Powdery Mildew Resistance Gene Research Articles

Two functional CC-NBS-LRR proteins from rye chromosome 6RS confer differential age-related powdery mildew resistance to wheat.

Rye (Secale cereale), a valuable relative of wheat, contains abundant powdery mildew resistance (Pm) genes. Using physical mapping, transcriptome sequencing, barley stripe mosaic virus-induced gene silencing, ethyl methane sulfonate mutagenesis, and stable transformation, we isolated and validated two coiled-coil, nucleotide-binding site and leucine-rich repeat (CC-NBS-LRR) alleles, PmTR1 and PmTR3, located on rye chromosome 6RS from different triticale lines. PmTR1 confers age-related resistance starting from the three-leaf stage, whereas its allele, PmTR3, confers typical all-stage resistance, which may be associated with their differential gene expression patterns. Overexpression in Nicotiana benthamiana showed that the CC, CC-NBS, and CC-LRR fragments of PMTR1 induce cell death, whereas in PMTR3 the CC and full-length fragments perform this function. Luciferase complementation imaging and pull-down assays revealed distinct interaction activities between the CC and NBS fragments. Our study elucidates two novel rye-derived Pm genes and their derivative germplasm resources and provides novel insights into the mechanism of age-related resistance, which can aid the improvement of resistance against wheat powdery mildew.

Molecular screening of squash and patisson squash collection samples using markers of the <i>Pm-0</i> gene, which controls resistance to powdery mildew

Powdery mildew (PM) is one of the most harmful diseases of cucurbits. Modern commercial varieties of squash Cucurbita pepo L. var. giraumonas Duch and patisson C. pepo var. melopepo L. received powdery mildew resistance genes from wild species. The Pm-0 resistance gene belongs to the linkage group 10; two CAPS markers were developed for its mapping [15]. The main intragenic marker NBS_S9_1495924/HaeIII is localized in the NBS-LRR region, the additional marker S9_1539675/MspI shows complete co-segregation with resistance to PM [15]. In the present study, these markers were used for molecular screening of an experimental set of squash and patisson samples (differ in resistance to powdery mildew) from the VIR gene bank collection and perspective breeding lines of the Krymsk Experiment Breeding Station branch of VIR. In total, 80 samples were investigated. Samples, carrying fragments of both CAPS markers (17) and fragments of only one marker (31) were found. Presence of two markers of the Pm-0 gene (NBS_S9_1495924/HaeIII и S9_1539675/MspI) has the strong correlation with resistance to PM (r = 0.837).
 For three powdery mildew-resistant samples, which has demonstrated presence of both markers of the Pm-0 gene, additional analysis of individual plants was done. Resistant plants with Pm-0 gene markers were self-pollinated to create resistant lines. Thus, molecular screening allowed to preserve the valuable trait of resistance during maintaining of the squash collection samples.

Development of novel wheat-rye 6RS small fragment translocation lines with powdery mildew resistance and physical mapping of the resistance gene PmW6RS.

Novel wheat-rye 6RS small fragment translocation lines with powdery mildew resistance were developed, and the resistance gene PmW6RS was physically mapped onto 6RS-0.58-0.66-bin corresponding to 18.38Mb in Weining rye. Rye (Secale cereale L., RR) contains valuable genes for wheat improvement. However, most of the rye resistance genes have not been successfully used in wheat cultivars. Identification of new rye resistance genes and transfer of these genes to wheat by developing small fragment translocation lines will make these genes more usable for wheat breeding. In this study, a broad-spectrum powdery mildew resistance gene PmW6RS was localized on rye chromosome arm 6RS using a new set of wheat-rye disomic and telosomic addition lines. To further study and use PmW6RS, 164 wheat-rye 6RS translocation lines were developed by 60Coγ-ray irradiation. Seedling and adult stage powdery mildew resistance analysis showed that 106 of the translocation lines were resistant. A physical map of 6RS was constructed using the 6RS translocation and deletion lines, and PmW6RS was localized in the 6RS-0.58-0.66-bin, flanked by markers X6RS-3 and X6RS-10 corresponding to the physical interval of 50.23-68.61Mb in Weining rye genome. A total of 23 resistance-related genes were annotated. Nine markers co-segregate with the 6RS-0.58-0.66-bin, which can be used to rapidly trace the 6RS fragment carrying PmW6RS. Small fragment translocation lines with powdery mildew resistance were backcrossed with wheat cultivars, and 39 agronomically acceptable homozygous 6RS small fragment translocation lines were obtained. In conclusion, this study not only provides novel gene source and germplasms for wheat resistance breeding, but also laid a solid foundation for cloning of PmW6RS.

