Temporary partial breakdown of mlo-resistance in spring barley by sudden relief of soil water-stress under field conditions: the effects of genetic background and mlo allele

This study, carried out under field conditions, assessed the extent to which temporary breakdown of mlo- resistance, following relief of water-stress, was determined by genetic background and mlo allele. Commercial barley cultivars expressing the mlo gene for resistance to powdery mildew ( Erysiphe graminis ( Blumeria graminis ) f.sp. hordei ) were tested as well as doubled haploid progeny from spring barley genotypes, a proportion of which were sown in the field in two successive years. Plants were protected from natural rain by a mobile rain shelter and either watered by trickle-irrigation or allowed to dry. Percentage mildew infection resulting from natural inoculum was recorded and the doubled haploid genotypes were classified as resistant, intermediate or susceptible on the basis of their control (watered) treatment scores. In each of the three designated classes, particular genotypes developed infection levels, following relief of water-stress, that were higher than those observed on the well-watered controls. This was found not to be related to the mlo allele as there was no significant difference between the increases observed on resistant plants carrying mlo9 and resistant plants carrying mlo11 . Differences in the degree of breakdown of resistance were attributed to genetic background rather than to the specific mlo allele.     

Resistance, epidemiology and sustainable management of Rhynchosporium secalis populations on barley

Rhynchosporium secalis is one of the most destructive pathogens of barley worldwide, causing yield decreases of up to 40% and reduced grain quality. Rhynchosporium is a polycyclic disease. Primary inoculum includes conidia produced on crop debris, infected seeds and possibly ascospores, although these have not yet been identified. Secondary disease spread is primarily by splash dispersal of conidia produced on infected leaves, which may be symptomless early in the growing season. Host resistance to R. secalis is mediated by both 'major' or host-specific genes (complete resistance) and 'minor' genes of smaller, generally additive effects (partial resistance). Crop growth stage and plant or canopy architecture can modify the expression of resistance. Resistance genes are distributed unevenly across the barley genome, with most being clustered on the short arms of chromosomes 1H, 3H, 6H and 7H, or in the centromeric region or on the long arm of chromosome 3H. Strategies used to manage rhynchosporium epidemics include cultivar resistance and fungicides, and also cultural practices such as crop rotation, cultivar mixtures and manipulation of sowing date, sowing rate or fertiliser rate. However, the high genetic variability of R. secalis can result in rapid adaptation of pathogen populations to render some of these control strategies ineffective when they are used alone. Sustainable control of rhynchosporium needs to integrate major-gene-mediated resistance, partial resistance and other strategies such as customized fungicide programmes, species or cultivar rotation, resistance gene deployment, clean seed and cultivar mixtures.     

Interaction between powdery mildew and barley with mlo5 mildew resistance

Powdery mildew infection of barley with the mlo5 barley powdery mildew resistance gene was examined, using near-isogenic barley lines, with and without mlo5 resistance, and two near-isogenic powdery mildew isolates, HL3/5 and GE3 with high (virulent) or low (avirulent) penetration efficiency on the resistant barley line. In all isolate–host combinations (except GE3 on the resistant barley line), frequency of haustorium formation increased significantly from zero at 11 h after inoculation to a maximum by 13 h, and there was no subsequent increase up to 24 h. In the susceptible barley line, 27% of appressoria from both isolates formed haustoria. Although this was significantly higher than the frequency of haustorium formation (18%) of HL3/5 on the resistant barley line, HL3/5 was much more successful than GE3 (frequency of haustorium formation less than 1%). The fact that HL3/5 did not possess a generally higher ability to penetrate successfully to form haustoria on the susceptible barley line, indicates that HL3/5 did not overcome the mlo5 resistance by being generally more vigorous. In the resistant barley line, papillae were larger than in the susceptible line; however, both isolates were associated with papillae of the same diameter at the time of penetration. We suggest that the mlo5 resistant barley line confers two different forms of resistance: isolate-specific and isolate-nonspecific.     

