Genetical and RFLP studies at the Mla locus conferring powdery mildew resistance in barley

Summary The complex structure of the multigene family at the Mla locus conferring powdery mildew resistance in barley was studied by making diallel crosses between several near-isogenic lines carrying different Mla alleles. The mode of inheritance of the Mla alleles investigated was determined to be dominant for Mla1, Mla6, Mla7 and Mla13 and semidominant for Mla3, Mla12 and Mla20. F₁ plants were backcrossed to the susceptible recurrent parent in order to identify susceptible and double-resistant recombinants in the BC₁F₁ generation. Out of 17605 progenies tested in the BC₁F₁ generation, two susceptible recombinants, one between Mla1 and Mla12 and one between Mla13 and Mla20 were confirmed. The former was also verified by RFLP analysis.     

Use of RFLP markers for the identification of alleles of the Pm3 locus conferring powdery mildew resistance in wheat (Triticum aestivum L.)

The objective of this study was to identify molecular markers linked to genes for resistance to powdery mildew (Pm) in wheat using a series of Chancellor near-isogenic-lines (NILs), each having one powdery mildew resistance gene. A total of 210 probes were screened for their ability to detect polymorphism between the NILs and the recurrent parent. One of these restriction fragment length polymorphism (RFLP) markers (Xwhs179) revealed polymorphism not only between the NILs for the Pm3 locus, but also among NILs possessing different alleles of the Pm3 locus. The location of the marker Xwhs179 was confirmed to be on homoeologous chromosome group 1 with the help of nullitetrasomic wheat lines. The linkage relationship between this probe and the Pm3 locus was estimated with double haploid lines derived from a cross between wheat cvs Club and Chul (Pm3b). The genetic distance was determined to be 3.3±1.9 cM.     

Recombination of alleles conferring specific resistance to powdery mildew at the Mla locus in barley.

In barley (Hordeum vulgare L.), the Mla locus conditions reaction to the powdery mildew fungus Erysiphe graminis f.sp. hordei. Enrichment for genetic recombinants in the Mla region is possible by screening for recombination events between the flanking endosperm storage proteins hordeins C and B. Reciprocal crosses were made between the Franger (C.I. 16151) and Rupee (C.I. 16155) lines carrying the (Mla6 + Mla14) and Mla13 alleles, respectively. Recombinants were identified from F2 segregants by analyzing the extracted hordein polypeptides by sodium dodecyl sulphate - polyacrylamide gel electrophoresis. Two hundred and seventy-six recombinant gametes were identified from the 1800 seeds that were screened. Recombination of Mla alleles was analyzed by inoculating F4 recombinant lines with three isolates of E. graminis (A27, 5874, and CR3), which recognize specific Mla alleles. The linkage order established is Hor1-Mla6-Mla13-Mla14-Hor2. The genetic distances between Hor1-Mla6, Mla6-Mla13, and Mla13-Hor2, obtained using Mapmaker 3.0b F3 intercross analysis, are 3.9, 0.2, and 5.2 cM, respectively.     

A high-resolution genetic map and a diagnostic RFLP marker for the Mlg resistance locus to powdery mildew in barley

The semi-dominantly acting Mlg resistance locus in barley confers race-specific resistance to the obligate biotrophic fungus Erysiphe graminis f.sp. hordei. A high-resolution genetic map was constructed at Mlg based on a cross between the near-isogenic barley lines Pallas BC₅ Mlg and Pallas mlg. A total of 2000 F₂ progeny were inspected by cleaved amplified polymorphic sequence (CAPS) analysis, defining a 4.47 cM interval encompassing the resistance locus. Pathogen challenge of the segregants with multiple powdery mildew isolates uncovered a novel resistance specificity in Pallas BC₅ Mlg. Probes from within 4.0 cM of Mlg were mapped in rice, revealing orthologues on five different rice chromosomes and suggesting multiple breaks of chromosomal collinearity in this region between the two grass species. The most tightly Mlg-linked RFLP marker, MWG032, was shown to reliably detect the presence of the resistance allele in a collection of 30 European barley cultivars. Received: 23 March 2000 / Accepted: 20 April 2000     