Development and Identification of a Wheat-Rye Breeding Line for Harmonious Improvement Between Powdery Mildew Resistance and High Yield Potential.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a devastating disease that seriously threatens wheat yield and quality. To control this disease, host resistance is the preferred measure. However, wheat breeding is a complex process with elusive exchange and recombination of the traits from their parents. Increased resistance often leads to a decline in other key traits, such as yield and quality. Developing breakthrough germplasms with harmonious powdery mildew resistance and other key breeding traits is attractive in wheat breeding. In this study, we developed an ideal wheat breeding line AL46 that pyramided its hexaploid triticale parent-derived desirable yield traits and its wheat parent-derived powdery mildew resistance gene Pm2. Sequential genomic in situ hybridization (GISH), multicolor GISH, multicolor fluorescence in situ hybridization, and molecular marker analyses revealed that AL46 was a wheat-rye T1RS·1BL translocation line. Genetic analysis combined with function marker detection and sequence alignment were used to confirm that AL46 carried the Pm2 gene. Then, we evaluated the powdery mildew resistance and comprehensive traits of AL46, and just as we designed, AL46 showed harmonious powdery mildew resistance with some key breeding traits. This study not only developed an ideal wheat germplasm resource but also provided a successful example for pyramiding breeding, which could be a promising direction for wheat improvement in the future.

Evaluation of Powdery Mildew Resistance and Agronomic Performance of Pyramided Mungbean Lines from Marker- Assisted Selection

Gene pyramiding is an effective strategy to provide a durable and high level of powdery mildew (PM) resistance in mungbean [Vigna radiata (L.) R. Wilczek]. The objectives of this study were to evaluate PM resistance levels of pyramided BC2F1 progenies derived from marker-assisted selection of KING (recurrent parent) × SUPER5 (donor parent) cross and measure their agronomic performance under fi eld condition. The results showed that the progenies with marker combinations linked to both PM resistance genes including I85420 + I42PL222, and I27R565 were more resistant to PM (average disease severity score of 3.80) than KING, check cv. CN72, and the progenies without any marker combinations (scores of 8, 8 and 6.75, respectively). While the progenies carrying only one resistance gene were identifi ed as containing either I85420 + I42PL222, or I27R565 which showed intermediate response score of 5.83. In addition, most of the pyramided lines produced higher yield per plant than KING because of their superior pods per plant, clusters per plant, and seeds per pod. In the future, these pyramided lines with two PM resistance genes can be further developed into new PM resistant varieties via marker-assisted backcross breeding or be used as potential PM resistant sources in mungbean breeding programs.

Identification and fine mapping of PmNJ3946 for powdery mildew resistance in einkorn wheat

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a destructive wheat disease. Although it can be easily overcome by deployment of resistance genes, the resistance is often quickly compromised by pathogen virulence. Thus, exploration and characterization of new resistance genes is always ongoing. Line NJ3946 derived from a cross of einkorn wheat accessions TA2032 and M389 showed resistance to powdery mildew. Inheritance analysis of an F2 population derived from a cross of NJ3946 and M389 suggested that the resistance was conferred by a dominant allele. With polymorphic markers identified through bulked segregant analysis (BSA), this gene was mapped to a novel locus on chromosome 3A, and was designated as PmNJ3946. Bulked segregant RNA-seq analysis (BSR-seq) was conducted to obtain more closely linked markers, which allowed delimitation of the PMNJ3946 locus to a 0.9 cM interval covering a physical distance of less than 1 Mb. PMNJ3946 was flanked by Xwgrc5153 and SNP-derived marker CHS21_3A008915069, and co-segregated with SNP-derived markers CHS21_3A008939814 and CHS21_3A008943175. The PmNJ3946 discovery expands the diversity of powdery mildew resistance genes and is useful for wheat breeding.

