Powdery Mildew Resistance Gene Research Articles

Serine/threonine kinase gene Stpk-V , a key member of powdery mildew resistance gene Pm21 , confers powdery mildew resistance in wheat

Powdery mildew resistance gene Pm21, located on the chromosome 6V short arm of Haynaldia villosa and transferred to wheat as a 6VS·6AL translocation (T6VS·6AL), confers durable and broad-spectrum resistance to wheat powdery mildew. Pm21 has become a key gene resource for powdery mildew resistance breeding all over the world. In China, 12 wheat varieties containing Pm21 have been planted on more than 3.4 million hectares since 2002. Pm21 has been intractable to molecular genetic mapping because the 6VS does not pair and recombine with the 6AS. Moreover, all known accessions of H. villosa are immune to powdery mildew fungus. Pm21 is still defined by cytogenetics as a locus. In the present study, a putative serine and threonine protein kinase gene Stpk-V was cloned and characterized with an integrative strategy of molecular and cytogenetic techniques. Stpk-V is located on the Pm21 locus. The results of a single cell transient expression assay showed that Stpk-V could decrease the haustorium index dramatically. After the Stpk-V was transformed into a susceptible wheat variety Yangmai158, the characterized transgenic plants showed high and broad-spectrum powdery mildew resistance similar to T6VS·6AL. Silencing of the Stpk-V by virus-induced gene silencing in both T6VS·6AL and H. villosa resulted in their increased susceptibility. Stpk-V could be induced by Bgt and exogenous H(2)O(2), but it also mediated the increase of endogenous H(2)O(2), leading to cell death and plant resistance when the plant was attacked by Bgt.

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

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

Collinearity-based marker mining for the fine mapping of Pm6, a powdery mildew resistance gene in wheat

The genome sequences of rice (Oryza sativa L.) and Brachypodium distachyon and the comprehensive Triticeae EST (Expressed Sequence Tag) resources provide invaluable information for comparative genomics analysis. The powdery mildew resistance gene, Pm6, which was introgressed into common wheat from Triticum timopheevii, was previously mapped to the wheat chromosome bin of 2BL [fraction length (FL) 0.50-1.00] with limited DNA markers. In this study, we saturated the Pm6 locus in wheat using the collinearity-based markers by extensively exploiting these genomic resources. All wheat ESTs located in the bin 2BL FL 0.50-1.00 and their corresponding orthologous genes on rice chromosome 4 were firstly used to develop STS (Sequence Tagged Site) markers. Those identified markers that flanked the Pm6 locus were then used to identify the collinear regions in the genomes of rice and Brachypodium. Triticeae ESTs with orthologous genes in these collinear regions were further used to develop new conserved markers for the fine mapping of Pm6. Using two F(2) populations derived from crosses of IGVI-465×Prins and IGVI-466×Prins, we mapped a total of 29 markers to the Pm6 locus. Among them, 14 markers were co-segregated with Pm6 in the IGVI-466/Prins population. Comparative genome analysis showed that the collinear region of the 29 linked markers covers a~5.6-Mb region in chromosome 5L of Brachypodium and a~6.0-Mb region in chromosome 4L of rice. The marker order is conserved between rice and Brachypodium, but re-arrangements are present in wheat. Comparative mapping in the two populations showed that two conserved markers (CINAU123 and CINAU127) flanked the Pm6 locus, and an LRR-receptor-like protein kinase cluster was identified in the collinear regions of Brachypodium and rice. This putative resistance gene cluster provides a potential target site for further fine mapping and cloning of Pm6. Moreover, the newly developed conserved markers closely linked to Pm6 can be used for the marker-assisted selection (MAS) of Pm6 in wheat breeding programs.

