Evidence for Resistance-inducing Compounds in Conidia of Erysiphe graminis f.sp. hordei

Preparations of Erysiphe graminis f.sp. hordei conidia were spray-applied to the first leaf of barley plants which were subsequently challenge inoculated with virulent powdery mildew. The powdery mildew reducing effect of the preparations was assessed by scoring the outcome of the challenge inoculation. Homogenates of ungerminated conidia, germinated conidia, and methanol-water extracts of germinated conidia reduced the number of powdery mildew colonies. Cell wall fragments from ungerminated conidia, germinated conidia, and conidial germination fluid obtained from conidia germinated in aqueous suspension did not reduce the number of powdery mildew colonies. Microsconical analysis of the infection course following challenge inoculation indicated that the powdery mildew reducing effect is due partly to induced resistance.     

The Effects of Brown Rust and Phosphate on Cold Acclimation in Winter Barley

The green leaf area of winter barley, cv. Sonja, sampled fromthe field at different times during winter was always greatestin plants grown at high soil phosphate and smallest in plantsgrown at low soil phosphate, and at each fertilizer level wasgreater in healthy plants than in plants infected by rust (Pucciniahordei). In leaves that survived the coldest period of winter,the percentage area that was damaged was increased by rust infectionwhich prevented the ameliorating effects of high soil P. Rustand low P interacted to reduce the increases in leaf area andshoot d. wt that occurred when higher temperatures prevailedin spring. Under controlled conditions in the laboratory, phosphate reducedthe injury suffered when plants not acclimated to low temperatureswere exposed to freezing conditions, but this effect was removedby rust infection. After rust infection, freezing temperatureswere damaging even to acclimated plants, particularly if grownwith low soil P. Evidence of visible symptoms, and quantitativemeasurements of electrolyte efflux from intact leaves, chlorophyllfluorescence in vivo, and ethane and ethylene evolution fromcold-acclimated plants, showed that infection raised the minimumtemperature at which tissues could survive without injury. Infectedleaves were more sensitive to low temperature post-sporulationthan presporulation. Measurements of electrolyte efflux andchlorophyll fluorescence on plants growing under cold conditionsshowed that infection inhibited the processes of acclimationto low temperatures. Winter barley, Puccinia hordei, injury, low temperature, acclimation     

Germination of Erysiphe gratninis f.sp. hordei conidia on barley leaves

Germination of Erysiphe graminis f.sp. hordei conidia on leaves of several barley cultivars was studied in the laboratory. On both detached leaves and intact plants, within 48 h of inoculation a higher proportion of conidia had germinated on the basal and middle portions of the adaxial leaf surface than on the corresponding portions of the abaxial surface. Such differences between surfaces were not observed near the leaf tip. Similar results were obtained with all the cultivars and growth stages tested, and with five isolates of E. graminis, and are consistent with the observation that there is usually less powdery mildew on the abaxial than the adaxial surface of barley leaves. With most of the barley genotype/mildew isolate combinations tested, within 48 h of inoculation higher proportions of conidia germinated on seedlings and juvenile plants than on older plants. Inherited characteristics which affect spore germination on the leaf surface may be important factors in the development of adult-plant resistance of barley to powdery mildew, particularly in certain genotypes.     

Synthesis and antifungal activity of six benzylic diamines

Six benzylic diamines were synthesised and examined for antifungal activity. Four of the compounds, KB 2, KB 4, KB 5 and KB 6, reduced radial growth of the oat leaf stripe pathogen Pyrenophora avenae, the largest reduction obtained with 25 μM KB 4, which reduced radial growth by 47%. Surprisingly, these four amines had no effect against infection of barley seedlings with the powdery mildew fungus Erysiphe graminis f.sp. hordei. Instead, two different amines, KB 1 and KB 3, reduced powdery mildew infection on barley. The greatest reduction was obtained with 25 μM KB 3, which reduced mildew infection by 69%. All of the amines which exhibited antifungal or fungicidal properties perturbed polyamine formation as measured by the incorporation of labelled ornithine into polyamines.     

