Search for novel genes for resistance to powdery mildew (Sphaerotheca fuliginae) in cucumber (Cucumis sativus)

Summary The current powdery mildew (Sphaerotheca fuligninea) resistant cucumber varieties suffer from leaf chlorosis during autumn, winter and early spring cultivation in the Netherlands. Therefore screening was carried out for novel powdery mildew resistance genes. From 177 accessions, derived from different sources, 108 accessions proved to be partially resistant to S. fuliginea. Crosses were made with 53 resistant accessions to distinguish the presence of novel genes. It is likely that the accessions C. sativus 2145, C. sativus LV 41, PI 188807, Vladivostokij, White and Yellow 1 have one or more recessive powdery mildew resistance genes, different from powdery mildew resistance genes of the line NPI, which was used for variety breeding. Powdery mildew resistance tests with S. fuliginea give similar results in different regions of the world.Abbreviations pmr powdery mildew resistance     

Molecular mapping of powdery mildew resistance genes in wheat: A review

Powdery mildew, caused by Blumeria graminis f. sp. tritici (syn. Erysiphe graminis f. sp. tritici), is one of the most important diseases of common wheat (Triticum aestivum L.) worldwide. Molecular mapping and cloning of genes for resistance to powdery mildew in hexaploid wheat will facilitate the study of molecular mechanisms underlying resistance to powdery mildew diseases and help understand the structure and function of powdery mildew resistance genes, and permit marker-assisted selection in breeding programs. So far, 48 genes/alleles for resistance to powdery mildew at 32 loci have been identified and located on 16 different chromosomes, of which 21 resistance genes/alleles have been tagged by restriction fragment length polymorphisms (RFLPs), random-amplified polymorphic DNAs (RAPDs), amplified fragment length polymorphisms (AFLPs), sequence characterized amplified regions (SCARs), sequence-tagged sites (STS) or simple sequence repeats (SSRs). Several quantitative trait loci (QTLs) for adult plant resistance (APR) to powdery mildew have been associated with molecular markers. The detailed information on chromosomal location and molecular mapping of these genes has been reviewed. Isolation of powdery mildew resistance genes and development of valid molecular markers for pyramiding resistance genes in breeding programs is also discussed.     

Resistance to powdery mildew in selections from Moroccan barley landraces

Seventy-five barley landraces from Morocco were tested for resistance to powdery mildew and a number of different resistance genes were detected. Thirty-five isolates of Blumeria graminis f. sp. hordei and the Pallas isoline differential set were used. Isolates used in the experiment had virulences corresponding to all major resistance genes used in Europe. Forty-four of the tested landraces showed resistant reactions. From each of these landraces, one to five resistant plants were selected and 92 single plant lines were created. Six lines selected from 3landraces were assumed to carry the mlo gene but they were discarded after microscopic investigation. Seventeen lines were tested in the seedling stage with 17isolates and another 69 lines were tested with 23 differential isolates. These lines showed 71 reaction spectra to isolates of powdery mildew. Eight lines (9%), 255-3-3, 282-3-4, 286-1-1, 294-2-3,294-2-4, 295-1-2, 308-1-2 and 327-2-1, selected from 7 landraces showed resistance to all isolates. Seventy-eight lines (90%) showed a resistant infection type 2with more than 50% of the isolates used. In most of the selected lines (86%) unknown genes, alone or in combination with known specific resistance genes, were detected. Four different resistance alleles (Mlat, Mla6, Mla14 and Mla1) were postulated to be present in the tested lines. The most common was Mlat, which was postulated in 35 (41%) lines. The use of newly identified sources of powdery mildew resistance in barley breeding is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development and characterization of a new 1BL.1RS translocation line with resistance to stripe rust and powdery mildew of wheat