Dissection of a rapidly evolving wheat resistance gene cluster by long-read genome sequencing accelerated the cloning of Pm69

Gene cloning in repeat-rich polyploid genomes remains challenging. Here, we describe a strategy for overcoming major bottlenecks in cloning of the powdery mildew resistance gene (R-gene) Pm69 derived from tetraploid wild emmer wheat. A conventional positional cloning approach was not effective owing to suppressed recombination. Chromosome sorting was compromised by insufficient purity. A Pm69 physical map, constructed by assembling Oxford Nanopore Technology (ONT) long-read genome sequences, revealed a rapidly evolving nucleotide-binding leucine-rich repeat (NLR) R-gene cluster with structural variations. A single candidate NLR was identified by anchoring RNA sequencing reads from susceptible mutants to ONT contigs and was validated by virus-induced gene silencing. Pm69 is likely a newly evolved NLR and was discovered in only one location across the wild emmer wheat distribution range in Israel. Pm69 was successfully introgressed into cultivated wheat, and a diagnostic molecular marker was used to accelerate its deployment and pyramiding with other R-genes.

Genetics, resistance mechanism, and breeding of powdery mildew resistance in cucumbers (Cucumis sativus L.)

Cucumber is an important vegetable worldwide, and powdery mildew (PM) is a common and serious disease of cucumbers. Breeding disease-resistant cucumber varieties is the most advantageous strategy to control this disease. In recent years, exploration and identification of cucumber PM resistance genes have achieved great advancement, and many genes have been cloned and verified using different methods. However, the resistance mechanism of cucumber PM is still unclear, and many ambiguities need to be elucidated urgently. In this review, we summarized the research advances in PM resistance in cucumbers, including genetic analysis, quantitative trait locus mapping, map-based cloning, transcriptomics, mlo-mediated PM resistance, and mining of noncoding RNAs involved in resistance. Finally, the research directions and the problems that need to be solved in the future were discussed.

Metabolic and genotypic mechanisms involved in early defensive response of powdery mildew-infected wheat

Early defensive responses of powdery mildew-infected wheat were investigated in 17 Egyptian cultivars and 21 universal powdery mildew resistance genes at seedling and adult plant stages under greenhouse conditions. Genotypic responses revealed that none of the Egyptian cultivars confer complete resistance under inoculation with Blumeria graminis f. sp. tritici. Giza-168, Misr-1, Misr-2, and Giza-171 appear to have good partial resistance to the disease, while In Sakha-93, Sakha-94 and Sids-12 exhibited susceptibility. Six genes, Pm1b, Pm3a, Pm34, Pm37, NCA6, MIAG12 and NCAG13 specific all-stage resistance, six genes, Pm1a, Pm2a, Pm3b, Pm4a, Pm6, and Pm8 showed seedling susceptibility, while highest disease severities were observed in only two genes Pm6 and Pm8. Metabolic analysis of powdery mildew-infected wheat revealed a significant increase in oxidative enzyme activities (PPO, POX, CAT), Chl. a, Chl. b and carotene contents in the moderately resistant cultivars. However, increasing in electrolyte leakage was observed in susceptible cultivars. Negative correlations were recorded between powdery mildew severity and each of PPO, POX, CAT, Chl. a, Chl. b, and Car, while a positive correlation was recorded with Electrolyte leakage. Molecular assay of resistance gene Pm38 revealed gene identification in moderately resistant cultivars and was correlated with the resistance degrees. Early defense response is dependent on metabolic and genotypic mechanisms that are controlled by the expression of resistance genes.

Two pathogen loci determine Blumeria graminis f. sp. tritici virulence to wheat resistance gene Pm1a.

