Pathotypes of Blumeria graminis f. sp. tritici,the causal agent of wheat powdery mildew from some regions of Iran

Wheat powdery mildew is controlled mainly by race-specific resistance. To be effective, breeding wheat for resistance to powdery mildew requires knowledge of virulence diversity in local populations of the pathogen. Isolates of Blumeria graminis, collected in 2009 and 2010 from three areas of Iranian production, were analysed for virulence using a host differential series comprised of 16 known genes conferring resistance to powdery mildew. The results showed that high-virulence frequencies to genes Pm1, Pm2, Pm4a, Pm5, Pm6, Pm7, Pm8 and Pm9 were found over both years and across all three areas. Virulence frequencies for Pm3a and Pm3b were intermediate, while virulence frequencies for Pm3a, Pm3c, Pm4a and Pm2, 6 were low. Genes Pm1, 2, 9 and Pm2, 4b, 8 were highly resistant in all regions. Virulence to Pm8 increased to high levels, while virulence to Pm4a decreased across the area surveyed from 2009 to 2010.     

Compatibility of Erysiphe graminis Hybrids f. sp. secalis X f. sp. Tritici with Wheat Lines Involving Resistance Genes from Rye

Compatibility of hybrid cultures Erysiphe graminis ff. sp. secalis (SI) ×tritici (t2) was tested in the laboratory with wheat cultivars involving different resistance genes and with two rye cultivars. Segregation was observed on wheat without resistance gene and with resistance genes Pm1, Pm3b and Pm3c compatible with t2, but not on wheat with resistance gene Pm2, Pm 3a, Pm 4a and Pm 5 incompatible with t2, nor on rye. It was obvious that S1 involves avirulence genes to Pm1, Pm2, Pm 3a, pm 3b, Pm 3c, Pm 4a, Pm 5. Segregation was found on wheat cultivars involving rye resistance genes Pm 7 (Transfed) and Pm 8 (Kavkaz), but cv. Transec (Pm7) was incompatible with all cultures used, because Transec involves another gene for resistance. The results indicate that hybridization between formae speciales secalis and tritici of the fungus can be a source of fungus compatibility with wheat with rye resistance, even in field conditions.     

Genetic characterization of powdery mildew resistance in U.S. hard winter wheat

Powdery mildew significantly affects grain yield and end-use quality of winter wheat in the southern Great Plains. Employing resistance resources in locally adapted cultivars is the most effective means to control powdery mildew. Two types of powdery mildew resistance exist in wheat cultivars, i.e., qualitative and quantitative. Qualitative resistance is controlled by major genes, is race-specific, is not durable, and is effective in seedlings and in adult plants. Quantitative resistance is controlled by minor genes, is non-race-specific, is durable, and is predominantly effective in adult plants. In this study, we found that the segregation of powdery mildew resistance in a population of recombinant inbred lines developed from a cross between the susceptible cultivar Jagger and the resistant cultivar 2174 was controlled by a major QTL on the short arm of chromosome 1A and modified by four minor QTLs on chromosomes 1B, 3B, 4A, and 6D. The major QTL was mapped to the genomic region where the Pm3 gene resides. Using specific PCR markers for seven Pm3 alleles, 2174 was found to carry the Pm3a allele. Pm3a explained 61% of the total phenotypic variation in disease reaction observed among seedlings inoculated in the greenhouse and adult plants grown in the field and subjected to natural disease pressure. The resistant Pm3a allele was present among 4 of 31 cultivars currently being produced in the southern Great Plains. The genetic effects of several minor loci varied with different developmental stages and environments. Molecular markers associated with these genetic loci would facilitate incorporating genetic resistance to powdery mildew into improved winter wheat cultivars.     

Chromosomal location of a gene suppressing powdery mildew resistance genes Pm8 and Pm17 in common wheat (Triticum aestivum L. em. Thell.)