The 1BL.1RS wheat-rye translocation from Petkus rye has contributed substantially to the world wheat production. However, following the breakdown of disease resistance genes in 1RS, its importance for wheat improvement decreased. We have developed a new 1BL.1RS line, R14, by means of crossing rye inbred line L155, selected from Petkus rye to several wheat cultivars. One new gene each, for stripe rust and powdery mildew resistance, located on 1RS of the line R14, are tentatively named YrCn17 and PmCn17. YrCn17 and PmCn17 confer resistance to Puccinia striiformis f. sp. tritici pathotypes that are virulent on Yr9, and Blumeria graminis f. sp. tritici pathotypes virulent on Pm8. These two new resistances, YrCn17 and PmCn17, are now available for wheat improvement programs. The present study indicates that rye cultivars may carry yet untapped variations as potential sources of resistance.     

Discovery,characterization and exploitation of Mlo powdery mildew resistance in barley

Summary Mlo resistance to barley powdery mildew is a relatively new kind of resistance. It was originally described in a powdery mildew resistant barley mutant in 1942 and has been mutagen-induced repeatedly since then. About 1970 it was also recognized in barley landraces collected in Ethiopia in the 1930s. It is unique in that 1) Mlo resistance does not conform to the gene-for-gene system; 2)mlo genes originating from different mutational events map as non-complementing recessive alleles in one locus; 3) all alleles confer the same phenotype, though with small quantitative differences; 4) it is effective against all isolates of the pathogen; and 5) the resistance is caused by rapid formation of large cell wall appositions at the encounter sites preventing penetration by the fungus. Powdery mildew isolates with elevated Mlo aggressiveness have been produced on barley in the laboratory, but have not been found in nature. Mlo resistance is considered very durable. The exploitation of Mlo resistance has been hampered by pleiotropic effects of themlo genes, vix. necrotic leaf spotting and reduced grain yield, but they have been overcome by recent breeding work. During the 1980s Mlo-resistant spring barley varieties have become cultivated extensively in several European countries, in 1990 on about 700,000 ha.     

Isolation of wheat mutants with increased resistance to powdery mildew from small induced variant populations

Small variant wheat populations created by induced mutagenesis (n = 69) or adventitious regeneration (n = 66) were intensively screened for an altered response (compared to the parent variety ‘Guardian’) to the causal pathogen of powdery mildew in wheat, Blumeria graminis f. sp. tritici. Intensive field screening following natural infection of replicated plots of wheat lines over two years revealed a total of 13 mutants exhibiting significantly greater resistance than ‘Guardian’: eight from induced mutagenesis (11.6%) of the M2 population and five from adventitious regeneration (7.6%). Complete resistance was identified in two lines, (one (M66) developed following induced mutagenesis, and the other (SC240) by adventitious regeneration). The complete resistance in the induced mutant was stable over two generations and was associated with a high frequency of leaf flecking, and consequently a low grain yield. Resistance in SC240 proved to be unstable; SC240 exhibited complete resistance to powdery mildew in the SC₂ and SC₃ generations, but only 20% of the SC₄ plants were completely resistant, while the remainder were indistinguishable in mildew response to ‘Guardian’. The mildew response of all the SC₅ generation of SC240 was not significantly different from ‘Guardian’. Yield analysis of the thirteen mutants with increased resistance in the presence of powdery mildew indicated that eleven exhibitedgrain yields at least as high as that of ‘Guardian’, while the mutant M19 exhibited a yield significantly higher than that of ‘Guardian’. This revised version was published online in July 2006 with corrections to the Cover Date.     

The oat line Pc54 as a source of resistance to crown rust,stem rust and powdery mildew in Europe

Summary The oat line Pc54 was found to be resistant to powdery mildew under both field and glasshouse conditions. The ratio of resistant to susceptible F₂ and F₂ progeny of a cross between a selection from the Pc54 line (Cc7422) and a susceptible cultivar (Selma) showed that, in addition to carrying the crown rust resistance gene Pc54 and the pg15 gene for stem rust resistance, the mildew resistance of the Pc54 line was conditioned by a single incompletely dominant gene along with additional factors which modified the expression of resistance. Previous results, that there was no linkage between genes Pc54 and Pg15, were confirmed. In addition, there was no evidence of linkage between the mildew resistance gene and gene Pc54. Evaluation of selections from within the Pc54 line showed that the expression of both stem rust and mildew resistance was modified by, or linked to, plant height. The effectiveness of genes Pc54 and Pg15, as measured by virulence frequencies, in central and eastern Europe is described.     