Localization of the Laevigatum powdery mildew resistance gene to barley chromosome 2 by the use of RFLP markers

Summary The powdery mildew disease resistance gene Ml(La) was found to belong to a locus on barely chromosome 2. We suggest that this locus be designated MlLa. Linkage analysis was carried out on 72 chromosome-doubled, spring-type progeny lines from a cross between the winter var Vogelsanger Gold and the spring var Alf. A map of chromosome 2 spanning 119cM and flanked by two peroxidase gene loci was constructed. In addition to the Laevigatum resistance locus the map includes nine RFLP markers, the two peroxidase gene loci and the six-row locus in barley.     

Chromosomal localization and molecular-marker tagging of the powdery mildew resistance gene (Lv) in tomato

We report the tagging of a powdery mildew [Leveillula taurica (Lév.) Arnaud.] resistance gene (Lv) in tomato using RAPD and RFLP markers. DNA from a resistant (cv Laurica) and a susceptible cultivar were screened with 300 random primers that were used to amplify DNA of resistant and susceptible plants. Four primers yielded fragments that were unique to the resistant line and linked to the resistance gene in an F₂ population. One of these amplified fragments, OP248, with a molecular weight of 0.7 kb, was subsequently mapped to chromosome 12, 1 cM away from CT134. Using RFLP markers located on chromosome 12, it was shown that approximately one half of chromosome 12 (about 42 cM), in the resistant variety is comprised of foreign DNA, presumably introgressed with the resistance gene from the wild species L. chilense. Further analysis of a backcross population revealed that the Lv gene lies in the 5.5-cM interval between RFLP markers, CT211 and CT219. As a prelude to map-based cloning of the Lv gene, we are currently enriching the density of markers in this region by a combination of RAPD primers and other techniques.     

Chemical-induced resistance against powdery mildew in barley: the effects of chitosan and benzothiadiazole

Chitosan (CHT), a deacetylated chitin derivative, and benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH), a non toxic synthetic functional analogue of salicylic acid, were applied as foliar spray to barley plants (Hordeum vulgare L.), to compare their effectiveness in inducing resistance against Blumeria graminis f. sp. hordei and to investigate the underlying defence response. After an induction phase of 3 days (IP, time elapsed between treatment and fungal inoculation) both compounds reduced significantly the infection on the primary leaf, namely of 55.5% for CHT and of 68.9% for BTH, showing the induction of a good level of local resistance (LAR). A 5-day IP further reduced the infected areas in BTH treated plants (−77.2%) but not in CHT treated ones (−47.1%). Furthermore, both CHT and BTH also induced SAR, being the infection in the second non treated leaves reduced of 57% and 76.2%, respectively, as evaluated at 10-day IP. Both BTH and CHT induced oxidative burst and phenolic compound deposition in treated leaves, creating an hostile environment that slowed down the fungal spreading by impairing haustorium development. However, the greater efficacy of BTH was possibly due to: i) a greater reinforcement of papilla; ii) a higher level and the more homogeneous diffusion of H₂O₂ in the treated leaf tissues and iii) an induced hypersensitive-like response in many penetrated cells.     

