Cellular characteristics of temporary partial breakdown of mlo- resistance in barley to powdery mildew

When water-stress is relieved, powdery mildew (Erysiphe graminis f.sp. hordei) infection increases on both Mlo -susceptible and mlo -resistant spring barley cultivars. The breakdown is temporary and is determined by the genetic background rather than the specific resistance allele. The relief of water-stress time-point for maximum mildew infection frequency on mlo -resistant barley is approximately 7 h post-inoculation. The degree of breakdown varies with epidermal cell type; increased infection frequency is greatest on short followed by long and adjacent epidermal cell types, rather than on the stomatal subsidiary cells with which occasional colonies are typically associated. Infection frequency on the short cells of mlo -resistant cv. Atem increases from less than 1% under a non-stressed watering regime to more than 10% after a relief of water-stress at 7 h post-inoculation. Following haustorium formation, colonies develop and reach sporulation within 7 days post-inoculation. In host epidermal cells, the extent of cross-linked protein at interaction sites is reduced under conditions of water-stress or stress-relief. Cross-linked protein is reduced in terms of the frequency of occurrence at primary germ tube interaction sites 7 h post-inoculation and the deposition size at appressorial germ tube interaction sites 24 h post-inoculation. This indicates a delayed or reduced defence response during the recovery period. These data demonstrate a differential cellular response to powdery mildew in barley genotypes prone to resistance expression breakdown following relief of water-stress.     

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.     

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.     

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.     

Immigration of the barley mildew pathogen into field plots of barley

Immigration of the barley powdery mildew pathogen ( Erysiphe graminis f.sp. hordei ) into field plots of the spring barley variety Tyra (carrying the resistance allele Mla1 ) was investigated. Spores were trapped from the top of the plot canopies, as well as from control plots of wheat with no barley nearby. Comparison of the frequencies of virulent and avirulent single-colony isolates showed that the amount of immigration, relative to the amount of inoculum being produced within the plot, reduced very rapidly, until it could not be detected in the middle of the growing season (mid-June).     

Antioxidant,ethylene and membrane leakage responses to powdery mildew infection of near-isogenic barley lines with various types of resistance

Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related near-isogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5–7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3–5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H₂O₂ were observed in the inoculated resistant plants. In contrast, H₂O₂ content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.     

大麦白粉菌种群毒性监测及抗性材料鉴定

2005和2006年从我国冬大麦区采集和分离大麦白粉菌单胞菌株729个,利用Pallas近等基因系进行致病型鉴定和群体毒性频率分析.同时,利用分离得到的不同致病型菌株,通过抗谱分析的方法鉴定了328份大麦品种(系)的白粉病抗性和抗病基因.结果显示:大麦白粉菌群体对抗病基因Mlal Mla(A12)、Mla3、Mla6 Mla14、Mla7 Mla(No3)、Mla7 Ml(Lg2)、Mla9 Mlk、Mla9、Mlal3 MlaRu3、Mlpl、Mlg(Cp)和mlo5的毒性频率为0;对Mla12 MlaEm2、Mla7 Mlk、Mlat Mla8、Mla10MlaDu2和Mlk1的毒性频率很低,分别为0.1%、0.4%、0.9%、2.8%和4.2%.两年共鉴定出不同的致病型21个,致病型000、001和003在两个年度皆为优势致病型.所鉴定的328份材料绝大多数感病,仅37份抗病材料,能明确推导出抗白粉病基因的品种(品系)很少,这些品种(品系)含有的抗白粉病基因为Mr(Bw)Mla8、Mlg、Mira Mla8、Mla9 Mla1、Mla Mla(A12)和mlo5.     

Effects of impacting rain drops on the growth and development of powdery mildew fungi

The effects of rain on different stages of powdery mildew development were investigated quantitatively for Erysiphe graminis on barley, Erysiphe pisi on pea and Sphaerotheca pannosa on rose. Water drops 4 mm in diameter released from a height of 2·5 m onto target leaves reduced the numbers of conidia on the impacted surfaces and inhibited subsequent mildew development, the effects declining as the interval between inoculation and impaction increased. Simulated rainfall applied after inoculation reduced the numbers of conidia on leaves and inhibited mildew development, the effect declining as the interval between inoculation and treatment lengthened. Simulated rainfall applied to infected plants reduced their capacity to inoculate other plants, the effect declining as the interval between treatment and use as inoculum lengthened. Controlled exposure of infected peas and roses to natural rain reduced the numbers of conidia on leaves and inhibited subsequent mildew development, especially on upper leaf surfaces.     