Blumeria graminis f. sp. tritici (Bgt) is a globally important fungal pathogen of wheat that can rapidly evolve to defeat wheat powdery mildew (Pm) resistance genes. Despite periodic regional deployment of the Pm1a resistance gene in US wheat production, Bgt strains that overcome Pm1a have been notably nonpersistent in the United States, while on other continents, they are more widely established. A genome-wide association study (GWAS) was conducted to map sequence variants associated with Pm1a virulence in 216 Bgt isolates from six countries, including the United States. A virulence variant apparently unique to Bgt isolates from the United States was detected in the previously mapped gene AvrPm1a (BgtE-5612) on Bgt chromosome 6; an invitro growth assay suggested no fitness reduction associated with this variant. A gene on Bgt chromosome 8, Bgt-51526, was shown to function as a second determinant of Pm1a virulence, and despite < 30% amino acid identity, BGT-51526 and BGTE-5612 were predicted to share > 85% of their secondary structure. A co-expression study in Nicotiana benthamiana showed that BGTE-5612 and BGT-51526 each produce a PM1A-dependent hypersensitive response. More than one member of a B. graminis effector family can be recognized by a single wheat immune receptor, and a two-gene model is necessary to explain virulence to Pm1a.

Genetic mapping of the powdery mildew resistance gene Pm7 on oat chromosome 5D

Key messageThree independent experiments with different genetic backgrounds mapped the resistance gene Pm7 in the oat genome to the distal part of the long arm of chromosome 5D.Resistance of oat to Blumeria graminis DC. f. sp. avenae is an important breeding goal in Central and Western Europe. In this study, the position of the effective and widely used resistance gene Pm7 in the oat genome was determined based on three independent experiments with different genetic backgrounds: genome-wide association mapping in a diverse set of inbred oat lines and binary phenotype mapping in two bi-parental populations. Powdery mildew resistance was assessed in the field as well as by detached leaf tests in the laboratory. Genotyping-by-sequencing was conducted to establish comprehensive genetic fingerprints for subsequent genetic mapping experiments. All three mapping approaches located the gene to the distal part of the long arm of chromosome 5D in the hexaploid oat genome sequences of OT3098 and ‘Sang.’ Markers from this region were homologous to a region of chromosome 2Ce of the C-genome species, Avena eriantha, the donor of Pm7, which appears to be the ancestral source of a translocated region on the hexaploid chromosome 5D.

Characterization and identification of the powdery mildew resistance gene in wheat breeding line ShiCG15–009

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a serious fungal disease that critically threatens the yield and quality of wheat. Utilization of host resistance is the most effective and economical method to control this disease. In our study, a wheat breeding line ShiCG15–009, released from Hebei Province, was highly resistant to powdery mildew at all stages. To dissect its genetic basis, ShiCG15–009 was crossed with the susceptible cultivar Yannong 21 to produce F1, F2 and F2:3 progenies. After genetic analysis, a single dominant gene, tentatively designated PmCG15–009, was proved to confer resistance to Bgt isolate E09. Further molecular markers analysis showed that PmCG15–009 was located on chromosome 2BL and flanked by markers XCINAU130 and XCINAU143 with the genetic distances 0.2 and 0.4 cM, respectively, corresponding to a physic interval of 705.14–723.48 Mb referred to the Chinese Spring reference genome sequence v2.1. PmCG15–009 was most likely a new gene differed from the documented Pm genes on chromosome 2BL since its different origin, genetic diversity, and physical position. To analyze and identify the candidate genes, six genes associated with disease resistance in the candidate interval were confirmed to be associated with PmCG15–009 via qRT-PCR analysis using the parents ShiCG15–009 and Yannong 21 and time-course analysis post-inoculation with Bgt isolate E09. To accelerate the transfer of PmCG15–009 using marker-assisted selection (MAS), 18 closely or co-segregated markers were evaluated and confirmed to be suitable for tracing PmCG15–009, when it was transferred into different wheat cultivars.