The chromosomal location of a suppressor for the powdery mildew resistance genes Pm8 and Pm17 was determined by a monosomic set of the wheat cultivar Caribo. This cultivar carries a suppressor gene inhibiting the expression of Pm8 in cv Disponent and of Pm17 in line Helami-105. In disease resistance assessments, monosomic F₁ hybrids (2n=41) of Caribo x Disponent and Caribo x Helami-105 lacking chromosome 7D were resistant, whereas monosomic F₁ hybrids involving the other 20 chromosomes, as well as disomic F₁ hybrids (2n=42) of all cross combinations, were susceptible revealing that the suppressor gene for Pm8 and Pm17 is localized on chromosome 7D. It is suggested that genotypes without the suppressor gene be used for the exploitation of genes Pm8 and Pm17 in enhancing powdery mildew resistance in common wheat.     

Virulence analysis of wheat powdery mildew population (Erysiphe graminis f.sp.tritici) from the region of Slovakia and Hungary

In the year 1992 a total of 163 isolates of wheat powdery mildew were tested. The samples of mildew isolates were obtained by means of a mobile spore catching apparatus. The populations from 4 regions of Slovakia and 3 regions of Hungary were analyzed. The resistance due toPm5, Pm8 andMl-i genes at the observed locations has already been overcome. The resistance genesPm1, Pm2 and a gene combinationPm2+Pm6 ensure the protection only against a part of the patho-types of powdery mildew population. Virulence corresponding to thePm4b gene has been low so far. The regional patterns of pathogen virulence are in good agreement with the gene resistance spectrum by the cultivars grown regionally. Little differences in virulence among the populations from the regions of Slovakia and Hungary indicate that this part of Eastern Europe should be considered as an epidemiologic unit.     

Development of functional markers specific for seven Pm3 resistance alleles and their validation in the bread wheat gene pool

In the ideal case, molecular markers used for marker-assisted selection are allele-specific even if the alleles differ only by a few nucleotide polymorphisms within the coding sequence of target genes. Such ‘perfect’ markers are completely correlated with the trait of interest. In hexaploid wheat (Triticum aestivum L.) the Pm3 locus encodes seven alleles (Pm3a–Pm3g) conferring resistance to different races of Blumeria graminis f.sp. tritici, the agent of powdery mildew, a major disease of bread wheat. All Pm3 alleles are known at the molecular level. Here, we generated specific markers for the Pm3 alleles based on nucleotide polymorphisms of coding and adjacent non-coding regions. The specificity of these markers was validated in a collection of 93 modern or historically important cultivars and breeding lines of wheat and spelt (Triticum spelta L.). These markers confirmed the presence of the predicted Pm3 alleles in 31 varieties and lines known to carry Pm3 resistance alleles. In a few varieties, Pm3 alleles different from alleles previously described based on pathogenicity tests or tightly linked markers were observed. In all these cases, the identity of the marker-detected Pm3 alleles was confirmed by DNA sequence analysis. Pm3 markers confirmed the absence of known Pm3 resistance alleles in 54 European wheat and spelt varieties in which Pm3 alleles had not been previously identified. These results indicate that the developed markers are highly diagnostic for specific Pm3 resistance alleles in a wide range of varieties and breeding lines, and will be useful (1) for identifying Pm3 alleles in the wheat gene pool, (2) for efficient marker-assisted selection of these genes, and (3) for combining multiple Pm3 alleles within a single cultivar through transgenic approaches.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Rye Pm8 and wheat Pm3 are orthologous genes and show evolutionary conservation of resistance function against powdery mildew

The improvement of wheat through breeding has relied strongly on the use of genetic material from related wild and domesticated grass species. The 1RS chromosome arm from rye was introgressed into wheat and crossed into many wheat lines, as it improves yield and fungal disease resistance. Pm8 is a powdery mildew resistance gene on 1RS which, after widespread agricultural cultivation, is now widely overcome by adapted mildew races. Here we show by homology‐based cloning and subsequent physical and genetic mapping that Pm8 is the rye orthologue of the Pm3 allelic series of mildew resistance genes in wheat. The cloned gene was functionally validated as Pm8 by transient, single‐cell expression analysis and stable transformation. Sequence analysis revealed a complex mosaic of ancient haplotypes among Pm3‐ and Pm8‐like genes from different members of the Triticeae. These results show that the two genes have evolved independently after the divergence of the species 7.5 million years ago and kept their function in mildew resistance. During this long time span the co‐evolving pathogens have not overcome these genes, which is in strong contrast to the breakdown of Pm8 resistance since its introduction into commercial wheat 70 years ago. Sequence comparison revealed that evolutionary pressure acted on the same subdomains and sequence features of the two orthologous genes. This suggests that they recognize directly or indirectly the same pathogen effectors that have been conserved in the powdery mildews of wheat and rye.     

