Identification of growth stage dependent expression of partial resistance of barley to powdery mildew

Summary Genotypic differences for growth stage dependent expression of partial resistance to barley powdery mildew have been identified on the basis of two components of resistance in the glasshouse and the pattern of epidemic development in the field.Differences for infection frequency and proportion of sporulating colonies were highly significant between the 16 genotypes investigated at four stages of plant development in the glasshouse. Both resistance components were significantly correlated mutually (r=0.73 to 0.86) and with the infection level on leaves developed at comparable growth stages in the field (r=0.52 to 0.73). The infection level of seedlings in the glasshouse was significantly correlated with the infection level of the leaves of the first until the third node in the field (r=0.70 to 0.73). Adult plant resistance was predominantly expressed at the uppermost leaf. A clear genotype × growth stage interaction was apparent for infection level; some genotypes showed partial resistance predominantly at the seedling stage and others predominantly at the adult plant stage.Abbreviations IF Infection Frequency, number of colonies per cm² leaf area, glasshouse experiment - IL Infection Level, number of colonies per cm² leaf area, field experiment - PSC Proportion Sporulating Colonies from total number of colonies, glasshouse experiment     

Diallel analysis of partial resistance to powdery mildew caused by Erysiphe graminis f. sp. hordei in spring barley (Hordeum vulgare L.)

Summary Six spring barley genotypes with growth stage dependent expression of partial resistance to powdery mildew were crossed following a half diallel scheme. Data for percentage infected leaf area at different growth stages from the F1 and F2 generation and the F2 offspring were analyzed. The absence of a contrast between parents and offspring indicated that dominance effects were not important at the young growth stages. Diallel analysis showed that general combining ability effects (GCA) were important at all growth stages. Although significant, specific combining ability effects were of minor importance. The cross between the most resistant parents with the largest, negative GCA resulted in the most resistant progeny.     

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.     

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.     

Inheritance of resistance to powdery mildew (Erysiphe graminis f. sp. hordei) in eleven primitive barley varieties

The inheritance of the powdery mildew resistance of eleven primitive barley varieties was investigated. crosses with varieties with identified resistance genes revealed that at least three different genes are present in a group of six genotypes from Greece (Hor736, Hor847, Hor878, Hor1159, Hor1379 and Hor1873) and one from Turkey (Hor1188). One dominant gene is common to these seve genotypes and in the Greek accessions a second gene is present. The expression of this second gene was respectively dominant in the crosses with Hor736 and Hor1159, recessive in the crosses with Hor878 and Hor1873, and depended on the employed powdery mildew isolate in the crosses with Hor847 and Hor1379. The detected genes differ from the resistance genes of the crossing partners, viz. Ml-al2, Ml-(Ab), Ml-(CP) and Ml-(1402). The accessions from China (Hor824 and Hor4021) each possess two resistance genes which differ from Ml-a12, Ml-(La) and Ml-(CP). The major gene is common to both accessions and is either identical, allelic or closely linked to a gene in the variety Nigrate. The Columbian accession Hor1894 possesses one resistance gene linked to a resistance gene in Nigrate but this gene differs from the Ml-a locus. The variety Palestine (Hor3997) possesses two resistance genes of which one is allelic or closely linked to Ml-(at).     

Differences in partial resistance of barley to powdery mildew (Erysiphe graminis DC F.SP. hordei Marchal) after chemomutagenesis. II. Reaction of mutants to pathotypes