A new powdery mildew resistance allele at the Pm4 wheat locus transferred from einkorn (Triticum monococcum)

Powdery mildew is one of the most destructive foliar diseases of wheat. A set of differential Blumeria graminis f.sp. tritici (Bgt) isolates was used to test the powdery mildew response of a Triticum monococcum-derived resistant hexaploid line, Tm27d2. Segregation analysis of 95 F2:3 lines from a Chinese Spring/Tm27d2 cross revealed that the resistance of Tm27d2 is controlled by a single dominant gene. Using monosomic analysis and a molecular mapping approach, the resistance gene was localized to the terminal end of chromosome 2AL. The linkage map of chromosome 2AL consisted of nine simple sequence repeat markers and one sequence-tagged site (STS) marker (ResPm4) indicative for the Pm4 locus. According to the differential reactions of 19 wheat cultivars/lines with known powdery mildew resistance genes to 13 Bgt isolates, Tm27d2 carried a new resistance specificity. The complete association of the resistance allele with STS marker ResPm4 indicated that it represented a new allele at the Pm4 locus. This new allele was designated Pm4d. The two flanking markers Xgwm526 and Xbarc122 closely linked to Pm4d at genetic distances of 3.4 and 1.0 cM, respectively, are present in chromosome bin 2AL1-0.85-1.00.

Powdery mildew resistance gene (Pm-AN) located in a segregation distortion region of melon LGV

Powdery mildew is one of the most important melon pathogens all over the world. So far, many genes conferring resistance to powdery mildew of melon have been described, but few of these have been finely mapped or cloned. Two F2 populations derived from Ano2 × Hami413 and Ano2 × Queen were used to map the powdery mildew resistance gene by methods of Bulked Segregation Analysis (BSA), comparative genomics and Resistance Gene Analogues (RGA) mapping. It was found that the resistance to powdery mildew in Ano2 was conferred by a dominant gene, and the gene was named Pm-AN. The genetic analysis revealed that Pm-AN located between two codominant markers RPW and MRGH63B in linkage groupV. The genetic distances between Pm-AN and these two markers were 1.4–1.8 and 1.6–2 cM. No recombination was found between Pm-AN and markers ME/E1, SRAP23. Pm-AN was located in a RGA-rich region and cosegregated with the RGA marker MRGH5 and the resistance gene Vat. Synteny analysis showed that markers in this region were collinear between melon and cucumber. Segregation distortion was found in this region using both Ano2 × Hami413 and Ano2 × Queen F2 populations, and the distortion was more distinct in Ano2 × Hami413 F2 population. The center of segregation distortion was located in the RGA rich region harboring Pm-AN.

Suppressed recombination rate in 6VS/6AL translocation region carrying the Pm21 locus introgressed from Haynaldia villosa into hexaploid wheat

Pm21 is an effective gene for powdery mildew resistance transferred from Haynaldia villosa into common wheat cultivars. No virulence against this gene has been detected so far. A set of 42 powdery mildew isolates collected in Israel and tested in the current study also revealed no virulence against this gene. Pm21 was previously reported to be located on the short arm of 6VS/6AL translocation chromosome. We constructed a high-density genetic map of chromosome 6A, consisting of 28 PCR markers and the Pm21 gene. A comparison with previously published genetic maps of wheat chromosome 6A revealed that the recombination rate in the 6VS/6AL translocation region was poor. We assume that suppressed recombination caused by the alien H. villosa genetic material is the most reasonable explanation for the tight genetic linkage and the inadequacy between the Pm21 genetic map and the Pm21 physical map of 6A. A large number of sequence-tag sites (STS) and simple sequence repeat markers, which co-segregate with or are closely linked to the Pm21 gene, and the conversion of three resistance gene analog markers into new STS markers, provide a reliable and easy-to-use molecular tool for marker-assisted selection of Pm21 in wheat breeding programs. An additional gene, Pm31, previously reported to be derived from Triticum dicoccoides, was mapped into a similar genomic location to Pm21. Screening of the parental lines and the mapping population with Pm21 diagnostic markers clearly confirmed that the donor line of Pm31 is H. villosa and not T. dicoccoides. Therefore, we conclude that Pm21 and Pm31 refer to the same gene, derived from H. villosa, and that the designation of Pm31 as a new Pm gene was erroneous.

Codominant PCR-based markers and candidate genes for powdery mildew resistance in melon (Cucumis melo L.)