Virulence patterns in populations of Erysiphe graminis f.sp. hordei in Europe in 1986

A total of 3166 single colony progeny of Erysiphe graminis f.sp. hordei , constituting random samples of the aerial inoculum from 68 regions of Western Europe, were analysed in the laboratory for virulence to barley resistance genes Mlra, Mlg, Mla6+Mlg. MILa, Mla7+Mlk, Mlall, Mla9, MlaI3 + MlRu3. Mlal, Mla3 and mlo. No isolate virulent to mlo was detected. Distinct regional patterns were observed for the frequencies of most of the other virulence genes. They stressed the role of selection exerted by the cultivars grown, as well as the effect of wind dissemination of mildew populations. These two phenomena are particularly well exemplified by the data on early stages in the evolution of the newly emerging Val3 virulence in Denmark. In contrast with the preceding evolution, there was a considerable decrease of the frequency of a broadly distributed virulence ( Va6); the most probable reasons for this are outlined.     

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.     

Effects of barley genotype on induced resistance to powdery mildew

Infection of barley by an avirulent isolate of Erysiphe graminis f.sp. hordei reduces the development of powdery mildew disease by a virulent isolate, inoculated subsequently. This phenomenon, known as induced resistance, was studied in three sets of near-isogenic lines of barley, each of which included varieties with four different race-specific resistance genes. The extent of induced resistance, expressed as a reduction in the number of colonies formed, differed in lines carrying different race-specific resistance genes, being most effective in lines with Mla7 , followed by Mth , then Mla6 and Mla13. These differences may be related to the extent of the hypersensitive response to infection by avirulent spores. A further effect of induced resistance in reducing sporulation, taking the number of colonies into account, was least effective in Mth lines, with the other lines ranking as they did when induced resistance was expressed as colony numbers. By contrast, the genetic backgrounds of the three near-isogenic sets had similar effects on the fractional reduction of colony number by induced resistance. The effect of induced resistance on sporulation was strongest in the most resistant background, and weakest in the most susceptible.     

Variation in adult plant resistance to powdery mildew in spring oats under field and laboratory conditions

Ten oat genotypes were assessed for adult plant resistance (APR) to powdery mildew ( Erysiphe graminis f.sp. avenae ) in small field plots and on detached leaf segments in the laboratory using glasshouse-grown plants. Significant genotype differences were observed in the field and on leaf segments for latent period, infection frequency and spore production, components used to assess APR. The genotype OM1711, which was selected because it expressed a higher level of mildew resistance than its most resistant parent, cv. Maldwyn, showed this transgressive resistance in both field and laboratory tests. By contrast, the genotype OM1387, also selected for high mildew resistance, only showed this enhancement in the field. It is concluded that quantitative assessment of mildew in detached leaves is not sufficiently precise to be used alone in selecting segregants for high levels of APR, and that field data are also needed.     

Comparison of resistance-inducing abilities of virulent and avirulent races of Erysiphe graminis f.sp. hordei and a race of Erysiphe graminis f.sp. tritici in barley

Resistance to powdery mildew was induced in barley by preinoculation with virulent and avirulent races of barley powdery mildew ( Erysiphe graminis f.sp. hordei ), and with a race of wheat powdery mildew ( E. graminis f.sp. tritici ). Four inducer densities were tested in 13 different induction periods between 1 and 24 h. Generally, the resistance induced by barley powdery mildew increased up to 10-12 h of induction and was maintained in longer induction periods. The inducing abilities of virulent and avirulent races could not be distinguished up to 10-12 h of induction, after which the inducing ability of avirulent races increased significantly in relation to virulent races. Wheat powdery mildew was able to induce more resistance than barley powdery mildew in induction periods up to 8 h. In a single inoculation procedure the number of haustoria developing from virulent barley powdery mildew decreased as inoculum density increased. The effect was ascribed to induction of resistance. This reduction of infection efficiency in the compatible interaction was compared to induced resistance. However, the inoculum density needed for 50% resistance induction in the double inoculation procedure was approximately 40 times higher than the inoculum density needed for 50% reduction in infection efficiency in the single inoculation procedure.     

Breakdown of Pl2, a major gene of resistance to apple powdery mildew, in a French experimental orchard

Pl2 , a major resistance gene to powdery mildew originating from Malus zumi , is mainly used in apple-breeding programmes. In an experimental orchard in France, an increase of susceptibility to powdery mildew was observed on apple genotypes carrying Pl2 . This increase of susceptibility could not be explained by an effect of the age of the trees, or by an effect of the amount of inoculum on expression of the resistance Pl2 . It was demonstrated by tests of pathogenicity in controlled conditions that isolates of Podosphaera leucotricha sampled in this orchard were virulent to Pl2 . The appearance of virulent isolates within 6 years after planting apple genotypes selected for the presence of Pl2 resulted in a breakdown of the resistance. Ten years after planting, the percentage of genotypes that were still resistant to powdery mildew varied between 2 and 56%, depending on the progenies. This suggested that additional quantitative resistance genes may influence the durability of the resistance Pl2 .     