Conservation of powdery mildew resistance genes in three composite cross populations of barley

Summary Barley powdery mildew was used as a model to evaluate the potential of barley composite cross populations for conservation of disease resistance. The objective was to determine if increases in resistance to powdery mildew could be detected over periods of time in composite cross populations developed in California, where the disease might have had a selective influence on the populations, and the same populations grown in Montana, where no selective influence of powdery mildew was expected. Four isolates of Erysiphe graminis f. sp. hordei were used to monitor the frequencies of plants with specific mildew resistances through early, intermediate and late generations of three composite cross populations (CCII, CCV, CCXII) grown at Davis, California, and Bozeman and Moccasin, Montana. Changes in frequencies of plants resistant to the four isolates were observed between generations in all populations from the three locations. Trends in the frequencies of resistance are discussed in relation to selection pressure applied by E. graminis. It is suggested that associations with gene complexes other than resistance to E. graminis might help to explain the increased resistance observed in these studies.This research was funded in part by U.S. Agency for International Development Contract No. AID/DSAN-C-0024. The authors are grateful to Dr A. L. Kahler for seed of the composite cross populations and to Dr J. G. Moseman for the powdery mildew cultures.Cooperative investigations of the Agricultural Research Service, U.S. Department of Agriculture; the Department of Plant Pathology, Montana State University; and The Montana Agricultural Experiment Station. Journal Series Paper No. 1381 of the Montana Agricultural Experiment Station.     

Identification of Resistance Genes Against Powdery Mildew in Four Accessions of Hordeum Vulgare SSP. Spontaneum

Summary Four newly detected accessions of wild barley (Hordeum vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F₂ populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes.     

Genetic analysis of adult plant resistance to powdery mildew in pea (Pisum sativum L.)

Summary An analysis of adult plant resistance of powdery mildew in 15 F₁, F₂ and F₃ populations of pea derived from crossing 15 diverse and susceptible lines with one resistant line revealed that resistance to powdery mildew is controlled by duplicate recessive genes. The genes were designated as er₁ and er₂.Disease reaction showed independent segregation with three known markers in the resistant parent, namely, af (afila, chromosome 1), st (stipule reduced, chromosome 3) and tl (clavicula, chromosome 7).Contribution form the Department of Genetics and Plant Breeding Banaras Hindu University, Varanasi-221005, India.     

Hordeum chilense resistance to powdery mildew and its potential use in cereal breeding

Summary Hordeum chilense is a wild barley with high crossability with Triticum, Hordeum and Secale. Its amphiploid with wheat, tritordeum, has potential as a new crop. H. chilense is highly resistant to the powdery mildew diseases of both wheat and barley. Whereas tritordeum is resistant to barley powdery mildew, its reaction to wheat powdery mildew is similar to that of its wheat parent. However H. chilense contributes to a reduced density of mildew colonies. This quantitative resistance of tritordeum is diluted at higher ploidy levels.     

Inheritance of the Powdery Mildew Resistance Mlk in Wheat

The inheritance of the Mlk powdery mildew resistance originating from ‘Heine 2174.50’ was analyzed by crossing the Mlk resistant cultivar ‘Ralle’× cv. ‘Amor’ (highly susceptible) and vice versa and by observing the reactions of F₁- and F₂-plants after inoculation with two different Mlk avirulent powdery mildew isolates. In all cases, a 3 (resistant): I (susceptible) segregation was found in F₂. The reactions of the F₂plants against the two powdery mildew isolates were identical in each case. Therefore, it is supposed that one dominant resistant gene is responsible for the resistant reactions against these two isolates. These results support the earlier assumption of Heun and Fischbeck (1987b) that the whole Mlk resistance pattern is controlled by a single gene.     