Summary Seedlings of 45 barley mutants with partial resistance in the field and their parental cultivars Asse, Bomi, and Vada were exposed to six local and two foreign races of powdery mildew, in climatically controlled cabinets. Infectability, pustule size, infection grade, and infection type were estimated. No mutant did demonstrate strong race-specific reactions although some race-specific interactions of moderate grade could be detected. The results of infection of mutants with single races confirmed the quantitative character of their change in resistance determined in the earlier field assessments. The 20 investigated mutants with increased resistance expressed a lower level of disease with each of the used races. Out of the 10 mutants with higher susceptibility in the field, 4 mutants again exhibited higher degrees of infection over all the different races, while the remaining 6 mutants were not distinguishable from the original cultivar under the given growing conditions. Out of the tested 14 developmentally resistant mutants, only the 5 genotypes with different seedling reaction could be analyzed for race-specificity in this study. In all the above cases as the result of one gene mutation, quantitative shifts in level of infection were recorded over all the 8 races.Qualitative estimates of infection type were supported by data on the frequency of chlorotic or necrotic lesions and green islands on the infected leaves. But quantitative methods of assessment, e.g. infection frequency and pustule size, were more effective in studying partial resistance of the mutants. Infection grade, estimated visually 14 days after inoculation, was in agreement with the quantitative parameters accurately measured 7 days after inoculation. Therefore, its careful use can be recommended to speed the screening by narrowing down the materials. Differences in virulence level of the races were observed and their influence on race-specificity studies was discussed.     

Evaluation of sources of partial resistance to mildew in barley using enzyme-linked immunosorbent assay and other assessment methods

Summary Twenty-nine spring barley genotypes ranging from primitive landraces to modern cultivars were characterised for partial resistance to powdery mildew on detached leaves and in the field. Detached seedling and adult plant leaves were produced under spore-free glasshouse conditions and were assessed for infection frequency (IF) and then total mildew biomass using enzyme-linked immunosorbent assay (ELISA), and thereby biomass per colony (BMC), following controlled infection. Field plants were assessed visually for infection and for total mildew biomass using ELISA. Several genotypes were identified as having below average IF and BMC on both seedling and adult detached leaves and field scores of leaf area infected with mildew. IF at the seedling stage was highly correlated with IF at adult plant stage and with mildew biomass in the field. Adult plant IF was highly correlated with visual field scores. BMC data were often negatively correlated with other assessments.     

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.     

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.     

Detection of tolerance of barley cultivars to infection by powdery mildew (Erysiphe graminis f.sp. hordei)

Summary Barley genotypes representing a wide range of resistance expressions and origins, from major resistance genes in modern cultivars to field resistances in landraces, were assessed for tolerance to disease under glasshouse and field conditions. A few genotypes were picked out as showing less yield loss than would be expected from the level of mildew infection. Genotypes showing more than the expected yield loss were also found. The potential use of tolerance as a breeding character is discussed.     

Partial resistance to Erysiphe graminis hordei in old European barley varieties

Summary A collection of 133 genotypes consisting mainly of old European barleys and land varieties was assessed for level of partial resistance to barley mildew at the adult plant stage. Identification of any known resistance factors conferring hypersensitive reactions to the mildew races present at various frequencies in N.W. Europe was ensured through seedling tests with an appropriate AO mildew isolate.Thirty-nine varieties with relatively high levels of adult (partial) resistance and lacking known race-specific resistance genes were tested over a period of three years in field disease nurseries. At least thirteen varieties showed significantly less percentage leaf area infected than the control variety Proctor. These constitute a useful reserve of non-hypersensitive resistance, probably under the control of genes with mainly additive effects. In combination, these genes give enhanced levels of resistance, which could be of a more stable nature.     

Associations between three ml-o powdery mildew resistance genes and agronomic traits in barley

Summary A population of 198 chromosome-doubled haploid lines of spring barley was scored for segregation in locus ml-o (powdery mildew reaction) on chromosome 4 and in the linked loci s (rachilla hair length) and ddt (reaction to the insecticide DDT) on chromosome 7. They were also tested in a disease-free field trial for the agronomic traits: grain yield, thousand grain weight, lodging, and necrotic leaf spotting. The three mutagen-induced resistance genes ml-o5, ml-o6 (from Carlsberg II) and ml-10 (from Foma) showed no detectable differences with respect to effects on agronomic traits. They all conferred a four per cent reduction in grain yield caused mainly by lower thousand grain weight, and an increase in necrotic leaf spotting. The two original mutants of Carlsberg II had additional mutant genes affecting agronomic traits. Lines with gene S (long hair) had on average a three per cent higher thousand grain weight than those with s. The alleles in locus ddt showed no association with the agronomic traits. It is concluded i) that the associations between the three ml-o alleles and agronomic traits are caused by pleiotropy, ii) that ml-o resistant, high-yielding lines may be selected, and iii) that the association between gene s and thousand grain weight may be due to genetic linkage.Abbreviations DH-lines chromosome-doubled haploid lines     