Powdery mildew caused by Podosphaera xanthii is a major disease in melon crops, and races 1, 2, and 5 of this fungus are those that occur most frequently in southern Europe. The genotype TGR-1551 bears a dominant gene that provides resistance to these three races of P. xanthii. By combining bulked segregant analysis and amplified fragment length polymorphisms (AFLP), we identified eight markers linked to this dominant gene. Cloning and sequencing of the selected AFLP fragments allowed the development of six codominant PCR-based markers which mapped on the linkage group (LG) V. Sequence analysis of these markers led to the identification of two resistance-like genes, MRGH5 and MRGH63, belonging to the nucleotide binding site (NBS)-leucine-rich repeat (LRR) gene family. Quantitative trait loci (QTL) analysis detected two QTLs, Pm-R1-2 and Pm-R5, the former significantly associated with the resistance to races 1 and 2 (LOD score of 26.5 and 33.3; 53.6 and 61.9% of phenotypic variation, respectively), and the latter with resistance to race 5 (LOD score of 36.8; 65.5% of phenotypic variation), which have been found to be colocalized with the MRGH5 and MRGH63 genes, respectively. The results suggest that the cluster of NBS-LRR genes identified in LG V harbours candidate genes for resistance to races 1, 2, and 5 of P. xanthii. The evaluation of other resistant germplasm showed that the codominant markers here reported are also linked to the Pm-w resistance gene carried by the accession 'WMR-29' proving their usefulness as genotyping tools in melon breeding programmes.

The wheat Mla homologue TmMla1 exhibits an evolutionarily conserved function against powdery mildew in both wheat and barley

The race-specific barley powdery mildew (Blumeria graminis f. sp. hordei) resistance gene Mla occurs as an allelic series and encodes CC-NB-LRR type resistance proteins. Inter-generic allele mining resulted in the isolation and characterisation of an Mla homologue from diploid wheat, designated TmMla1, which shares 78% identity with barley HvMLA1 at the protein level. TmMla1 was found to be a functional resistance gene against Blumeria graminis f. sp. tritici in wheat, hereby providing an example of R gene orthologs controlling the same disease in two different species. TmMLA1 exhibits race-specific resistance activity and its N-terminal coiled-coil domain interacts with the barley transcription factor HvWRKY1. Interestingly, TmMLA1 was not functional in barley transient assays. Replacement of the TmMLA1 LRR domain with that of HvMLA1 revealed that this fusion protein conferred resistance against B. graminis f. sp. hordei isolate K1 in barley. Thus, TmMLA1 not only confers resistance in wheat but possibly also in barley against an as yet unknown barley powdery mildew race. The conservation of functional R gene orthologs over at least 12 million years is surprising given the observed rapid breakdown of Mla-based resistance against barley mildew in agricultural ecosystems. This suggests a high stability of Mla resistance in the natural environment before domestication.

Molecular and cytogenetic identification of new wheat-Dasypyrum breviaristatum additions conferring resistance to stem rust and powdery mildew.

Two cytologically stable wheat-Dasypyrum breviarisatatum addition lines, Y93-1-6-6 and Y93-1-A6-4, were identified by integrated molecular and cytogenetic techniques. C-banding and genomic in situ hybridization (GISH) showed that Y93-1-6-6 and Y93-1-A6-4 were different wheat-D. breviaristatum additions. A total of 51 markers (primer/enzyme combinations), including 6 PCR-based Landmark Unique Gene (PLUG) markers and 45 Sequence-Tagged-Site (STS) markers, were selected from 3,774 primer/enzyme combinations to further characterize these two additions. Marker haploytpes suggested that both D. breviaristatum chromosomes in Y93-1-6-6 and Y93-1-A6-4 were rearranged. Stem rust resistance screening indicated that both additions were highly resistant to race RKQQC, whereas only Y93-1-6-6 was resistant to race TTKSK (Ug99). Powdery mildew resistance screening showed that only Y93-1-6-6 was resistant. Pedigree analysis suggested that the stem rust and powdery mildew resistance of Y93-1-6-6 was derived from D. breviaristatum, indicating that the D. breviaristatum chromosomes in Y93-1-6-6 possess a new powdery mildew resistance gene(s), and new stem rust resistance gene(s). These two additions could be used as stem rust or powdery mildew resistance sources in wheat breeding programs.