Application of real-time and multiplex polymerase chain reaction assays to study leaf blotch epidemics in barley

ABSTRACT Leaf blotch, caused by Rhynchosporium secalis, was studied in a range of winter barley cultivars using a combination of traditional plant pathological techniques and newly developed multiplex and real-time polymerase chain reaction (PCR) assays. Using PCR, symptomless leaf blotch colonization was shown to occur throughout the growing season in the resistant winter barley cv. Leonie. The dynamics of colonization throughout the growing season were similar in both Leonie and Vertige, a susceptible cultivar. However, pathogen DNA levels were approximately 10-fold higher in the susceptible cultivar, which expressed symptoms throughout the growing season. Visual assessments and PCR also were used to determine levels of R. secalis colonization and infection in samples from a field experiment used to test a range of winter barley cultivars with different levels of leaf blotch resistance. The correlation between the PCR and visual assessment data was better at higher infection levels (R(2) = 0.81 for leaf samples with >0.3% disease). Although resistance ratings did not correlate well with levels of disease for all cultivars tested, low levels of infection were observed in the cultivar with the highest resistance rating and high levels of infection in the cultivar with the lowest resistance rating.     

Characterization of DMI,QoI and SDHI fungicides sensitivity of Ramularia collo-cygni isolates in Argentina

Journal of Plant Diseases and Protection - Ramularia leaf spot (RLS) caused by the fungus Ramularia collo-cygni (Rcc) has become a threat to barley production in Argentina. All barley varieties are...     

A hypothesis on physiological alterations during plant ontogenesis governing susceptibility of winter barley to ramularia leaf spot

The occurrence and epidemic spread of ramularia leaf spot (RLS) caused by Ramularia collo-cygni was studied with relation to the plant age-dependent alterations in the antioxidative defence systems in the leaves during mature stages of field-grown winter barley. The breakdown of enzymatic activities of dehydroascorbate reductase, monodehydroascorbate reductase and glutathione reductase in the two uppermost fully expanded leaf layers correlated well with a decrease in the pool of the non-enzymatic antioxidants ascorbate and glutathione. The general decline in the antioxidative systems occurred between 31 May and 8 June after ear emergence stages and preceded the first visible symptoms of RLS. The activities of peroxidases corresponded to an increase in phenolics and lignification which seemed to enhance the infectivity of the fungus rather than to protect the plant. The first R. collo-cygni conidia were trapped in the field about 4 weeks before the latent infection was first detected by ELISA, and 5 weeks before the first disease symptoms became visible. Both the counts of airborne conidia and the weather data recorded between April and June suggest that neither inoculum nor the weather conditions are limiting factors for infection by R. collo-cygni at earlier growth stages of barley. It is concluded that the relatively late development of RLS in the field in mid June is governed by the significant degradation of the antioxidative protection systems in the leaves of barley at the onset of ripening stages, rather than by environmental factors or the availability of airborne inoculum.     

Genetic Structure of Rhynchosporium secalis in Australia

ABSTRACT Restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of Australian field populations of the barley scald pathogen Rhynchosporium secalis. Fungal isolates were collected by hierarchical sampling from five naturally infected barley fields in different geographic locations during a single growing season. Genetic variation was high in Australian R. secalis populations. Among the 265 fungal isolates analyzed, 214 distinct genotypes were identified. Average genotype diversity within a field population was 65% of its theoretical maximum. Nei's average gene diversity across seven RFLP loci was 0.54. The majority (76%) of gene diversity was distributed within sampling site areas measuring 1 m(2); 19% of gene diversity was distributed among sampling sites within fields; and 5% of gene diversity was distributed among fields. Fungal populations from different locations differed significantly both in allele frequencies and genotype diversities. The degree of genetic differentiation was significantly correlated with geographic distance between populations. Our results suggest that the R. secalis population in Western Australia has a different genetic structure than populations in Victoria and South Australia.     