Identification of Key Modules and Candidate Genes for Powdery Mildew Resistance of Wheat-Agropyron cristatum Translocation Line WAT-2020-17-6 by WGCNA

As one of the serious diseases of wheat, powdery mildew (Blumeria graminis f. sp. tritici) is a long-term threat to wheat production. Therefore, it is of great significance to explore new powdery mildew-resistant genes for breeding. The wild relative species of wheat provide gene resources for resistance to powdery mildew breeding. Agropyron cristatum (2n = 4x = 28, genomes PPPP) is an important wild relative of wheat, carrying excellent genes for high yield, disease resistance, and stress resistance, which can be used for wheat improvement. To understand the molecular mechanism of powdery mildew resistance in the wheat-A. cristatum translocation line WAT2020-17-6, transcriptome sequencing was performed, and the resistance genes were analyzed by weighted gene co-expression network analysis (WGCNA). In the results, 42,845 differentially expressed genes were identified and divided into 18 modules, of which six modules were highly correlated with powdery mildew resistance. Gene ontology (GO) enrichment analysis showed that the six interested modules related to powdery mildew resistance were significantly enriched in N-methyltransferase activity, autophagy, mRNA splicing via spliceosome, chloroplast envelope, and AMP binding. The candidate hub genes of the interested modules were further identified, and their regulatory relationships were analyzed based on co-expression data. The temporal expression pattern of the 12 hub genes was verified within 96 h after powdery mildew inoculation by RT-PCR assay. In this study, we preliminarily explained the resistance mechanism of the wheat-A. cristatum translocation lines and obtained the hub candidate genes, which laid a foundation in the exploration of resistance genes in A. cristatum for powdery mildew-resistant breeding in wheat.

Stacking effective ASR and APR rust genes for multiple disease resistance in bread wheat cultivars

Rusts and powdery mildews pose serious threats to wheat and have caused substantial yield losses worldwide. Host resistance is the most economical and sustainable approach for managing such diseases. In this study, an effective leaf rust resistance gene Lr45-derived from Secale cereale L. and a linked stem rust and powdery mildew resistance gene Sr36/Pm6-derived from Triticum timopheevii were successfully pyramided. They were validated into well-adapted Indian wheat cultivars that were already carrying the APR stem rust gene Sr2/Lr27/Yr30 through marker-assisted backcross selection (MABC) following two parallel backcrossing schemes. Three efficiently linked microsatellite markers, G372185 (Lr45), Stm773-2(Sr36), and Xgwm533 (Sr2), were used to confirm introgression of these genes. Lines with resistance genes in each background showed improved agronomic traits in comparison to their recurrent parents. These lines could be used in wheat improvement programs as potentially resistant stocks for leaf, stem rusts and powdery mildew to develop new wheat cultivars.

A chromosome-scale genome assembly of Dasypyrum villosum provides insights into its application as a broad-spectrum disease resistance resource for wheat improvement

Dasypyrum villosum is one of the most valuable gene resources in wheat improvement, especially for disease resistance. The mining of favorable genes from D. villosum is frustrated by the lack of a whole genome sequence. In this study, we generated a doubled-haploid line, 91C43DH, using microspore culture and obtained a 4.05-GB high-quality, chromosome-scale genome assembly for D. villosum. The assembly contains39 727 high-confidence genes, and 85.31% of the sequences are repetitive. Two reciprocal translocation events were detected, and 7VS-4VL is a unique translocation in D. villosum. The prolamin seed storage protein-coding genes were found to be duplicated; in particular, the genes encoding low-molecular-weight glutenin at the Glu-V3 locus were significantly expanded. RNA sequencing (RNA-seq) analysis indicated that, after Blumeria graminearum f.sp tritici (Bgt) inoculation, there were more upregulated genes involved in the pattern-triggered immunity and effector-triggered immunity defense pathways in D. villosum than in Triticum urartu. MNase hypersensitive sequencing (MH-seq) identified two Bgt-inducible MH sites (MHSs), one in the promoter and one in the 3′ terminal region of the powdery mildew resistance (Pm) gene NLR1-V. Each site had two subpeaks and they were termed MHS1 (MHS1.1/1.2) and MHS2 (MHS2.1/2.2). Bgt-inducible MHS2.2 was uniquely present in D. villosum, and MHS1.1 was more inducible in D. villosum than in wheat, suggesting that MHSs may be critical for regulation of NLR1-V expression and plant defense. In summary, this study provides a valuable genome resource for functional genomics studies and wheat-D. villosum introgression breeding. The identified regulatory mechanisms may also be exploited to develop new strategies for enhancing Pm resistance by optimizing gene expression in wheat.