Molecular marker analyses of powdery mildew resistance in barley

Powdery mildew, caused byEryisphe graminis f. sp.hordei, is one of the most important diseases of barley (Hordeum vulgare). A number of loci conditioning resistance to this disease have been reported previously. The objective of this study was to use molecular markers to identify chromosomal regions containing genes for powdery mildew resistance and to estimate the resistance effect of each locus. A set of 28 F₁ hybrids and eight parental lines from a barley diallel study was inoculated with each of five isolates ofE. graminis. The parents were surveyed for restriction fragment length polymorphisms (RFLPs) at 84 marker loci that cover about 1100 cM of the barley genome. The RFLP genotypes of the F₁s were deduced from those of the parents. A total of 27 loci, distributed on six of the seven barley chromosomes, detected significant resistance effects to at least one of the five isolates. Almost all the chromosomal regions previously reported to carry genes for powdery mildew resistance were detected, plus the possible existence of 1 additional locus on chromosome 7. The analysis indicated that additive genetic effects are the most important component in conditioning powdery mildew resistance. However, there is also a considerable amount of dominance effects at most loci, and even overdominance is likely to be present at a number of loci. These results suggest that quantitative differences are likely to exist among alleles even at loci which are considered to carry major genes for resistance, and minor effects may be prevalent in cultivars that are not known to carry major genes for resistance.     

Genes involved in resistance to powdery mildew in barley differentially modulate root colonization by the mycorrhizal fungus Glomus mosseae

 Arbuscular mycorrhizal fungi (AMF) and Erysiphe graminis are obligate biotrophic fungi with different outcomes in their interaction with plants, different targeted host tissues, but similar patterns of development and infection processes. These similarities raise the question of whether the two types of biotrophic fungal infections have common features in their regulation. To investigate this question, we compared a number of Ror and Rar barley mutants susceptible to E.graminis f. sp. hordei, as well as their resistant progenitors, for susceptibility to infection by the AMF Glomus mosseae. The two powdery mildew-resistant lines BC Ingrid and Sultan presented a similar reduction in G. mosseae development within roots when compared to the wildtype cultivar Ingrid, indicating a systemic effect of the altered genes in the plant. Ror and Rar mutants, in which susceptibility to powdery mildew is restored, showed increased resistance to AM fungal development in their roots when compared to their progenitors, which suggests that corresponding mutations must have affected genes which differentially modulate symbiotic and pathogenic biotrophic plant-fungus interactions. Accepted: 16 September 1999     

Genetic analysis of the obligate parasitic barley powdery mildew fungus based on RFLP and virulence loci

Summary Genome organization of the biotrophic barley powdery mildew fungus was studied using restriction fragment length polymorphism (RFLP). Genomic DNA clones containing either low-or multiple-copy sequences appeared to be the best RFLP markers, as they frequently revealed polymorphisms that could be readily detected. A total of 31 loci were identified using 11 genomic DNA clones as probes. Linkage analysis of the 31 RFLP loci and five virulence loci resulted in the construction of seven groups of linked loci. Two of these contained both RFLP markers and virulence genes. RFLP markers were found to be very efficient in characterizing mildew isolates, as only three markers were necessary to differentiate 28 isolates. The DNA of the barley powdery mildew fungus appeared to contain a considerable number of repetitive sequences dispersed throughout the genome.     

RFLP mapping of three new loci for resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) in barley

Three new major, race-specific, resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) were identified in three barley lines, RS42-6*O, RS137-28*E, and HSY-78*A, derived from crosses with wild barley (Hordeum vulgare ssp. spontaneum). The resistance gene origining from wild barley in line RS42-6*O, showed a recessive mode of inheritance, whereas the other wild barley genes were (semi)-dominant. RFLP mapping of these three genes was performed in segregating F₂ populations. The recessive gene in line RS42-6*O, was localized on barley chromosome 1S (7HS), while the (semi)-dominant genes in lines RS137-28*E, and HSY-78*A, were localized on chromosomes 1L (7HL) and 7L (5HL), respectively. Closely linked RFLP clones mapped at distances between 2.6cM and 5.3 cM. Hitherto, specific loci for powdery mildew resistance in barley had not been located on these chromosomes. Furthermore, tests for linkage to the unlocalized resistance gene Mlp revealed free segregation. Therefore, these genes represent new loci and new designations are suggested: mlt (RS42-6*O), Mlf (RS137-28*E), and Mlj (HSY-78*A). Comparisons with mapped QTLs for mildew resistance were made and are discussed in the context of homoeology among the genomes of barley (H-vulgare), wheat (Triticum aestivum), and rye (Secale cereale). Duplications of RFLP bands detected in the neighbourhood of Mlf and mlt might indicate an evolutionary interrelationship to the Mla locus for mildew resistance.     