Translocation of carbon in powdery mildewed barley

This paper compares translocation in healthy and powdery mildew (Erysiphe graminis f. sp. hordei, race CR3) infected barley (Hordeum vulgare, variety Manchuria). The sink-like properties of the powdery mildew infection were used to determine what effect imposing a sink in the midst of normal source tissue (mature primary leaf) had on the translocation process. The pattern of translocation was determined by monitoring the movement of ¹⁴C which was photosynthetically incorporated from ¹⁴C either by the primary or second leaf. In the healthy primary leaf of barley, ¹⁴C fixed in the tip section of the blade was preferentially translocated to the root, whereas ¹⁴C fixed in the basal section was primarily translocated to the shoot. When a sporulating powdery mildew infection was present in the mid-section of the primary leaf, ¹⁴C fixed in that section or in the acropetal healthy tip section readily accumulated in the infection area. Labeled carbon fixed in the healthy basal section was translocated into the other parts of the plant with only a small fraction moving acropetally into the infected mid-section. The ¹⁴C fixed by the second leaf was translocated to the root and younger shoot with very little entering the primary leaf. The presence of the mildew infection did not alter this pattern.     

Assimilatverteilung in Sommergerstensorten mit unterschiedlicher Resistenz gegenüber dem Echten Mehltau (Erysiphe graminis f. sp. hordei)

Distribution of assimilates in cultivars of spring barley with different resistance against powdery mildew (Erysiphe graminis f. sp. hordei) Transport and distribution of radioactive labelled assimilates in spring barley cultivars with different degrees of resistance to powdery mildew were studied after ¹⁴CO₂-treatment of single leaves. Plants of the cultivars ‘Amsel’ (susceptible), ‘Asse’ (adult plant resistant), and ‘Rupee’ (resistant) were analyzed at the vegetative growth stage (5. leaf unfolded) and the generative growth stage (anthesis). At the vegetative growth stage the assimilate export from the mildew inoculated 5. leaf of ‘Amsel’ and ‘Rupee’ is decreased; in ‘Asse’, there is no considerable change of assimilate distribution due to infection. At the generative growth stage the assimilate export from the infected flag leaf of ‘Amsel’ is reduced when the fungus, is sporulating. In the cultivar ‘Asse’ the assimilates are bound at the infection site until the seventh day after inoculation, then the transport of assimilates to the ear is increased. In ‘Rupee’ mildew inoculation causes an enhanced assimilate transport to the ear. The changes in assimilate distribution due to mildew inoculation are discussed with respect to the different types of host-parasite-interactions and the source-sink-activities in the different cultivars.     

Effects of soil water level on the development of adult plant resistance to powdery mildew in barley

Barley grown in dry soil developed greater adult plant resistance (APR) to powdery mildew (Erysiphe graminis DC. f. sp. hordei Mérat) than barley grown in wet soil. Conidial germination and appressorium formation were less, and fungal development between formation of appressoria and elongating secondary hyphae on upper leaves was inhibited, when adult plants were grown in dry soil. Mildew colonies expanded more slowly on leaves of adult plants than on leaves of seedlings, especially if adult plants had grown in dry soil. APR was reduced if plants, previously grown in dry soil, were well watered more than 32 h before inoculation. Conidia originating from plants grown in dry soil had a lower solute potential and greater ability to infect plants grown in dry but not wet soil than conidia originating from plants grown in wet soil. APR could not be attributed simply to increased cell wall or cuticle thickness, nor to lowered leaf solute potentials, as has sometimes been suggested for powdery mildew diseases. Increasing plant age and water stress induced increases in cell wall and cuticle thickness, but these changes did not always coincide with changes in disease resistance. Increasing plant age and water stress also lowered leaf solute potentials but fungal solute potentials were lower than leaf solute potentials and, more importantly, were lower than leaf water potentials. Thus, fungal growth was not limited by the availability of water from the host during penetration and hyphal establishment. It is suggested that resistance levels may be determined not by the thickness of epidermal structures, nor by lowering of solute potential per se, but by specific substances harmful to the fungus which accumulate in either cell wall, cuticle or sap, and whose concentration is dependent on the age and water stress of leaves.     

Controlled environment experiments on epidemics of barley mildew in different density host stands

Summary Controlled environment experiments on small epidemics of powdery mildew of barley, an air-borne disease caused by Erysiphe graminis f.sp. hordei, indicated that there was a direct linear relationship between host density and the rate of increase of disease within populations. Under the particular experimental conditions used, the overall infection rate was almost doubled (from 0.39 to 0.75 per unit per day) following an increase in density from 31 to 115 host units per m². In separate experiments these overall epidemic rates were partitioned into two separate components related to inoculum transmission between plants and inoculum transmission within plants.     