Virulence spectrum to barley (Hordeum vulgare L.) in isolates of Rhynchosporium secalis from Syria

Journal of Plant Diseases and Protection - Scald caused by Rhynchosporium secalis, is an economically important disease found worldwide. Information on the variability of virulence in R. secalis is...     

A Japanese powdery mildew isolate with exceptionally large infection efficiency on Mlo-resistant barley

A Japanese field isolate (Race I) of Erysiphe graminis f.sp, hordei was tested on 17 barley lines carrying the mlo powdery mildew resistance gene. Race I produced many successful infections with infection type larger than or equal to 2 on six lines (M66, MC20, SRI, SR7, A tem and Totem), On the remaining 11 lines it reacted with infection type 0. Colony numbers on the Mlo-lines were between 26% and 12 9% of the numbers on a susceptible cultivar Manchuria, These numbers were larger than, or similar to those produced by isolate HL3/5, which has the highest recorded infection efficiency on Mlo-resistant cultivars. The interaction between isolates and lines was highly significant. The isolate GE3, from which HL3/5 was derived by selection, gave rise to occasional colonies corresponding to less than 03° o of the number produced on cv. Manchuria.     

Virulence spectrum in populations of the barley powdery mildew pathogen, Erysiphe graminis f. sp. hordei in Tunisia and Morocco in 1992

The virulence of 57 single colonies of Erysiphe graminis f. sp. hordei from Tunisia and Morocco was investigated using the Pallas near-isogenic differential set. Virulence patterns in general were similar in both countries. Va8, Va10+Du2, V41/145, VLa and VRu2 appear to be common in the region. The resistance alleles Mla7 and Mla9, in combination with other resistances, were highly effective against the isolates tested. No virulence against mlo and Mla9 + k was detected in Tunisia. No virulence on Mla7 and Mla9 was detected in Morocco. The frequency of virulence against several resistances was significantly higher in Morocco than in Tunisia. On the other hand, virulence against other resistances was higher in Tunisia than in Morocco. Three isolates from Morocco infected mlo to an extent greater than previously described. All pathotypes were unique. An attempt is made to interpret the results by comparison with pathotype evolution in Europe.     

Differential Selection on Rhynchosporium secalis During Parasitic and Saprophytic Phases in the Barley Scald Disease Cycle

ABSTRACT Competition among eight Rhynchosporium secalis isolates was assessed during parasitic and saprophytic phases of the disease cycle in field experiments conducted at two locations and over two growing seasons. The eight isolates were inoculated onto six barley populations exhibiting varying degrees of resistance. Microsatellite analysis of 2,866 isolates recovered from the field experiments showed significant, and sometimes opposite, changes in the frequencies of R. secalis genotypes during the growing season (parasitic phase) and between growing seasons (saprophytic phase). Isolates that showed the most complex virulence in greenhouse seedling assays had the lowest fitness in the field experiment. Significant differences in isolate fitness were found on different host populations and in different environments. Selection coefficients were large, indicating that evolution can occur rapidly in field populations. Although inoculated isolates had the lowest overall fitness on the moderately resistant landrace cv. Arabi Aswad, some isolates were more virulent and consistently increased in frequency on this landrace, suggesting a risk of directional selection and possible erosion of the resistance following its widespread deployment in monoculture. These results provide the first direct evidence that R. secalis pathogen genotypes differ in their saprophytic ability and parasitic fitness under field conditions.     

The effect of component number on Rhynchosporium secalis infection and yield in mixtures of winter barley cultivars

Mixtures of winter barley cultivars containing up to six components were grown over three years with and without fungicide treatment. Yield increases were recorded for mixtures compared with the mean of their monoculture components and there was a significant trend towards greater benefit from increased number of components. These benefits were partially attributable to a corresponding increase in control of Rhynchosporium secalis as component number increased. The potential for exploitation of mixtures in cereals for control of splash-dispersed pathogens is discussed.     