Using a limited mapping strategy to identify major QTLs for resistance to grapevine powdery mildew (Erysiphe necator) and their use in marker-assisted breeding

A limited genetic mapping strategy based on simple sequence repeat (SSR) marker data was used with five grape populations segregating for powdery mildew (Erysiphe necator) resistance in an effort to develop genetic markers from multiple sources and enable the pyramiding of resistance loci. Three populations derived their resistance from Muscadinia rotundifolia ‘Magnolia’. The first population (06708) had 97 progeny and was screened with 137 SSR markers from seven chromosomes (4, 7, 9, 12, 13, 15, and 18) that have been reported to be associated with powdery or downy mildew resistance. A genetic map was constructed using the pseudo-testcross strategy and QTL analysis was carried out. Only markers from chromosome 13 and 18 were mapped in the second (04327) and third (06712) populations, which had 47 and 80 progeny, respectively. Significant QTLs for powdery mildew resistance with overlapping genomic regions were identified for different tissue types (leaf, stem, rachis, and berry) on chromosome 18, which distinguishes the resistance in ‘Magnolia’ from that present in other accessions of M. rotundifolia and controlled by the Run1 gene on chromosome 12. The ‘Magnolia’ resistance locus was termed as Run2.1. Powdery mildew resistance was also mapped in a fourth population (08391), which had 255 progeny and resistance from M. rotundifolia ‘Trayshed’. A locus accounting for 50% of the phenotypic variation mapped to chromosome 18 and was named Run2.2. This locus overlapped the region found in the ‘Magnolia’-based populations, but the allele sizes of the flanking markers were different. ‘Trayshed’ and ‘Magnolia’ shared at least one allele for 68% of the tested markers, but alleles of the other 32% of the markers were not shared indicating that the two M. rotundifolia selections were very different. The last population, 08306 with 42 progeny, derived its resistance from a selection Vitis romanetii C166-043. Genetic mapping discovered a major powdery mildew resistance locus termed Ren4 on chromosome 18, which explained 70% of the phenotypic variation in the same region of chromosome 18 found in the two M. rotundifolia resistant accessions. The mapping results indicate that powdery mildew resistance genes from different backgrounds reside on chromosome 18, and that genetic markers can be used as a powerful tool to pyramid these loci and other powdery mildew resistance loci into a single line.Electronic supplementary materialThe online version of this article (doi:10.1007/s00122-010-1511-6) contains supplementary material, which is available to authorized users.

Evidence of host‐range expansion from new powdery mildew (Blumeria graminis) infections of triticale (×Triticosecale) in France

This study aimed to determine whether powdery mildew caused by Blumeria graminis is an endemic pathogen of triticale (×Triticosecale: Triticum × Secale), emerging as a result of recent changes in its pathogenicity, or whether it is a new pathogen, possibly resulting from hybridization between ff. spp. tritici and secalis. A secondary aim was to consider breeding practices that may have favoured this emergence. Phylogenetic analyses based upon six genes revealed the close relatedness of the novel entity and the ff. spp. tritici and secalis, but the IGS marker finally grouped together the isolates collected on triticale and on wheat, supporting the scenario of a recent host‐range expansion from wheat to triticale. Pathotype analyses concluded that virulence spectra of B. graminis infecting triticale were new in comparison to those observed for other reference formae speciales, and lack of fungicide resistance in triticale isolates strengthens the hypothesis of no or little genetic exchange between wheat and triticale populations of powdery mildew. This adaptation may follow the breakdown of plant resistance genes, which are probably not very diverse in current triticale cultivars since this criterion was not considered as a major one until recent years. Moreover, the complex selection and genetics of this hybrid cereal makes it difficult to predict the transmission of powdery mildew resistance genes.

MlAB10: A Triticum turgidum Subsp. dicoccoides Derived Powdery Mildew Resistance Gene Identified in Common Wheat