Chromosomal Location of Pm12-A Novel Powdery Mildew Resistance Gene from Avena sterilis.

Identification of new, effective disease resistance genes is a very important aspect of plant breeding. Also important is the precise location of individual loci and tagging them with DNA markers for marker assisted selection. The aim of the present study was identification of the molecular markers linked with Pm12, a new effective resistance gene to powdery mildew, and their location in the oat genome. The analysis was performed on 167 F2 individuals from a hybrid of Fuchs × CN67383, with the status of the locus in each individual verified by progeny test in F3. Segregation ratios confirmed the monogenic nature of resistance. Making use of the sequence data of DNA markers and the oat OT3098 v2 genome reference assembly, Pm12 is located on chromosome 7C. A comparison was also made with the reference consensus map, to which there are more reports of mapped genes to date. The mapping results suggest that Pm12 is located in the interval 103.8-111.7 cM on this map. No powdery mildew resistance locus has been identified in this region so far, suggesting that Avena sterilis CN67383 carries a novel locus offering effective resistance in oat breeding. The information included in the oat genome annotation allowed for the identification of candidate genes in the close region of the marker cluster for Pm12. This information may provide an interesting source of further analysis of the pathways of various genes in response to the stress of powdery mildew infection.

Combating powdery mildew: Advances in molecular interactions between Blumeria graminis f. sp. tritici and wheat.

Wheat powdery mildew caused by a biotrophic fungus Blumeria graminis f. sp. tritici (Bgt), is a widespread airborne disease which continues to threaten global wheat production. One of the most chemical-free and cost-effective approaches for the management of wheat powdery mildew is the exploitation of resistant cultivars. Accumulating evidence has reported that more than 100 powdery mildew resistance genes or alleles mapping to 63 different loci (Pm1-Pm68) have been identified from common wheat and its wild relatives, and only a few of them have been cloned so far. However, continuous emergence of new pathogen races with novel degrees of virulence renders wheat resistance genes ineffective. An essential breeding strategy for achieving more durable resistance is the pyramiding of resistance genes into a single genotype. The genetics of host-pathogen interactions integrated with temperature conditions and the interaction between resistance genes and their corresponding pathogen a virulence genes or other resistance genes within the wheat genome determine the expression of resistance genes. Considerable progress has been made in revealing Bgt pathogenesis mechanisms, identification of resistance genes and breeding of wheat powdery mildew resistant cultivars. A detailed understanding of the molecular interactions between wheat and Bgt will facilitate the development of novel and effective approaches for controlling powdery mildew. This review gives a succinct overview of the molecular basis of interactions between wheat and Bgt, and wheat defense mechanisms against Bgt infection. It will also unleash the unsung roles of epigenetic processes, autophagy and silicon in wheat resistance to Bgt.

Genetic analysis of resistance to powdery mildew on 7Mg chromosome of wheat–Aegilops geniculata, development and utilization of specific molecular markers

BackgroundPowdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is prevalent in the main wheat-producing regions of China, resulting in severe yield losses in recent years. Mining and utilization of resistant genes from wild relatives of wheat is the most environmentally sound measure to control disease. Aegilops geniculata Roth (2n = 2x = 28, UgUgMgMg) is an essential and valuable disease-resistance gene donor for wheat improvement as a close relative species.ResultsIn this study, to validate powdery mildew resistance locus on chromosome 7Mg, two genetic populations were constructed and through crossing wheat – Ae. geniculata 7Mg disomic addition line NA0973-5-4-1-2-9-1 and 7Mg (7 A) alien disomic substitution line W16998 with susceptible Yuanfeng175 (YF175, authorized varieties from Shaanxi province in 2005), respectively. Cytological examination, in situ hybridization (ISH), and functional molecular markers analysis revealed that the plants carrying chromosome 7Mg showed high resistance to powdery mildew in both F1 and F2 generation at the seedling stage. Besides, 84 specific markers were developed to identify the plants carrying chromosome 7Mg resistance based on the specific-locus amplified fragment sequencing (SLAF-seq) technique. Among them, four markers were selected randomly to check the reliability in F2 segregating populations derived from YF175/NA0973-5-4-1-2-9-1 and YF175/W16998. In summary, the above analysis confirmed that a dominant high powdery mildew resistance gene was located on chromosome 7Mg of Ae. geniculata.ConclusionThe results provide a basis for mapping the powdery mildew resistance gene mapping on chromosome 7Mg and specific markers for their utilization in the future.

Non-Authenticity of Spring Barley Genotypes Revealed in Gene Bank Accessions.

Plant research and breeding depends on plant genotypes; therefore, genotype authenticity of accessions is the basic requirement for users of gene banks. Surprisingly, this extremely important topic is rarely reported in the scientific community. Non-authentic are accessions that are mislabelled and undesirable genotypes of heterogeneous accessions. In barley, we try to uncover both named problems on the basis of postulated major powdery mildew resistance genes. These are diverse, environmentally stable and their use is well documented and suitable for genotype characterization. In this contribution, we postulate resistance genes in 15 varieties represented by 157 derived lines of 32 accessions originating from seven foreign gene banks and compare these findings with previous results including those 15 identically labelled varieties from our domestic gene bank. We found that 37.5% of the gene bank accessions investigated herein were heterogeneous, and at least 20.0% were mislabelled. A large-scale molecular characterisation of varieties is now being carried out, and using authentic varieties must be one of the key requirements. Therefore, accessions of each variety from a minimum of three gene banks whose identity has been verified by reliable methods should be compared before starting new experiments. These will involve molecular varietal characterisation to serve as a foundation for future plant science research and effective crop improvement.

Germplasm Screening Using DNA Markers and Genome-Wide Association Study for the Identification of Powdery Mildew Resistance Loci in Tomato.

Powdery mildew (PM), caused by Oidium spp. in tomato, is a global concern that leads to diminished yield. We aimed to evaluate previously reported DNA markers linked to powdery mildew resistance (PMR) and identify novel quantitative trait loci (QTLs) for PMR through a genome-wide association study in tomato. Sequencing analysis of the internal transcribed spacer (ITS) of a PM strain (PNU_PM) isolated from Miryang, Gyeongnam, led to its identification as Oidium neolycopersici. Thereafter, a PM bioassay was conducted for a total of 295 tomato accessions, among which 24 accessions (4 S. lycopersicum accessions and 20 accessions of seven wild species) showed high levels of resistance to PNU_PM. Subsequently, we genotyped 11 markers previously linked to PMR in 56 accessions. PMR-specific banding patterns were detected in 15/22 PMR accessions, while no such bands were observed in the powdery mildew-susceptible accessions. The genome-wide association study was performed using TASSEL and GAPIT, based on the phenotypic data of 290 accessions and 11,912 single nucleotide polymorphisms (SNPs) obtained from the Axiom® Tomato SNP Chip Array. Nine significant SNPs in chromosomes 1, 4, 6, 8, and 12, were selected and five novel QTL regions distinct from previously known PMR-QTL regions were identified. Of these QTL regions, three putative candidate genes for PMR were selected from chromosomes 4 and 8, including two nucleotide binding site-leucine rich repeat class genes and a receptor-like kinase gene, all of which have been identified previously as causative genes for PMR in several crop species. The SNPs discovered in these genes provide useful information for understanding the molecular basis of PMR and developing DNA markers for marker-assisted selection of PMR in tomato.