A Compromised Mlo Pathway Affects the Response of Barley to the Necrotrophic Fungus Bipolaris sorokiniana (Teleomorph: Cochliobolus sativus) and Its Toxins

ABSTRACT In search of new durable disease resistance traits in barley to control leaf spot blotch disease caused by the necrotrophic fungus Bipolaris sorokiniana (teleomorph: Cochliobolus sativus), we developed macroscopic and microscopic scales to judge spot blotch disease development on barley. Infection of barley was associated with cell wall penetration and accumulation of hydrogen peroxide. The latter appeared to take place in cell wall swellings under fungal penetration attempts as well as during cell death provoked by the necrotrophic pathogen. Additionally, we tested the influence of a compromised Mlo pathway that confers broad resistance against powdery mildew fungus (Blumeria graminis f. sp. hordei). Powdery mildew-resistant genotypes with mutations at the Mlo locus (mlo genotypes) showed a higher sensitivity to infiltration of toxic culture filtrate of Bipolaris sorokiniana as compared with wild-type barley. Mutants defective in Ror, a gene required for mlo-specified powdery mildew resistance, were also more sensitive to Bipolaris sorokiniana toxins than wild-type barley but showed less symptoms than mlo5 parents. Fungal culture filtrates induced an H2O2 burst in all mutants, whereas wild-type (Mlo) barley was less sensitive. The results support the hypothesis that the barley Mlo gene product functions as a suppresser of cell death. Therefore, a compromised Mlo pathway is effective for control of biotrophic powdery mildew fungus but not for necrotrophic Bipolaris sorokiniana. We discuss the problem of finding resistance traits that are effective against both biotrophic and necrotrophic pathogens with emphasis on the role of the anti-oxidative system of plant cells.     

Effect of barley yellow dwarf virus on susceptibility of barley cultivars to net blotch (Pyrenophora teres) and leaf blotch (Rhynchosporium secalis)

The effect of barley yellow dwarf virus (BYDV) on the development of net blotch (Pyrenophora eves) and leaf blotch (Rhynchosporium secalis) was examined on seven barley cuitivars. Seedlings were infected with BYDV at the two-leaf stage (G.S. 12). Their susceptibility to three isolates of p teres and isolates of two races (U.K. 1, U.K. 2) of R. secalis was examined at the four-leaf stsge (G.S. 14) and when plants were more mature (G.S. 33/38). At G.S. 14 numbers of lesions produced by P. teres and R. secalis were reduced, on average, by 37 and 72% respectively, and at G.S. 33/38 by 61 and 74%.     

Quantification of seedborne infection by Rhynchosporium secalis in barley using competitive PCR

The potential of the competitive polymerase chain reaction (PCR) assay for quantification of seedborne infection by Rhynchosporium secalis in barley was examined using a primer set (RS1 and RS3) derived from the internal transcribed spacer (ITS) regions of ribosomal RNA genes of this pathogen. Introduction of a heterologous internal control, which competes for the same primer set in the conventional PCR assay, allowed for detection and quantification of R. secalis fungal biomass. In order to generate a standard calibration curve, DNA prepared from infected seeds with different levels of R. secalis infection was subjected to competitive PCR assay. The resulting PCR product ratio for each PCR reaction ( R. secalis -amplified DNA/internal control template-amplified DNA) increased proportionally with increasing levels of infected seed DNA in the reaction mixture. Naturally infected seed lots collected from 1995 to 1999 were used to demonstrate the potential of the competitive PCR assay as an alternative seed health testing method. The results from this competitive PCR assay were compared with those from conventional visual disease assessment and an agar plate assay. Although relatively good correlation between visual disease assessment and the competitive PCR was found in the case of artificially mixed seed samples, there was poor correlation in the experiments using naturally infected seed samples.     