Powdery mildew is an economically important disease in wheat (Triticum aestivum L.). NC97BGTAB10 contains powdery mildew resistance introgressed from wild emmer wheat (T. turgidum subsp. dicoccoides). The objectives of this research were to genetically characterize the powdery mildew resistance in NC97BGTAB10, identify SSR markers closely linked to the resistance gene, and provide evidence that the gene is novel. One hundred thirty nine F2:3 lines from the cross NC97BGTAB10/Saluda were evaluated in the field and greenhouse for reaction to powdery mildew. Resistance was controlled by a single gene. Bulked segregant analysis identified a polymorphism between the resistant and susceptible bulks on chromosome 2BL. The population was genotyped with SSR markers that were specific to chromosome 2B and a linkage map was developed. The SSR marker Xwmc445 mapped 7 cM proximal to the resistance gene and no marker could be mapped distal to the gene. It was concluded that the resistance gene lies in the distal tip of chromosome 2BL. The powdery mildew resistance genes Pm33 and MlZec1 have also been mapped to the region. A detached leaf test with two powdery mildew isolates indicated the resistance gene in NC97BGTAB10 was different from MlZec1. No allelism test could be conducted to determine the relationships between the resistance genes in NC97BGTAB10, Pm33 or MlZec1 at this time. Thus, the resistance gene in NC97BGTAB10 was given the temporary designation MlAB10.

Identification of disease resistances in wheat-Leymus multicaulis derivatives and characterization of L. multicaulis chromatin using microsatellite DNA markers

To identify resistance to Fusarium head blight (FHB), cereal yellow dwarf virus (CYDV), stem rust (Sr), and powdery mildew (Pm) in 24 common wheat (Triticum aestivum)-Leymus multicaulis addition/translocation lines that were developed cytogenetically and to verify the authenticity of these lines using microsatellite (SSR) DNA markers. Resistance to FHB was identified in the wheat-L. multicaulis addition lines, Line 9 and Line 26, which both contained L. multicaulis-specific fragments as shown by SSR markers. The translocation line, Trans 1, and the addition lines, Line 5 and Line 29, have resistance to stem rust (IT 0). Resistance to CYDV was evaluated based on virus titers measured by enzyme linked immunosorbent assay (ELISA). The addition line, Line 23, showed low virus titer (0.15), indicating resistance to CYDV. The segregation distribution of CYDV resistance in 98 F2 plants of Line 23/CS showed a significant deviation from 3:1. Inoculation with a set of 14 differential Blumeria graminis f. sp. tritici (Bgt) isolates did not detect powdery mildew resistance in translocation line Trans 1, addition line Line 9 and the amphiploid of wheat-L. multicaulis. However, Line 26 exhibited the resistance response pattern of Kavkaz, which carries Pm8, indicating that Line 26 most likely has the powdery mildew resistance gene Pm8 inherited from its parent lines Feng Kang 7 or Feng Kang 10. Twelve SSR markers, distributed on different homeologous chromosome groups of wheat, which distinguished L. multicaulis addition/translocation chromosomes, were used to verify the presence of L. multicaulis chromatin in the putative wheat-L. multicaulis addition/translocation lines. Of the 24 addition/translocation lines investigated using the 12 polymorphic SSR markers, 18 wheat-L. multicaulis derivatives showed the expected L. multicaulis-specific fragments, indicating that all of these 18 addition/translocation lines would most likely have the introgressed L. multicaulis chromosome(s). Chromosomal rearrangements also were detected in some of the wheat-L. multicaulis introgression lines.

Marker-assisted selection for two dominant powdery mildew resistance genes introgressed into a hybrid grape population

Molecular marker-assisted selection (MAS) was carried out to track the inheritance of Run1 and Ren1, two dominant powdery mildew resistance genes that originate from different genetic resources. The Run1 locus was introgressed from a Muscadinia rotundifolia × Vitis vinifera BC 4 hybrid plant derived from a recurrent pseudo-backcross breeding scheme, whereas the Ren1 locus was introgressed from the powdery mildew-resistant V. vinifera L. variety Kishmish vatkana. Introduction of the Ren1 resistance gene of V. vinifera origin into the breeding program makes possible the long-term defence of the dominant Run1 gene. Using a BC 5 (BC 4 × Kishmish vatkana) hybrid progeny consisting of 441 plants and applying several SSR and BAC-derived CB markers, we demonstrated that the powdery mildew-resistant phenotype co-segregated with the presence of at least one resistance locus-linked marker in the genome. Our data also corroborated earlier findings that the M. rotundifolia-derived Rpv1 and Run1 loci are closely linked. To further streamline the selection process, we developed a multiplex PCR- and agarose gel electrophoresis-based method for the simultaneous detection of both Run1 and Ren1. The results illustrate that MAS offers a rapid and accurate method to select hybrid genotypes that combine multiple loci of interest in grape.