Spectrum of resistance conferred by ML-O powdery mildew resistance genes in barley

Summary Ten barley mutants and five Ethiopian barley lines representing 11 independently arisen powdery mildew resistance genes in the ml-o locus were tested at the seedling stage to cultures of the powdery mildew fungus from Europe, Israel, USA. Canada, and Japan. They were resistant with infection type 0/(4) in all tests. They were also resistant to field populations of the pathogen when scored in disease nurseries at more than 78 locations in 29 countries in Europe, the Near East, North and South America. New Zealand, and Japan. This indicates that the 11 genes confer the same, world-wide spectrum of powdery mildew resistance. They have no effect on several other barley diseases such as stripe rust and leaf rust.Part of the research reported here was carried out under IAEA Research Agreement No 1043 and Research Contract No 139-74-1 BIO DK with the European Atomic Energy Community.     

Isolation, chromosomal location, and expression analysis of putative powdery mildew resistance genes in wheat (Triticum aestivum L.)

Special and degenerate primers are designed according to the conservative sequence of barley powdery mildew resistance genes Mla1, Mla6, and Mla13. Two wheat Mla-like orthologs, TaMla-2 and TaMla-3 are cloned and sequenced from the cDNA of wheat resistant-powdery mildew line TAM104R by RT-PCR method. TaMla-2 and TaMla-3 encode distinct but highly related coiled-coil nucleotide-binding site leucine-rich repeat type (NBS-LRR) resistant disease proteins and both reveal about 74 and 81% identity with amino acid sequence of Mla1, respectively. They are multiple copies in wheat genomes, one copy of them is mapped on wheat chromosome 1AL and two on 1BL using Chinese Spring nulli-tetra-somic lines and ditelosomic lines of 1A, 1B and 1D in southern analysis. This result suggests that may be the two Mla-like genes originated from the two diploid ancestral genomes, respectively. The expression pattern analysis of semi-quantitative PCR shows the TaMla genes are mainly expressed in leaf and sheath, and expression level is enhanced in organs infected by Erysiphe graminis, suggesting that TaMla-2 and TaMla-3 are powdery mildew resistance related-genes in wheat. Electronic Supplementary Material Supplementary material is available for this article at     

The resistance of Hordeum bulbosum and its hybrids with H. vulgare to common fungal pathogens

Summary Two tetraploid and two diploid clones of Hordeum bulbosum were screened for resistance to five isolates of powdery mildew which are virulent on cultivated barley. All were resistant and this resistance was also expressed in hybrids with H. vulgare. The tetraploid genotypes were also resistant to isolates of yellow rust and brown rust. These results show that H. bulbosum contains useful genes for resistance to these diseases and that there is a potential to transfer these into cultivated barley.     

Single-gene resistance to powdery mildew in zucchini squash (Cucurbita pepo)

Cucurbita pepo (pumpkin, squash,gourd) is an economically important species that is susceptible to the cucurbit powdery mildew fungus, Podosphaera xanthii(syn. Sphaerotheca fuliginea). ‘True French’, an open-pollinated cultivar of the Zucchini Group of C. pepo, was crossed with an unnamed powdery-mildew resistant straight neck-type accession, the resistance of which was apparently derived from an interspecific cross with a resistant wild species of Cucurbita,and resistant plants were selected in the F₂ generation. This was followed by six cycles of backcross-pedigree selection for resistance, and resulted in the development of an accession true-breeding for resistance to powdery mildew and nearly isogenic to ‘True French’. The resistant and susceptible near-isogenics were crossed and seeds of the filial and backcross generations were produced. Plants of the parental accessions and their progenies were grown together in a controlled-environment chamber, exposed to the pathogenic fungus, and scored as resistant, partially resistant, or susceptible 27–33 days after sowing. The results indicated that resistance is conferred by a single incompletely dominant gene, designated Pm-0. This revised version was published online in August 2006 with corrections to the Cover Date.     