New molecular markers linked to qualitative and quantitative powdery mildew and scald resistance genes in barley for dry areas

A partial genetic linkage map was constructed on 71 doubled-haploid lines derived from a cross between the barley lines Tadmor and WI2291 with 181 molecular markers. The segregating population was used to detect markers linked to the gene Mlg conferring resistance to powdery mildew (Erysiphe graminis f. sp. hordei) and to genes for quantitative resistance to scald (Rhynchosporium secalis). The gene Mlg on chromosome 4H was flanked by two AFLP markers at a distance of 2.0 and 2.4 cM, respectively. QTLs for resistance to scald were detected on chromosomes 2H and 3H. This association of molecular markers with qualitative and quantitative disease resistance loci represents a valuable starting-point for marker-assisted selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Characterisation of new oat germplasm for resistance to powdery mildew

Summary Wild oat accessions from the Institute of Plant Genetics and Crop Plant Research (IPK) Gatersleben were selected as potential sources of resistance to powdery mildew (Erysiphe graminis f.sp. avenae). The Avena strigosa accessions AVE 128, AVE 488 and AVE 264, along with A. occidentalis accession CAV 3889 showed high levels of mildew resistance in tests at early and late growth stages over two years; the latter is a particularly useful source because it is a hexaploid species. Moderate levels of resistance were also found in A. fatua accession AVE 1981, AVE 2032 and A.sterilis accession AVE 1373 but this could be due to the later maturity of these genotypes.     

Barley mildew in Europe: population biology and host resistance

Summary Isolates of the barley mildew pathogen from the air spora over a large part of Europe and from fields of variety mixtures, were tested for virulence against 12 host resistance alleles. Subsamples were tested for their response to triadimenol fungicide and analyzed for 10 DNA loci using RAPD markers and PCR. There was a large range of haplotypes spread over Europe; irregularity in the distribution was probably due mainly to non-uniform use of the corresponding host resistances and fungicides. A large range of variation was also detectable within individual fields. Positive gametic disequilibria distorted the distribution of virulence alleles among haplotypes and reduced the number of haplotypes detectable in the sample. Analysis of the spread of the newly selectedVal3 allele into different European sub-populations indicated that gene flow throughout the population may be rapid for alleles that have a selective advantage.Fungicide resistance was widespread in areas known for intensive use of fungicides for mildew control. Four classes of fungicide response were detectable and particular virulence haplotypes were found to be characteristic for each class.Variety mixtures used in the former German Democratic Republic (GDR) reduced mildew infection, and thus fungicide use, during the years 1984–1991 despite the limited variation in host resistance among the mixtures. A tendency for complex pathogen races to increase in mixture crops was reversed by the large-scale re-introduction of fungicides for mildew control in 1991. The mixture strategy appeared to be more successful than using the same resistance alleles in pure monoculture or combining them in a single host genotype.     

Two unnecessary powdery mildew resistance genes in a synthetic rye population are neutral on fitness

A synthetic winter rye population was produced with two race-specific powdery mildew resistance genes, one dominant (Rm1) and the other (rm2) recessive, each at a frequency of about 0.50. The population was advanced by open-pollination in an isolated plot under mildew-free conditions for eight years. Samples of generations Syn-0 through Syn-7 were inoculated in the laboratory with two mildew isolates, one avirulent to either resistance gene, the other virulent to Rm1 and avirulent to rm2, to discriminate resistant and susceptible phenotypes. From the proportions of resistant plants, frequencies of Rm1 and rm2 were calculated and the fitness of carriers of resistance alleles was estimated in relation to carriers of susceptibility alleles at the two loci using continuous models and linear regression analyses. Frequencies of the two resistance genes oscillated only weakly over the eight generations. Coefficients of selection against Rm1-and rm2rm2 genotypes were –0.04 and –0.02, respectively, and not significantly different from zero. Thus the two resistance genes were selectively neutral. It is concluded that pyramiding of major powdery mildew resistance genes in rye varieties should not reduce their yield potential in the absence of mildew.     