AB-QTL analysis in spring barley. I. Detection of resistance genes against powdery mildew, leaf rust and scald introgressed from wild barley

The objective of this study was to map new resistance genes against powdery mildew (Blumeria graminis f. sp. hordei L.), leaf rust (Puccinia hordei L.) and scald [Rhynchosporium secalis (Oud.) J. Davis] in the advanced backcross doubled haploid (BC₂DH) population S42 derived from a cross between the spring barley cultivar Scarlett and the wild barley accession ISR42-8 (Hordeum vulgare ssp. spontaneum). Using field data of disease severity recorded in eight environments under natural infestation and genotype data of 98 SSR loci, we detected nine QTL for powdery mildew, six QTL for leaf rust resistance and three QTL for scald resistance. The presence of the exotic QTL alleles reduced disease symptoms by a maximum of 51.5, 37.6 and 16.5% for powdery mildew, leaf rust and scald, respectively. Some of the detected QTL may correspond to previously identified qualitative (i.e. Mla) and to quantitative resistance genes. Others may be newly identified resistance genes. For the majority of resistance QTL (61.0%) the wild barley contributed the favourable allele demonstrating the usefulness of wild barley in the quest for resistant cultivars.     

Identification of RFLP markers closely linked to the H1 gene conferring resistance to Globodera rostochiensis in potato

Summary Resistance to the root cyst nematode Globodera rostochiensis is an agronomic trait that is at present incorporated into most new potato varieties. Major dominant genes are available that originate from wild and cultivated Solanum species closely related to the cultivated European potato (Solanum tuberosum ssp. tuberosum). One of those genes, H1, from S. Tuberosum ssp. andigena, was mapped to a distal position on potato chromosome V using restriction fragment length polymorphism (RFLP) markers. The H1 locus segregates independently from Gro1, a second dominant gene presumably from S. Spegazzinii that confers resistance to G. Rostochiensis and which has been mapped to chromosome VII. One marker, CP113, was linked without recombination to the H1 locus.     

亚蔬中心绿豆品种(系)抗白粉病评价

采用苗期人工接种鉴定法,在大棚种植条件下对12个亚蔬中心(AVRDC)绿豆品种白粉病抗性进行了鉴定评价。结果显示,VC1560C、V4785和VC2768A三个品种高抗(HR)白粉病,VC6173-14、V1132为中抗(MR)白粉病品种。其它品种对白粉病表现高度感病。在田间种植条件下对亚蔬中心16个抗豆象回交9代品系(BC9)进行了成株期白粉病抗性鉴定。与对照感病品种 VC1973、VC1178A 相比,VC6459-3-6-37和 VC6458-6-3-16对白粉病具有一定抗性,但白粉病感染程度仍很严重,其它14个 BC9品系均对白粉病表现高度感病。     

Quantitative trait loci mapping of adult-plant resistance to powdery mildew in triticale

Powdery mildew (PM) is a common disease caused by Blumeria graminis, which affects cereals and has recently adapted to triticale. Adult-plant resistance (APR) genes provide durable protection of crops from the disease. Quantitative trait loci corresponding to the APR effects were mapped in an F₂ population of “Lamberto” (susceptible) × “Moderto” (resistant). A genetic map of winter triticale was constructed based on the segregation of 863 DArT, 38 microsatellite and 10 resistance gene analogue markers. Composite interval mapping (CIM) was applied to identify three QTLs for maximum disease severity (MDS) and two for the area under disease progress curve (AUDPC) conferring resistance to the powdery mildew on chromosomes: 6A, 7A, 1B and 4R. The 39% variation in AUDPC was explained by the main QTL localised on chromosome 4R. Genes coding TRIUR3 proteins, serine/threonine protein kinase and cell wall associated kinases were localised in silico within the QTL and alternative DNA markers were proposed for flexible use in laboratories of diversified throughput.     