Geographical distribution of genes for resistance to formae speciales of Erysiphe graminis in common wheat

The geographical distribution of Pm10, Pm11, Pm14, and Pm15 wheat genes for resistance to inappropriate formae speciales of Erysiphe graminis was investigated using gene-for-gene relationships. Pm10 and Pm15 were very common among many indigenous accessions of common wheat collected from various areas in the world. The diversity of genotypes, which consisted of allelic combination at those loci, was high near the center of origin of common wheat and decreased with increasing distance from the center. In Europe, an apparent contrast of predominant genotypes occurred between the south and the north, suggesting that these genes are useful markers for revealing the routes by which common wheat spread in Europe. On a whole, the genes for resistance to inappropriate formae speciales were observed to be widely distributed throughout the world. We suggest that the difference between these genes and the genes for resistance to races of an appropriate forma specialis may only be in their distribution and that of their corresponding avirulence genes.     

Chromosomal location of genes for resistance to powdery mildew in common wheat (Triticum aestivum L. em Thell.) 6. Alleles at the Pm5 locus

Genetic characterization of powdery mildew resistance genes were conducted in common wheat cultivars Hope and Selpek possessing resistance gene Pm5, cvs. Ibis and Kormoran expressing resistance gene Mli, a backcross-derived line IGV 1–455 and a Triticum sphaerococcum var. rotundatum Perc. line Kolandi. Monosomic analyses revealed that one major recessive gene is located on chromosome 7B in the lines IGV 1–455 and Kolandi. Allelism tests of the F₂ and F₃ populations involving the tested resistant lines crossed with either cv. Hope or Selpek indicated that their resistance genes are alleles at the Pm5 locus. The alleles are now designated Pm5a in Hope and Selpek, Pm5b in Ibis and Kormoran, Pm5c in T. sphaerococcum var. rotundatum line Kolandi, and Pm5d in backcross-derived line IGV 1–455, respectively. Received: 5 November 1999 / Accepted: 14 April 2000     

The powdery mildew resistance gene Pm8 derived from rye is suppressed by its wheat ortholog Pm3

The powdery mildew resistance gene Pm8 derived from rye is located on a 1BL.1RS chromosome translocation in wheat. However, some wheat lines with this translocation do not show resistance to isolates of the wheat powdery mildew pathogen avirulent to Pm8 due to an unknown genetically dominant suppression mechanism. Here we show that lines with suppressed Pm8 activity contain an intact and expressed Pm8 gene. Therefore, the absence of Pm8 function in certain 1BL.1RS‐containing wheat lines is not the result of gene loss or mutation but is based on suppression. The wheat gene Pm3, an ortholog of rye Pm8, suppressed Pm8‐mediated powdery mildew resistance in lines containing Pm8 in a transient single‐cell expression assay. This result was further confirmed in transgenic lines with combined Pm8 and Pm3 transgenes. Expression analysis revealed that suppression is not the result of gene silencing, either in wheat 1BL.1RS translocation lines carrying Pm8 or in transgenic genotypes with both Pm8 and Pm3 alleles. In addition, a similar abundance of the PM8 and PM3 proteins in single or double homozygous transgenic lines suggested that a post‐translational mechanism is involved in suppression of Pm8. Co‐expression of Pm8 and Pm3 genes in Nicotiana benthamiana leaves followed by co‐immunoprecipitation analysis showed that the two proteins interact. Therefore, the formation of a heteromeric protein complex might result in inefficient or absent signal transmission for the defense reaction. These data provide a molecular explanation for the suppression of resistance genes in certain genetic backgrounds and suggest ways to circumvent it in future plant breeding.     