Temporary partial breakdown of Mlo-resistance in spring barley by the sudden relief of soil water stress

Seedlings of spring barley cultivars expressing mlo genes for resistance to mildew (Erysiphe graminis f.sp. hordei) in different genetic backgrounds were subjected to degrees of water stress under different nutrient and soil conditions. The temporary breakdown in the host of the expression of Mlo-resistance, reported occasionally under field conditions, was successfully simulated when stress treatments were relieved. Differences in the degree of temporary breakdown were attributable to nutrient status, soil compaction and genetic background of the mlo gene.     

Leaf blotch severity on spring barley infected by isolates of Rhynchosporium secalis under different temperature and humidity regimes

The infection efficiency and severity of leaf blotch on spring barley inoculated with three pathotypes of Rhynchosporium secalis from central Norway were studied under different temperature and humidity regimes. Seedlings of the cultivar Arve were subjected to two constant temperatures, 13° or 18°C. Dry periods of 8 h or longer before or after a wet period of 4 h, carried out in the first 48 h postinoculation, reduced disease severity assessed 16 days after inoculation. The effect of dry periods of up to 24 h was nullified when plants were subjected to high humidity for 48 h after the dry treatment. The disease developed most rapidly when the wet period was 48 h and the temperature 18°C. At or near the optimum temperature for R. secalis (18°C), leaf wetness duration as short as 2 h resulted in considerable disease. Isolates reacted differently to temperature. The most aggressive isolate caused severe disease irrespective of temperature (56–70% of the leaf area infected); however, disease severity caused by the least aggressive isolate was significantly higher at the optimum temperature compared with a lower temperature (13°C). This information can facilitate evaluation of weather data in relation to predicting leaf blotch for advisory purposes.     

Sequence analysis and classification of apparent recombinant begomoviruses infecting tomato in the nile and mediterranean basins

ABSTRACT Numerous whitefly-transmitted viral diseases of tomato have emerged in countries around the Nile and Mediterranean Basins the last 20 years. These diseases are caused by monopartite geminiviruses (family Gemini viridae) belonging to the genus Begomovirus that probably resulted from numerous recombination events. The molecular biodiversity of these viruses was investigated to better appreciate the role and importance of recombination and to better clarify the phylogenetic relationships and classification of these viruses. The analysis partitioned the tomato-infecting begomoviruses from this region into two major clades, Tomato yellow leaf curl virus and Tomato yellow leaf curl Sardinia virus. Phylogenetic and pairwise analyses together with an evaluation for gene conversion were performed from which taxonomic classification and virus biodiversity conclusions were drawn. Six recombination hotspots and three homogeneous zones within the genome were identified among the tomatoinfecting isolates and species examined here, suggesting that the recombination events identified were not random occurrences.     

Existing infection with <Emphasis Type="Italic">Rhynchosporium secalis</Emphasis> compromises the ability of barley to express induced resistance

It has been suggested that if plants in the field are already induced, their ability to further enhance induced resistance might be compromised. This was examined in barley by inoculating the first two leaves with Rhynchosporium secalis prior to treatment of leaves three and four with an elicitor combination, followed by inoculation with R. secalis. The elicitor combination used consisted of acibenzolar-S-methyl, β-aminobutyric acid, and cis-jasmone, which was shown previously to provide higher levels of disease control in barley than any of the components used individually. The elicitor combination reduced infection by R. secalis, and led to an up-regulation of PR1-b, a marker gene for systemic acquired resistance, and increased activities of the defence-related enzymes cinnamyl alcohol dehydrogenase (CAD), peroxidase (POX), and β-1,3-glucanase. It also led to down-regulation of LOX2, a gene involved in biosynthesis of jasmonic acid. In plants where the first two leaves were inoculated with R. secalis prior to treatment of leaves three and four with elicitor, these increased defence responses did not occur, and control of R. secalis infection on leaves three and four was also reduced. These results suggest that, at least in young barley plants, prior infection with R. secalis compromises their ability to respond effectively to elicitors. The results might help to explain the relatively poor performance of induced resistance in the field, particularly in cereals, compared to plants grown under controlled conditions.