Polymorphic change of appressoria by the tomato powdery mildew Oidium neolycopersici on host tomato leaves reflects multiple unsuccessful penetration attempts

The appressorial shapes of the powdery mildews are an important clue to the taxonomy of the powdery mildew fungi, but the conidia of the tomato powdery mildew Oidium neolycopersici KTP-01 develop non-lobed, nipple-shaped, and moderately lobed or multilobed appressoria on the same leaves. To remove this ambiguity, we performed consecutive observations of sequential appressorial development of KTP-01 conidia with a high-fidelity digital microscope. Highly germinative conidia of KTP-01, collected from conidial pseudochains formed on the tomato leaves, were inoculated into host tomato and nonhost barley leaves or an artificial hydrophobic membrane (Parafilm). Events from germination initiation to appressorium formation were synchronous in all conidia on all materials used for inoculation, but post-appressorial behaviors varied among the materials. Appressoria on the membrane-stuck glass slide formed several projections at different portions of the appressoria to repeat unsuccessful penetration attempts. Similar unsuccessful penetration behavior by KTP-01 conidia was observed in the inoculations into leaves of barley plants, wild tomato species Lycopersicon peruvianum LA2172 (carrying the Ol-4 gene for powdery mildew resistance), and a susceptible host tomato ( Lycopersicon esculentum) that had been inoculated with the barley powdery mildew ( Blumeria graminis f. sp. hordei, race 1) conidia. On the barley leaves, all penetrations of KTP-01 were impeded by the papillae formed beneath the sites of the appressorial projections. On both the wild tomato and the race 1-inoculated cultivated tomato plants, KTP-01 conidia were prevented from forming functional haustoria by hypersensitive epidermal cell death; this hypersensitive reaction involved the Ol-4 gene in the wild tomato plants or the ‘induced resistance’ acquired by the nonpathogenic conidia previously inoculated into the cultivated tomato plants. All these KTP-01 conidia produced several projections on the appressoria during the repeated unsuccessful penetration attempts and eventually exhibited multilobed appressoria. On the host tomato leaves inoculated singly with KTP-01 conidia, fewer than 20 % of the conidia located appressoria on the central part of target epidermal cells and succeeded in forming functional haustoria at the first penetration attempt without forming an appressorial projection. These conidia exhibited non-lobed appressoria. The remaining conidia, however, whose appressoria were located on/near the border of the target epidermal cells, were more likely to fail to penetrate at the first penetration, and then to develop additional projections for subsequent penetrations. Most conidia succeeded in forming functional haustoria at the second to fourth penetration attempts, but a few conidia failed to produce haustoria at all attempted penetrations. Eventually, the conidia that succeeded at the second penetration possessed a single appressorial projection (exhibiting the nipple-shaped appressoria), whereas the remaining conidia exhibited moderately lobed appressoria with two to four appressorial projections and multilobed appressoria, with more projections. Thus, the present study revealed that the basic shape of appressoria of KTP-01 was the non-lobed type, and that polymorphic changes of the appressoria occurred as a result of successive production of projections during repeated unsuccessful penetration attempts.

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 F(2) segregating population and F(3) 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.

CONSTRUCTION OF A GENETIC LINKAGE MAP AND IDENTIFICATION OF A MAJOR QTL CONFERRING RESISTANCE TO POWDERY MILDEW IN THE MELON GENOTYPE TGR-1551