Screening of melon (Cucumis melo L.) germplasm for multiple disease resistance

Summary Melon germplasm was screened for cucumber green mottle mosaic virus (CGMMV), powdery mildew (Sphaerotheca fuliginea), downy mildew (Pseudoperonospora cubensis) and Fusarium wilt (Fusarium oxysporum f. sp. melonis) resistance under artificial conditions except downy mildew for which screening was done under natural epiphytotic conditions. High level resistance to all the four diseases was not recorded in any of the collections tested. Nevertheless, ertheless, resistance to three diseases was located in three germplasm. Wild Cucumis species C. figarei exhibited absolute resistance to CGMMV and Fusarium wilt and high level resistance to downy mildew. Phoot or snapmelon (Cucumis melo var. momordica) — a non-dessert from of Indian origin—was highly resistant to downy mildew and resistant to CGMMV and medium resistant to Fusarium wilt. Iroquois was resistant to powdery mildew and medium resistant to downy mildew and CGMMV.     

Molecular characterization of a novel powdery mildew resistance gene Pm30 in wheat originating from wild emmer

Powdery mildew caused by Erysiphe graminis f. sp. tritici is one of the most important wheat diseases in many regions of theworld. A powdery mildew resistance gene, originating from wild emmerwheat (Triticum dicoccoides) accession `C20', from Rosh Pinna, Israel,was successfully transferred to hexaploid wheat through crossing andbackcrossing. Genetic analysis indicated that a single dominant genecontrols the powdery mildew resistance at the seedling stage. SegregatingBC₁F₂ progenies of the cross 87-1/C20//2*8866 wereused for bulked segregant analysis (BSA). The PCR approach was used togenerate polymorphic DNA fragments between the resistant and susceptibleDNA pools by use of 10-mer random primers, STS primers, and wheatmicrosatellite primers. Three markers, Xgwm159/430,Xgwm159/460, and Xgwm159/500, were found to be linked tothe resistance gene. After evaluating the polymorphic markers in twosegregating populations, the distance between the markers and the mildewresistance gene was estimated to be 5–6 cM. By means of ChineseSpring nullisomic-tetrasomics and ditelosomics, the polymorphic markersand the resistance gene were assigned to chromosome arm 5BS and werephysically mapped on the gene rich regions of fragment length (FL) 0.41–0.43 by Chinese Spring deletion lines. As no powdery mildew resistancegene has been reported on chromosome arm 5BS, the mildew resistancegene originating from C20 should be a new gene and is designated Pm30.     

Genetic linkage of the Yr1 and Pm4 genes for stripe rust and powdery mildew resistances in wheat

Summary Genes Yr1 for resistance to stripe rust and Pm4a for resistance to powdery mildew showed linkage of 2.0±0.6 cM. Close repulsion linkage probably accounts for the absence in European wheats of genes Yr1 and Pm4b in combination.     

Shading of plants facilitates selection for powdery mildew resistance in Squash

Summary Powdery mildew development was assessed on squash (Cucurbita pepo) plants of a susceptible cultivar, a resistant accession, their F₁, and their F₂ in an early summer planting in the field, covered or not covered with a shading net. Three reaction types were observed: susceptible, powdery mildew on stems and on both, the upper and lower leaf surfaces, as in the susceptible parent; resistant, no powdery mildew on leaves or stems, as in the resistant parent; and partially resistant, powdery mildew on upper leaf surfaces only, as in the F₁. Disease presence on the stem was associated with susceptibility. Shading hastened the appearance of powdery mildew and increased the severity of infection on partially resistant and susceptible plants, facilitating identification of resistant individuals in the F₂ population.Contribution No. 1613-E from the Agricultural Research Organization, Bet Dagan, Israel