Genetic control of the resistance of Erysiphe graminis f.sp. hordei to five triazole fungicides

The genetics of resistance to ergosterol demethylation inhibitor (DMI) fungicides of the triazole (conazole) group was examined in a cross between two isolates of the barley powdery mildew fungus, Erysiphe graminis (=  Blumeria graminis ) f.sp. hordei . One isolate, E1, was previously identified as being resistant to the triazole fungicide triadimenol, while the other, HL3/5, was sensitive. The 56 progeny tested were classified into two distinct groups, either being resistant to triadimenol, like E1, or sensitive, like HL3/5. The segregation ratio was not significantly different from 1:1, consistent with responses to triadimenol being controlled by a single gene. In further tests with cyproconazole, epoxiconazole, propiconazole and tebuconazole, all the progeny classified as resistant to triadimenol were also more resistant to each of these other triazole fungicides than were any of the triadimenol-sensitive progeny. This is consistent with the triadimenol resistance allele also conferring cross-resistance to the other triazoles. The ratio between the responses of the resistant and sensitive progeny (the resistance factor, RF) was greatest for triadimenol, followed by tebuconazole, propiconazole, epoxiconazole and cyproconazole, in that order. The RF for triadimenol was much greater when the fungicide was applied as a seed treatment than when it was sprayed. Five isolates, covering the five levels of responses to triadimenol identified previously in the UK population of E. graminis f.sp. hordei , were used as standards; a triadimenol-sensitive isolate and one with the lowest level of resistance were sensitive to all four of the other fungicides, while three isolates with higher levels of triadimenol-resistance were also more resistant to the other chemicals.     

Spectral signatures in the UV range can be combined with secondary plant metabolites by deep learning to characterize barley–powdery mildew interaction

In recent studies, the potential of hyperspectral sensors for the analysis of plant–pathogen interactions was expanded to the ultraviolet range (UV; 200–380 nm) to monitor stress processes in plants. A hyperspectral imaging set-up was established to highlight the influence of early plant–pathogen interactions on secondary plant metabolites. In this study, the plant–pathogen interactions of three different barley lines inoculated with Blumeria graminis f. sp. hordei (Bgh, powdery mildew) were investigated. One susceptible genotype (cv. Ingrid, wild type) and two resistant genotypes (Pallas 01, Mla1- and Mla12-based resistance and Pallas 22, mlo5-based resistance) were used. During the first 5 days after inoculation (dai) the plant reflectance patterns were recorded and plant metabolites relevant in host–pathogen interactions were studied in parallel. Hyperspectral measurements in the UV range revealed that a differentiation between barley genotypes inoculated with Bgh is possible, and distinct reflectance patterns were recorded for each genotype. The extracted and analysed pigments and flavonoids correlated with the spectral data recorded. A classification of noninoculated and inoculated samples with deep learning revealed that a high performance can be achieved with self-attention networks. The subsequent feature importance identified wavelengths as the most important for the classification, and these were linked to pigments and flavonoids. Hyperspectral imaging in the UV range allows the characterization of different resistance reactions, can be linked to changes in secondary plant metabolites, and has the advantage of being a non-invasive method. It therefore enables a greater understanding of plant reactions to biotic stress, as well as resistance reactions.     

Differential effects of phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, and energetic metabolism inhibition on resistance of appropriate host and nonhost cereal-rust interactions

ABSTRACT Effects of phenylpropanoid and energetic metabolism inhibition on resistance were studied during appropriate host and nonhost cereal-rust interactions. In the appropriate barley-Puccinia hordei interaction, phenylalanine ammonia lyase (PAL) and cinnamyl alcohol dehydrogenase (CAD) inhibition reduced penetration resistance in two genotypes, suggesting a role for phenolics and lignins in resistance. Interestingly, penetration resistance of the barley genotype 17.5.16 was not affected by phenylpropanoid biosynthesis but penetration resistance was almost completely inhibited by D-mannose, which reduces the energy available in plant host cells. This suggests a parallel in the cellular basis of penetration resistance between 17.5.16 rust and mlo barleys powdery mildew interaction. Results revealed differing patterns of programmed cell death (PCD) in appropriate versus nonhost rust interactions. PAL and CAD inhibitors reduced PCD (hypersensitivity) in appropriate interactions. Conversely, they had no effect in PCD of wheat to P. hordei; whereas D-mannose dramatically reduced nonhost resistance and allowed colony establishment. The differential effects of inhibitors in the expression of the different resistances and the commonalities with the cereal-powdery mildew interaction is analyzed and discussed.     