Melon (Cucumis melo L.) is a valuable crop grown in temperate and tropical regions worldwide. Powdery mildew caused by Podosphaera xanthii is one of the most frequent diseases of melon. The genotype TGR-1551 is resistant to races 1, 2, and 5 of powdery mildew. We constructed a genetic linkage map using an F 2 population derived from a cross between TGR-1551 and the susceptible Spanish cultivar 'Bola de Oro'. The software JoinMap ® 4.0 was used to locate on the linkage map a total of 429 loci: 241 AFLP, 55 RAPD, 117 SSR, 14 SCAR/CAPS/dCAPS, and 2 phenotypic traits (Vat and a loci), which were distributed among 12 linkage groups with a LOD score ≥6.0. The map spans 1318.2 cM, with an average of 3.1 cM/ marker. Powdery mildew resistance was evaluated as a quantitative trait and then, QTL analyses were performed using interval mapping and multiple QTL model mapping with MapQTL ® 5.0 software. Both methods detected a dominant QTL for resistance to either 1, 2 or 5 races of powdery mildew, all located at the same genomic region of G4. Therefore, these QTL were considered as a unique QTL designated as Pm-R. This QTL was supported by LOD scores of 26.5, 33.3, and 36.2, explaining 53.6, 61.9, and 64.9% of the phenotypic variance in the mapping population for resistance to powdery mildew races 1, 2, and 5, respectively. In addition, four codominant markers, previously described as linked to the dominant powdery mildew resistance gene, were linked to the Pm-R QTL, which suggests the correspondence between this QTL and the dominant resistance gene. One of those codominant markers, PM3-CAPS, co-segregated with the Pm-R QTL, and the markers PM2-CAPS and PM4-dCAPS flanked this QTL. These markers could be used to select resistance to powdery mildew in melon breeding.

Identification of differentially expressed genes between powdery mildew resistant near-isogenic line and susceptible line of cucumber by suppression subtractive hybridization

A population of BC 7 near-isogenic cucumber line (NIL210) was made from a cross between the powdery mildew susceptible cultivar (D8) and resistant cultivar (JIN5-508) in order to get powdery mildew resistance genes in cucumber. In this study, suppression subtractive hybridizations (SSHs) were applied to identify differential expressed genes between NIL210 and D8. 166 differentially expressed ESTs were identified from two SSH cDNA libraries including constitutively expressed resistant and susceptible factors. Most of the classified genes belong to four pathogenesis-related categories: ‘disease/defense’, ‘signal transduction’, ‘transcription’ and ‘transporter’, which altogether accounted for 46–53% of the genes classified. Some candidate powdery mildew resistant genes were also identified, such as GT053536 (TIR-NBS-LRR protein) and GT053570 (phenylalanine ammonia-lyase 2). These results will contribute to the better understanding of molecular mechanisms underlying powdery mildew resistance in cucumber.

SCREENING OF DAMASK ROSE GENOTYPES FOR POWDERY MILDEW RESISTANCE

Podosphaera pannosa is one of the most common diseases that attacks both garden and greenhouse roses worldwide. It is also an important disease of Rosa damascena in Iran. Since Iran appears to be a center of genetic diversity for Damask rose, it is expected that powdery mildew resistance genes would be found. Therefore, a set of Damask roses from diverse regions of Iran (Fars, Isfahan, Kerman and East Azerbaijan provinces) were screened for powdery mildew resistance. The plants were screened for natural infection over two years (2006 and 2007) both in the field and in pots. The diseases index and factor were calculated for each genotype and severity of disease was compared at different scoring times. Screening in the field was not consistent, probably due to non-uniform pathogen distribution, but potted plants that were placed in favorable environmental conditions (half-shade with relatively high humidity) gave consistent ratings over replications. Significant differences in the rate of powdery mildew development were found between genotypes. The East Azerbaijan (colder with higher rainfall and humidity) and Kerman (warmer and dryer region) genotypes were recognized as the most resistant and susceptible genotypes respectively. This is the first report on resistance for powdery mildew in Damask roses. Results are promising for planning the future breeding programs on this valuable species.

Isolation of resistance gene analogues to powdery mildew resistance sequences in hexaploid wheat

This paper reports the characterization of the powdery mildew resistance homologous genes family of Triticum aestivum. Using degenerate primer pair for wheat resistance genes, we have cloned seven 3' truncated powdery mildew resistance gene homologous fragments Tpc5a, Tp25a, Tp25b, Tp3a5a, Tp3a5b, Tp4b5a and Tp4b5b. These fragments were sequenced. The deduced amino acid sequences showed that six of them have premature stop codons. All these sequences had a very high level of similarity to known Pm resistance genes such as Pm3a, Pm3b, Pm3d and pm3f in hexaploid wheat. By ignoring the stop codons in the sequences, their deduced protein sequences were of coiled-coil (CC)-nucleotide binding site (NBS)-leucine repeat rich (LRR) structure. These results suggest that there are many powdery mildew resistance gene analogues in both resistant and susceptible wheat. Among them, small insertion/deletion events and point mutations can result in the diversity of wheat Pm resistance homologous genes.