mildew. The mutant also showed dramatic powdery mildew-induced cell death as well as growth defects and early senescence in the absence of pathogens. We identified the affected gene by map-based cloning and found that the gene encodes a coproporphyrinogen III oxidase, a key enzyme in the tetrapyrrole biosynthesis pathway, previously known as LESION INITIATION 2 (LIN2). Therefore, we designated the mutant lin2-2. Further studies revealed that the lin2-2 mutant also displayed enhanced resistance to Hyaloperonospora arabidopsidis (H.a.) Noco2. Genetic analysis showed that the lin2-2-mediated disease resistance and spontaneous cell death were dependent on PHYTOALEXIN DEFICIENT 4 (PAD4), SALICYLIC ACID INDUCTION-DEFICIENT 2 (SID2), and NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1), which are all involved in salicylic acid signaling. Furthermore, the relative expression levels of defense-related genes were induced after powdery mildew infection in the lin2-2 mutant. These data indicated that LIN2 plays an important role in cell death control and defense responses in plants.     

Isolation and selection of plant growth-promoting rhizobacteria as inducers of systemic resistance in melon

Backgroud and aims

Powdery mildew elicited by Podosphaera fusca is an important threat to cucurbits. In order to find alternatives to the current use of chemicals, we examined the potential use of plant growth-promoting rhizobacteria (PGPR) for controlling the disease by induction of systemic resistance in the host plant.

Methods

A collection of Bacillus and Pseudomonas strains from different origins was studied, including strains isolated from roots of disease-free melon plants obtained from a greenhouse plagued by powdery mildew. The selection of best candidates was based on the evaluation of different traits commonly associated with PGPR, such as antifungal and siderophore production, swimming and swarming motilities, biofilm formation, auxin production and promotion of root development.

Results

Three Bacillus strains, B. subtilis UMAF6614 and UMAF6639 and B. cereus UMAF8564, and two Pseudomonas fluorescens strains, UMAF6031 and UMAF6033, were selected after ranking the strains using a nonparametric statistics test. Applied to melon seedlings, the selected strains were able to promote plant growth, increasing fresh weight up to 30%. Furthermore, these strains provided protection against powdery mildew and also against angular leaf spot caused by Pseudomonas syringae pv. lachrymans, with disease reductions of up to 60%.

Conclusions

These results suggest that the use of ISR-promoting PGPR could be a promising strategy for the integrated control of cucurbit powdery mildew and other cucurbit diseases.     

Mapping of novel powdery mildew resistance gene(s) from <Emphasis Type="Italic">Agropyron cristatum</Emphasis> chromosome 2P

Key message

A physical map of Agropyron cristatum 2P chromosome was constructed for the first time and the novel powdery mildew resistance gene(s) from chromosome 2P was(were) also mapped.

Abstract

Agropyron cristatum (L.) Gaertn. (2n = 28, PPPP), a wild relative of common wheat, is highly resistant to powdery mildew. Previous studies showed that wheat-A. cristatum 2P disomic addition line II-9-3 displayed high resistance to powdery mildew, and the resistance was attributable to A. cristatum chromosome 2P. To utilize and physically map the powdery mildew resistance gene(s), 15 wheat-A. cristatum 2P translocation lines and three A. cristatum 2P deletion lines with different chromosomal segment sizes, obtained from II-9-3 using ⁶⁰Co-γ ray irradiation, were characterized using cytogenetic and molecular marker analysis. A. cristatum 2P chromosomal segments in the translocations were translocated to different wheat chromosomes, including 1A, 4A, 5A, 6A, 7A, 1B, 2B, 3B, 7B, 3D, 4D, and 6D. A physical map of the 2P chromosome was constructed with 82 STS markers, consisting of nine bins with 34 markers on 2PS and eight bins with 48 markers on 2PL. The BC₁F₂ populations of seven wheat-A. cristatum 2P translocation lines (2PT-3, 2PT-4, 2PT-5, 2PT-6, 2PT-8, 2PT-9, and 2PT-10) were developed by self-pollination, tested with powdery mildew and genotyped with 2P-specific STS markers. From these results, the gene(s) conferring powdery mildew resistance was(were) located on 2PL bin FL 0.66–0.86 and 19 2P-specific markers were identified in this bin. Moreover, two new powdery mildew-resistant translocation lines (2PT-4 and 2PT-5) with small 2PL chromosome segments were obtained. The newly developed wheat lines with powdery mildew resistance and the closely linked molecular markers will be valuable for wheat disease breeding in the future.