Mapping of five resistance genes to sugar-beet powdery mildew using AFLP and anchored SNP markers

Sugar-beet powdery mildew, caused by the fungus Erysiphe betae, now occurs in all sugar-beet growing areas and can reduce sugar yield by up to 30%. Powdery mildew resistant plants from three novel sources were crossed with sugar beet to generate segregating populations. Evaluation of resistance was carried out in artificially inoculated field and controlled environment tests. The resistance level in two of the sources was found to be significantly higher than that in currently available sugar-beet cultivars. AFLP analysis was used in combination with bulked segregant analysis to develop markers linked to the resistant phenotype in each population. Five dominant major resistance genes were identified and assigned the proposed symbols Pm2 to Pm6. Pm3 conferred complete resistance to powdery mildew; the other genes conferred high levels of partial resistance. From the use of anchoring SNP markers, two genes were located to chromosome II and three to chromosome IV. Two of the genes on chromosome IV mapped to the same location and one of the genes on chromosome II mapped to the same region as the previously identified Pm1 gene. With the availability of these genes there is now excellent potential for achieving durable resistance to sugar-beet powdery mildew, thus reducing or obviating the need for chemical control.     

<Emphasis Type="Italic">Pm37</Emphasis>, a new broadly effective powdery mildew resistance gene from <Emphasis Type="Italic">Triticum </Emphasis><Emphasis Type="Italic">timopheevii</Emphasis>

Powdery mildew is an important foliar disease in wheat, especially in areas with a cool or maritime climate. A dominant powdery mildew resistance gene transferred to the hexaploid germplasm line NC99BGTAG11 from T. timopheevii subsp. armeniacum was mapped distally on the long arm of chromosome 7A. Differential reactions were observed between the resistance gene in NC99BGTAG11 and the alleles of the Pm1 locus that is also located on chromosome arm 7AL. Observed segregation in F_2:3 lines from the cross NC99BGTAG11 × Axminster (Pm1a) demonstrate that germplasm line NC99BGTAG11 carries a novel powdery mildew resistance gene, which is now designated as Pm37. This new gene is highly effective against all powdery mildew isolates tested so far. Analyses of the population with molecular markers indicate that Pm37 is located 16 cM proximal to the Pm1 complex. Simple sequence repeat (SSR) markers Xgwm332 and Xwmc790 were located 0.5 cM proximal and distal, respectively, to Pm37. In order to identify new markers in the region, wheat expressed sequence tags (ESTs) located in the distal 10% of 7AL that were orthologous to sequences from chromosome 6 of rice were targeted. The two new EST-derived STS markers were located distal to Pm37 and one marker was closely linked to the Pm1a region. These new markers can be used in marker-assisted selection schemes to develop wheat cultivars with pyramids of powdery mildew resistance genes, including combinations of Pm37 in coupling linkage with alleles of the Pm1 locus.     

Intragenic allele pyramiding combines different specificities of wheat Pm3 resistance alleles

Some plant resistance genes occur as allelic series, with each member conferring specific resistance against a subset of pathogen races. In wheat, there are 17 alleles of the Pm3 gene. They encode nucleotide‐binding (NB‐ARC) and leucine‐rich‐repeat (LRR) domain proteins, which mediate resistance to distinct race spectra of powdery mildew. It is not known if specificities from different alleles can be combined to create resistance genes with broader specificity. Here, we used an approach based on avirulence analysis of pathogen populations to characterize the molecular basis of Pm3 recognition spectra. A large survey of mildew races for avirulence on the Pm3 alleles revealed that Pm3a has a resistance spectrum that completely contains that of Pm3f, but also extends towards additional races. The same is true for the Pm3b and Pm3c gene pair. The molecular analysis of these allelic pairs revealed a role of the NB‐ARC protein domain in the efficiency of effector‐dependent resistance. Analysis of the wild‐type and chimeric Pm3 alleles identified single residues in the C‐terminal LRR motifs as the main determinant of allele specificity. Variable residues of the N‐terminal LRRs are necessary, but not sufficient, to confer resistance specificity. Based on these data, we constructed a chimeric Pm3 gene by intragenic allele pyramiding of Pm3d and Pm3e that showed the combined resistance specificity and, thus, a broader recognition spectrum compared with the parental alleles. Our findings support a model of stepwise evolution of Pm3 recognition specificities.     