Involvement of polyamines, diamine oxidase and polyamine oxidase in resistance of barley to Blumeria graminis f. sp. hordei

Polyamine levels and diamine and polyamine oxidase activities have been investigated in the first leaves of barley (Hordeum vulgare L.) in the absence of or following inoculation with conidia of powdery mildew (Blumeria graminis f. sp. hordei). Two cultivars with varying sensitivity to powdery mildew, viz., Chariot (resistant) and Golden Promise (susceptible) were used. The levels of putrescine, spermidine and spermine were found to be higher in the leaves of Chariot than in the leaves of Golden Promise and, with the exception of spermine, were generally higher in both cultivars after inoculation. In inoculated leaves of Chariot, levels of putrescine and spermidine peaked at 9 days and 12 days, respectively. In controls (uninoculated leaves), the activities of these enzymes, and putrescine and spermidine levels also increased but not to the same extent as in inoculated leaves. With Golden Promise, the levels of putrescine and spermidine in the inoculated leaves changed very little over sampling times but were always higher than in the controls. In this cultivar, there was little difference between inoculated leaves and the controls in diamine oxidase activity which reached a maximum value at 9 days post-inoculation. Activity of the bound form of diamine oxidase was low in both the cultivars. Polyamine oxidase was not detected at 3 days after inoculation in either cultivar but activity at fairly low levels was recorded at later times, usually reaching a maximum value at 9 days. The results suggest that polyamine metabolism and diamine oxidase activity in particular may be involved in the mechanism conferring resistance to barley powdery mildew in Chariot.     

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.     

Resistance to powdery mildew in a doubled haploid barley population and its association with marker loci

The genetic basis of resistance to powdery mildew (Erysiphe graminis DC. f.sp. hordei Marchal) was analyzed using doubled haploid barley (Hordeum vulgare L.) lines from the cross Harrington/TR306. Based on infection types observed after inoculation with defined single-conidium isolates, the lines were classified into four groups. The observed phenotypic ratio fit a two-locus model. The two putative loci were mapped relative to molecular markers. One coincided with the previously mapped dMlg locus on chromosome 4. Based on the observed infection types, Harrington carries the Mlg resistance allele, and TR306 carries a second locus on chromosome 7 (5H); this was tentatively designated Ml(TR). It is the first reported race-specific powdery mildew resistance gene located on that chromosome. These two loci were also detected by simple interval mapping of disease severity data from naturally infected field plots. Composite interval mapping with the first two resistance loci as co-factors detected an additional locus on chromosome 6, with a minor effect on resistance. Finally, superimposing the race-specific classification onto the field data provided evidence for a minor-effect locus on chromosome 7 (5H). The Mlg locus had the largest effect, the Ml(TR) locus had an intermediate effect and the other two loci had very small effects. This study demonstrates the effectiveness of an integrated approach to identifying and mapping resistance loci using classification data from inoculated experiments and quantitative data from field experiments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Regional distribution of resistances to powdery mildew in winter and spring barley cultivars grown in the northern part of France

The spatio-temporal distribution of race-specific resistances to powdery mildew was analysed in northern France (the east, the north and the west of Paris). Resistances were identified in 26 winter and six spring barley cultivars. Seedling leaf segments were inoculated with 20 powdery mildew isolates, chosen to identify 14 resistance alleles. As opposed to other European countries, the resistance alleles differed between winter and spring cultivars grown in the three regions. Most of the winter cultivars had no resistance allele, or only the widespread resistance alleles Mlra and/or Mlh, plus Mlg in the west. Mla9 and Mla13 were also present in the north, but at a low frequency. Spring cultivars carried the alleles Mla7, Mla9, Mla12, Mlk, Mlg or MlLa in the east, where a diversification of resistances has occurred since 1987, particularly because of the use of ‘Volga’ (Mla7, Mlk, Mlg and MlLa). In the north and the west, Mla12 dominated after a decrease in the frequency of Mla7, Mla13 has recently been introduced in the north and the west with the cultivar ‘Vodka’.