     

TaRPP13-3, a CC-NBS-LRR-like gene located on chr 7D,promotes disease resistance to wheat powdery mildew in Brock

Disease resistance (R) gene, RPP13, plays an important role in the resistance of plants to pathogen infections; its function in resistance of wheat to powdery mildew remains unknown. In this study, a RNA-Seq technique was used to monitor expression of genes in susceptible wheat ‘Jing411’ and resistant near-isogenic line ‘BJ-1’ in response to powdery mildew infection. Overall, 413 differential expression genes were observed and identified as involved in disease resistance. RPP13 homologous gene on wheat chromosome 7D was preliminarily identified using the wheat 660K SNP chip. RPP13 was highly expressed in ‘BJ-1’ and encodes 1,027 amino acids, including CC, NB and LRR domain, termed TaRPP13-3. After inoculation with powdery mildew, expression of TaRPP13-3 in resistant wheat changed with time, but average expression was higher when compared to susceptible variety, thus indicating that TaRPP13-3 is involved in resistance to powdery mildew. Virus-induced gene silencing (VIGS) was used to inhibit expression of TaRPP13-3 in resistant parent ‘Brock’. Results indicated that silencing of TaRPP13-3 led to decreased disease resistance in ‘Brock’. Overall results of this study indicate that TaRPP13-3 gene is involved in the defence response of wheat to powdery mildew and plays a positive role in wheat powdery mildew interactions.     

Chromosomal Location and Comparative Genomics Analysis of Powdery Mildew Resistance Gene Pm51 in a Putative Wheat-Thinopyrum ponticum Introgression Line

Powdery mildew (PM) is a very destructive disease of wheat (Triticum aestivum L.). Wheat-Thinopyrum ponticum introgression line CH7086 was shown to possess powdery mildew resistance possibly originating from Th. ponticum. Genomic in situ hybridization and molecular characterization of the alien introgression failed to identify alien chromatin. To study the genetics of resistance, CH7086 was crossed with susceptible genotypes. Segregation in F₂ populations and F_2:3 lines tested with Chinese Bgt race E09 under controlled conditions indicated that CH7086 carries a single dominant gene for powdery mildew resistance. Fourteen SSR and EST-PCR markers linked with the locus were identified. The genetic distances between the locus and the two flanking markers were 1.5 and 3.2 cM, respectively. Based on the locations of the markers by nullisomic-tetrasomic and deletion lines of ‘Chinese Spring’, the resistance gene was located in deletion bin 2BL-0.89-1.00. Conserved orthologous marker analysis indicated that the genomic region flanking the resistance gene has a high level of collinearity to that of rice chromosome 4 and Brachypodium chromosome 5. Both resistance specificities and tests of allelism suggested the resistance gene in CH7086 was different from previously reported powdery mildew resistance genes on 2BL, and the gene was provisionally designated PmCH86. Molecular analysis of PmCH86 compared with other genes for resistance to Bgt in the 2BL-0.89-1.00 region suggested that PmCH86 may be a new PM resistance gene, and it was therefore designated as Pm51. The closely linked flanking markers could be useful in exploiting this putative wheat-Thinopyrum translocation line for rapid transfer of Pm51 to wheat breeding programs.     

Genetic analysis of a novel broad-spectrum powdery mildew resistance gene from the wheat-<Emphasis Type="Italic">Agropyron cristatum</Emphasis> introgression line Pubing 74

Main conclusion

A novel broad-spectrum powdery mildew resistance gene PmPB74 was identified in wheat- Agropyron cristatum introgression line Pubing 74. Development of wheat cultivars with broad-spectrum, durable resistance to powdery mildew has been restricted by lack of superior genetic resources. In this study, a wheat-A. cristatum introgression line Pubing 74, originally selected from a wide cross between the common wheat cultivar Fukuhokomugi (Fukuho) and Agropyron cristatum (L.) Gaertn (2n = 4x = 28; genome PPPP), displayed resistance to powdery mildew at both the seedling and adult stages. The putative alien chromosomal fragment in Pubing 74 was below the detection limit of genomic in situ hybridization (GISH), but evidence for other non-GISH-detectable introgressions was provided by the presence of three STS markers specific to A. cristatum. Genetic analysis indicated that Pubing 74 carried a single dominant gene for powdery mildew resistance, temporarily designated PmPB74. Molecular mapping showed that PmPB74 was located on wheat chromosome arm 5DS, and flanked by markers Xcfd81 and HRM02 at genetic distances of 2.5 and 1.7 cM, respectively. Compared with other lines with powdery mildew resistance gene(s) on wheat chromosome arm 5DS, Pubing 74 was resistant to all 28 Blumeria graminis f. sp tritici (Bgt) isolates from different wheat-producing regions of northern China. Allelism tests indicated that PmPB74 was not allelic to PmPB3558 or Pm2. Our work showed that PmPB74 is a novel gene with broad resistance to powdery mildew, and hence will be helpful in broadening the genetic basis of powdery mildew resistance in wheat.