A “one-marker-for-two-genes” approach for efficient molecular discrimination of Pm12 and Pm21 conferring resistance to powdery mildew in wheat

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most important wheat diseases worldwide. Pyramiding different resistance genes into single cultivar has been proposed as one remedy to provide durable resistance. Powdery mildew resistance genes Pm12 (T6BS-6SS.6SL), transferred from Aegilops speltoides to wheat cv. Wembley, and Pm21 (T6VS.6AL), introduced from Dasypyrum villosum to wheat cv. Yangmai5, conferred broad-spectrum resistance to B. graminis f. sp. tritici. Both Pm12 and Pm21 genes are located on the short arms of homologous group six involved translocated chromosomes 6SS.6BL and 6VS.6AL, respectively. Simple sequence repeat motifs of wheat simple sequence repeat (SSR) and expressed sequence tag (EST) sequences on the short arm of homologous group six chromosomes were analyzed to develop molecular markers for discriminating chromosome arms 6AS, 6BS, 6DS, 6VS, and 6SS. One EST–SSR marker, Xcau127, was polymorphic, and therefore can be used to distinguish the two resistance genes and the respective susceptible alleles. This marker allowed us to develop an efficient “one-marker-for-two-genes” procedure for identifying powdery mildew resistance genes Pm12 and Pm21 for marker-assisted selection and gene pyramiding in wheat breeding programs. Wei Song and Chaojie Xie contributed equally to this work     

Chromosomal location of genes for resistance to powdery mildew in common wheat (Triticum aestivum L. em Thell.). 5. Alleles at the Pm1 locus

 The chromosomal location and genetic characterization of powdery mildew resistance genes were determined in the common wheat lines MocZlatka, Weihenstephan Stamm M1N and in a resistant line of Triticum aestivum ssp. spelta var. duhamelianum. Monosomic analyses revealed that one major dominant gene is located on chromosome 7A in each of the lines tested. Allelism tests with Pm1 in the backcross-derived line Axminster/8^*Cc on 7A indicated that the resistance genes are alleles at the Pm1 locus. These alleles are now designated Pm1a in line Axminster/8^*Cc, Pm1b in MocZlatka, Pm1c in Weihenstephan Stamm M1N, and Pm1d in T. spelta var. duhamelianum, respectively. Received: 10 November 1997 / Accepted: 29 January 1998     

Development of wheat near-isogenic lines for powdery mildew resistance

Using three Chinese wheat cultivars, Bainong 3217, Beijing 837 and Laizhou 953, as recurrent parents, 33 near-isogenic lines (NILs) carrying 22 powdery mildew resistance genes (Pm1c, Pm2, Pm4b, Pm12, Pm13, Pm16, Pm20, Pm21, Pm23, and 13 undocumented genes) were developed. All NILs had no significant difference to their recurrent parents in the investigated traits of agronomic importance. The results of AFLP analysis indicated Jaccards genetic similarity of the NILs with their recurrent parents varied from 0.96 to 0.98, and confirmed that the NILs had high genetic similarity with their recurrent parents. The resistance to powdery mildew was stably expressed by the relevant NILs. Eleven of the NILs were tested using molecular markers linked to the resistance genes Pm1c, Pm4b, Pm13, Pm21, PmP, PmE, PmPS5A, PmPS5B, PmY39, PmY150, and PmH, and they were all found to carry the targeted genes. The potential application of these NILs in gene discovery is discussed.     

Genetic mapping of two powdery mildew resistance genes in einkorn (Triticum monococcum L.) accessions

Powdery mildew is a severe foliar disease for wheat and could cause great yield loss in epidemic years. To explore new powdery mildew resistance genes, two einkorn accessions including TA2033 and M80, both resistant to this disease, were studied for the inheritance of resistance. Each accession possessed a single but different dominant resistance gene that was designated as Mlm2033 and Mlm80, respectively. Marker mapping indicated that they are both linked to Xgwm344 on the long arm of chromosome 7A. To establish their genetic relationship with Pm1 on 7AL, five RFLP markers previously reported to co-segregate with Pm1a were converted to STS markers. Three of them detected polymorphism between the mapping parents and were mapped close to Mlm2033 or Mlm80 or both. Xmag2185, the locus determined by the STS marker derived from PSR680, one of the RFLP markers, was placed less than 2 cM away from them. The allelism test indicated that Mlm2033 and Mlm80 are likely allelic to each other. In addition, through comparative and EST mapping, more markers linked to these two genes were identified. The high density mapping of Mlm2033 and Mlm80 will contribute to map-based cloning of the Pm1 locus. The markers for both genes will also facilitate their transfer to wheat.     