The transfer of a powdery mildew resistance gene from Hordeum bulbosum L to barley (H. vulgare L.) chromosome 2 (2I)

Hordeum bulbosum L. is a source of disease resistance genes that would be worthwhile transferring to barley (H. vulgare L.). To achieve this objective, selfed seed from a tetraploid H. vulgare x H. bulbosum hybrid was irradiated. Subsequently, a powdery mildew-resistant selection of barley phenotype (81882/83) was identified among field-grown progeny. Using molecular analyses, we have established that the H. bulbosum DNA containing the powdery mildew resistance gene had been introgressed into 81882/83 and is located on chromosome 2 (2I). Resistant plants have been backcrossed to barley to remove the adverse effects of a linked factor conditioning triploid seed formation, but there remains an association between powdery mildew resistance and non-pathogenic necrotic leaf blotching. The dominant resistance gene is allelic to a gene transferred from H. bulbosum by co-workers in Germany, but non-allelic to all other known powdery mildew resistance genes in barley. We propose Mlhb as a gene symbol for this resistance.     

Sources of powdery mildew resistance in barley landraces from morocco

A total of forty eight accessions of barley landraces from Morocco were screened for resistance to powdery mildew. Twenty two (46%) of tested landraces showed resistance reactions and thirty four single plant lines were selected. Eleven of these lines were tested in seedling stage with seventeen and another twenty three lines with twenty three isolates of powdery mildew respectively. The isolates were chosen according to the virulence spectra observed on the ‘Pallas’ isolines differential set. Line 229–2–2 was identified with resistance to all prevalent in Europe powdery mildew virulence genes. Lines 230–1–1, 248–1–3 showed susceptible reaction for only one and lines 221–3–2, 227–1–1, 244–3–4 for only two isolates respectively. Three different resistance alleles (Mlat, Mla6, and MLA14) were postulated to be present in tested lines alone or in combination. In thirty (88%) tested lines it was impossible to determine which specific gene or genes for resistance were present. Most probably these lines possessed alleles not represented in the ‘Pallas’ isolines differential set. The distribution of reaction type indicated that about 71% of all reaction types observed were classified as powdery mildew resistance (scores 0, 1 and 2). Majority (79%) of resistance reaction types observed in tested lines was intermediate resistance reaction type two and twenty three lines (68%) showed this reaction for inoculation with more than 50% isolates used. The use of new effective sources of resistance from Moroccan barley landraces for diversification of resistance genes for powdery mildew in barley cultivars was discussed.     

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.     

Analysis of molecular markers associated with powdery mildew resistance genes in peach (Prunus persica (L.) Batsch)xPrunus davidiana hybrids

A progeny of 77 hybrids issued from a cross between two heterozygous Prunus, peach [P. persica (L.) Batsch] (variety Summergrand) and a related species, P. davidiana (clone 1908), was analysed for powdery mildew resistance in five independent experiments. This population was also analysed for its genotype with isoenzyme and RAPD markers in order to map the genes responsible for resistance. A genetic linkage map was generated for each parent. The Summergrand linkage map is composed of only four linkage groups including 15 RAPD markers and covering 83.1 centiMorgans (cM) of the peach nuclear genome, whereas the P. davidiana linkage map contains 84 RAPD markers and one isoenzyme assigned to ten linkage groups and covering 536 cM. Significant associations between molecular markers and powdery mildew resistance were found in each parent. For P. davidiana, one major QTL with a very strong effect and five other QTLs with minor effects were located in different linkage groups. For Summergrand, three QTLs for powdery mildew resistance, with minor effects, were also detected. Consequently, evidence is given here that the powdery mildew resistance of P. davidiana clone 1908 and P. persica variety Summergrand is not a monogenic character but is controlled by at least one major gene and several minor genes.