     

Transfer and mapping of a gene conferring later-growth-stage powdery mildew resistance in a tetraploid wheat accession

Wheat powdery mildew is a severe foliar disease and causes significant yield losses in epidemic years. Breeding and using resistant cultivars is the most widely employed strategy to curb this disease. To identify and transfer powdery mildew resistance genes in wild emmer wheat accession TA1410 into common wheat, a resistant F3 line derived from the cross of TA1410 × durum wheat line Zhongyin1320 was crossed with common wheat cultivar Yangmai158. The homozygous resistant BC5F2 lines derived from the backcross with Yangmai158 exhibited susceptibility at seedling stage and conferred increasing resistance when the plants were closer to heading stage. In two segregating BC5F3 families investigated at heading stage, the segregation of the resistance fit a 3:1 ratio, suggesting that a single dominant gene controls the resistance. This resistance gene, designated HSM1, was mapped to the 0.6-cM Xmag5825.1–Xgwm344 interval on chromosome 7AL and co-segregated with Xrga-C3 and Xrga-C6. A mapping position comparison with other powdery mildew resistance genes on this chromosome suggested that HSM1 belongs to the Pm1 resistance gene cluster. HSM1 is a useful candidate gene for resistance breeding, particularly in winter-wheat growing areas.     

Identification and mapping of <Emphasis Type="Italic">MLIW30</Emphasis>, a novel powdery mildew resistance gene derived from wild emmer wheat

Powdery mildew, caused by Blumeria graminis f.sp. tritici (Bgt), is a destructive foliar disease of common wheat in areas with cool or maritime climates. Wild emmer wheat, Triticum turgidum ssp. dicoccoides, the progenitor of both domesticated tetraploid durum wheat and hexaploid bread wheat, harbors abundant genetic diversity related to resistance to powdery mildew that can be utilized for wheat improvement. An F₂ segregating population was obtained from a cross between resistant bread wheat line 2L6 and susceptible cultivar Liaochun 10, after which genetic analysis of F₂ and F₂-derived F₃ families was performed by inoculating plants with isolate Bgt E09. The results of this experiment demonstrated that powdery mildew resistance in 2L6, which was derived from wild emmer wheat accession IW30, was controlled by a single dominant gene, temporarily designated MLIW30. Nineteen SSR markers and two STS markers linked with MLIW30 were acquired by applying bulked segregant analysis. Finally, MLIW30 was located to the long arm of chromosome 4A and found to be flanked by simple sequence repeat markers XB1g2000.2 and XB1g2020.2 at 0.1 cM. Because no powdery mildew resistance gene in or derived from wild emmer wheat has been reported in wheat chromosome 4A, MLIW30 might be a novel Pm gene.     

Genetic and comparative genomics mapping reveals that a powdery mildew resistance gene Ml3D232 originating from wild emmer co-segregates with an NBS-LRR analog in common wheat (Triticum aestivum L.)

Powdery mildew caused by Blumeria graminis f. sp. tritici is one of the most important wheat diseases worldwide and breeding for resistance using diversified disease resistance genes is the most promising approach to prevent outbreaks of powdery mildew. A powdery mildew resistance gene, originating from wild emmer wheat (Triticum turgidum var. dicoccoides) accessions collected from Israel, has been transferred into the hexaploid wheat line 3D232 through crossing and backcrossing. Inoculation results with 21 B. graminis f. sp. tritici races indicated that 3D232 is resistant to all of the powdery mildew isolates tested. Genetic analyses of 3D232 using an F₂ segregating population and F₃ families indicated that a single dominant gene, Ml3D232, confers resistance in the host seedling stage. By applying molecular markers and bulked segregant analysis (BSA), we have identified polymorphic simple sequence repeats (SSR), expressed sequence tags (EST) and derived sequence tagged site (STS) markers to determine that the Ml3D232 is located on chromosome 5BL bin 0.59–0.76. Comparative genetic analyses using mapped EST markers and genome sequences of rice and Brachypodium established co-linearity of the Ml3D232 genomic region with a 1.4 Mb genomic region on Brachypodium distachyon chromosome 4, and a 1.2 Mb contig located on the Oryza sativa chromosome 9. Our comparative approach enabled us to develop new EST–STS markers and to delimit the genomic region carrying Ml3D232 to a 0.8 cM segment that is collinear with a 558 kb region on B. distachyon. Eight EST markers, including an NBS-LRR analog, co-segregated with Ml3D232 to provide a target site for fine genetic mapping, chromosome landing and map-based cloning of the powdery mildew resistance gene. This newly developed common wheat germplasm provides broad-spectrum resistance to powdery mildew and a valuable resource for wheat breeding programs.