Rye-derived powdery mildew resistance gene <Emphasis Type="Italic">Pm8</Emphasis> in wheat is suppressed by the <Emphasis Type="Italic">Pm3</Emphasis> locus

Genetic suppression of disease resistance is occasionally observed in hexaploid wheat or in its interspecific crosses. The phenotypic effects of genes moved to wheat from relatives with lower ploidy are often smaller than in the original sources, suggesting the presence of modifiers or partial inhibitors in wheat, especially dilution effects caused by possible variation at orthologous loci. However, there is little current understanding of the underlying genetics of suppression. The discovery of suppression in some wheat genotypes of the cereal rye chromosome 1RS-derived gene Pm8 for powdery mildew resistance offered an opportunity for analysis. A single gene for suppression was identified at or near the closely linked storage protein genes Gli-A1 and Glu-A3, which are also closely associated with the Pm3 locus on chromosome 1AS. The Pm3 locus is a complex of expressed alleles and pseudogenes embedded among Glu-A3 repeats. In the current report, we explain why earlier work indicated that the mildew suppressor was closely associated with specific Gli-A1 and Glu-A3 alleles, and predict that suppression of Pm8 involves translated gene products from the Pm3 locus.     

Transgenic Pm3b wheat lines show resistance to powdery mildew in the field

Plant resistance (R) genes are highly effective in protecting plants against diseases, but pathogens can overcome such genes relatively easily by adaptation. Consequently, in many cases R genes do not confer durable resistance in agricultural environments. One possible strategy to make the use of R genes more sustainable depends on the modification of R genes followed by transformation. To test a possible transgenic use of R genes, we overexpressed in wheat the Pm3b resistance gene against powdery mildew under control of the maize ubiquitin promoter. Four independent transgenic lines were tested in the greenhouse and the field during 3 years. The four lines showed a five‐ to 600‐fold transgene overexpression compared with the expression of the endogenous Pm3b gene in the landrace ‘Chul’. Powdery mildew resistance was significantly improved in all lines in the greenhouse and the field, both with naturally occurring infection or after artificial inoculation. Under controlled environmental conditions, the line with the strongest overexpression of the Pm3b gene showed a dramatic increase in resistance to powdery mildew isolates that are virulent on the endogenous Pm3b. Under a variety of field conditions, but never in the greenhouse, three of the four transgenic lines showed pleiotropic effects on spike and leaf morphology. The highest overexpressing line had the strongest side effects, suggesting a correlation between expression level and phenotypic changes. These results demonstrate that the successful transgenic use of R genes critically depends on achieving an optimal level of their expression, possibly in a tissue‐specific way.     

Identification and development of diagnostic markers for a powdery mildew resistance gene on chromosome 2R of Chinese rye cultivar Jingzhouheimai

Chinese rye cultivar Jingzhouheimai (Secale cereale L.) shows a high level of resistance to powdery mildew. Identification, location, and mapping of the resistance gene would be helpful for developing a highly resistant germplasm or cultivar in wheat. Using sequential C-banding, GISH, and marker analysis, an addition chromosome with powdery mildew resistance was identified in a line derived from a cross between Chinese wheat landrace Huixianhong and rye cultivar Jingzhouheimai. The line, designated H-J DA2RDS1R(1D), had 44 chromosomes including two pairs of rye chromosomes, 1R and 2R, and lacked a pair of wheat chromosomes 1D, that is, it is a double disomic addition disomic substitution line. According to its reaction to different isolates of the powdery mildew pathogen, the resistance gene in H-J DA2RDS1R(1D) differed from the Pm8 and Pm7 genes located earlier on rye chromosomes 1R and 2R, respectively. In order to determine the location of the resistance gene, line H-J DA2RDS1R(1D) was crossed with wheat landrace Huixianhong and the F2 population and corresponding F2:3 families were tested for disease reaction and assessed with molecular markers. The results showed that a resistance gene, designated PmJZHM2RL, is located in rye chromosome arm 2RL.