Comparison of mobile and stationary spore-sampling techniques for estimating virulence frequencies in aerial barley powdery mildew populations

Gene frequencies in samples of aerial populations of barley powdery mildew ( Erysiphe graminis f.sp. hordei ), which were collected in adjacent barley areas and in successive periods of time, were compared using mobile and stationary sampling techniques. Stationary samples were collected from trap plants in three periods within 1 week at a distance of more than 1000 m from the nearest barley field. At four dates within the same 8-day period, other samples were collected by a mobile spore trap along four sampling routes of a total distance of 130 km around the stationary stand of exposure. The samples were characterized by virulence genotypes defined according to infection types on 12 near-isogenic barley lines, and frequencies of single virulence genes were subsequently calculated. The three samples collected at the stationary site differed significantly with respect to allele frequencies at three loci. The main wind direction was different in the three sampling periods, implying different powdery mildew sources. For the mobile exposure, the differences between routes were not significant for any locus. However, the routes differed most for the loci under direct selection by host resistances genes, indicating a different distribution of source varieties along routes. There was no difference between allele frequencies at different dates, indicating that the proportions of spores from different source varieties were similar at these dates. In conclusion, samples collected by the stationary technique will mainly reflect the source varieties present in the local area, whereas samples collected by the mobile spore trap will mainly reflect sources close to the sampling route. Therefore, sampling sites as well as sampling routes should be defined such that source varieties are representative for the overall varietal distribution in the survey region considered.     

Differential expression of resistance to powdery mildew incited by race 1 or 2 ofsphaerotheca fuliginea inCucumis melo genotypes at various stages of plant development

Thirty-nine genotypes ofCucumis melo (plant introduction entries, open-pollinated cultivars and F₁ hybrids) were evaluated for resistance to powdery mildew under either natural field conditions or artificial inoculation in growth chambers at the cotyledonary stage and the 2-true-leaf stage. Results confirmed that susceptibility in cotyledons was not necessarily associated with susceptibility in either true leaves in growth chambers or adult plants in the field. However, resistance at the 2-true-leaf stage in growth chambers was highly correlated with resistance of field-grown plants. Results also showed that 20 muskmelon genotypes resistant to race 1 at the cotyledonary stage were also resistant at the 2-leaf-stage and as adult plants in the field. The same was true for ten genotypes with race 2 inoculations. Because muskmelon genotypes expressing resistance in cotyledons were also resistant in true leaves in growth chambers or the field, the use of plants at the cotyledonary stage is recommended for screening for powdery mildew resistance caused by race 1 or race 2 ofS. fuliginea. When cotyledons are susceptible, screening should be done at the 2-true-leaf stage.     

Quantitative trait Loci mapping for adult-plant resistance to powdery mildew in bread wheat

ABSTRACT Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a major disease to wheat (Triticum aestivum) worldwide. Use of adult-plant resistance (APR) is an effective method to develop wheat cultivars with durable resistance to powdery mildew. In the present study, 432 molecular markers were used to map quantitative trait loci (QTL) for APR to powdery mildew in a doubled haploid (DH) population with 107 lines derived from the cross Fukuho-komugi x Oligoculm. Field trials were conducted in Beijing and Anyang, China during 2003-2004 and 2004-2005 cropping seasons, respectively. The DH lines were planted in a randomized complete block design with three replicates. Artificial inoculation was carried out in Beijing with highly virulent isolate E20 of B. graminis f. sp. tritici and the powdery mildew severity on penultimate leaf was evaluated four times, and the maximum disease severity (MDS) on penultimate leaf was investigated in Anyang under natural inoculation in May 2004 and 2005. The heritability of resistance to powdery mildew for MDS in 2 years and two locations ranged from 0.82 to 0.93, while the heritability for area under the disease progress curve was between 0.84 and 0.91. With the method of composite interval mapping, four QTL for APR to powdery mildew were detected on chromosomes 1AS, 2BL, 4BL, and 7DS, explaining 5.7 to 26.6% of the phenotypic variance. Three QTL on chromosomes 1AS, 2BL, and 7DS were derived from the female, Fukuho-komugi, while the one on chromosome 4BL was from the male, Oligoculm. The QTL on chromosome 1AS showed high genetic effect on powdery mildew resistance, accounting for 19.5 to 26.6% of phenotypic variance across two environments. The QTL on 7DS associated with the locus Lr34/Yr18, flanked by microsatellite Xgwm295.1 and Ltn (leaf tip necrosis). These results will benefit for improving powdery mildew resistance in wheat breeding programs.