Ribulose Bisphosphate Carboxylase Protein and Enzymes of CO2 Assimilation in Barley Infected by Powdery Mildew (Erysiphe graminis hordei)

Effects were examined of barley powdery mildew (Erysiphe graminis f. sp. hordei) on the major leaf protein, ribulose-l,5-bisphosphate carboxylase (RuBPCase), and other enzymes of CO₂ assimilation, phosphoenolpyruvate carboxylase (PEPcase) and malic enzymes, and enzymes associated with RuBPCase in the reductive pentose phosphate pathway. Activity of RuBPCase per unit fresh weight of leaf was inhibited by infection from the first sample, 3 days after inoculation, to the last sample, 24 days after inoculation, when healthy control leaves were visibly senescing. The inhibition occurred because the amount of RuBPCase protein (measured specifically by an immunological technique) was reduced from 6 days after inoculation and because activity per unit protein declined from 3 days until 21 days after inoculation, at which time there was little protein remaining. Activity of PEPcase per unit fresh weight of leaf was initially stimulated by infection but, thereafter, it was inhibited. Inhibition, like that also affecting malic enzymes (NAD) and (NADP), 3-phosphoglycerate kinase, and glyceraldehyde-3-phosphate dehydrogenases (NAD) and (NADP), was associated with a decline in amounts of total soluble minus RuBP Case protein per unit fresh weight of leaf. Reduced amounts of leaf protein may be associated with reduced nitrate uptake by roots and fungal demand for nitrogen in mildewed plants.     

Induction of Systemic Resistance to Tobacco Powdery Mildew by Tobacco Mosaic Virus, Tobacco Necrosis Virus or Ethephon

Local infections of either TMV or TNV in tobacco plants cv. Havana 425 (hypersensitive to TMV) proved effective in inducing systemic resistance to subsequent inoculation with the powdery mildew fungus Erysiphe cichoracearum DC. The proportion of leaf surface invaded by this pathogen and the amount of conidia it produced were both significantly lower in virus inoculated plants than in non-inoculated controls. However, the decrease in sporulation rate was less regularly observed than the reduction in leaf area infected. TMV was more effective than TNV in protecting tobacco plants from powdery mildew. E. cichoracearum is thus added to the list of challenge pathogens to which TMV or TNV are known to induce resistance in the host plants. Necrotic lesions caused to the leaves by local treatment with Ethephon (an ethylene-releasing compound) also conferred to tobacco some degree of systemic resistance to the same fungal pathogen, more frequently visible as a reduction of leaf area invaded. The protection due to the Ethephon lesions was in present experiments less marked than that of TMV. No effects against subsequent powdery mildew infection were obtained when point freeze necrotic lesions were provoked on the plants.     

Postulation of resistance genes to barley diseases in heterogeneous varieties

Resistance to causal agents of diseases is an important varietal characteristic that influences the management practice of crop plants and thus production costs of commodities. At present, almost all European barley varieties possess at least one major gene for resistance to powdery mildew. After hybridizing selected parental varieties, resistance genes often segregate in subsequent generations and, therefore, some varieties comprise lines that differ in the number or combinations of resistance genes. The objective of this research was to describe the various methods available for postulating resistance genes to pathogens in heterogeneous varieties using resistance to powdery mildew of barley as an example. Four spring barleys (‘Orbit’, ‘Malva’, ‘Tocada’ and CLE 233), and a six-row variety of winter barley, F 12872, were screened. For postulating resistance genes, several testing procedures and many Blumeria graminis f.sp. hordei isolates were used. Minimum amounts of seed were determined and different methods of obtaining homogeneous seed samples from heterogeneous varieties were compared. It was found that ‘Orbit’ and ‘Malva’ are composed of three and ‘Tocada’, CLE 233 and F 12872 of two lines with different resistances to powdery mildew. Problems of postulating resistance genes in heterogeneous varieties and the advantages of testing leaf segments instead of young plants are discussed.     

Analysis of the Effects of Erysiphe graminis DC. on the Growth of Barley

Three barley varieties, Plumage Archer, Proctor, and Haisa II,inoculated soon after emergence with Erysiphe graminis, thecause of powdery mildew, grew and yielded less than barley repeatedlysprayed with lime sulphur. Size of roots was affected more than tops, so the dry-weightratio of root/total plant was decreased. The smaller tops reflectedfewer shoots which, later, were also smaller than on sprayedplants. Decreases in leaf area paralleled decreases in dry weight oftops. Powdery mildew affected the balance between the absorbingsystem and the assimilating and transpiring system by decreasingthe average dry weight of roots per unit leaf area from 0.92to 0.63 mg./cm². Disease also decreased the efficiency of unitleaf area; from 12 to 68 days after inoculation, the mean netassimilation rate was 226.6 in sprayed controls and 166.0?8.6m.g./dm.²/week in the inoculated series. Powdery mildew retarded stem elongation and possibly causedstunting. Fewer and smaller ears were produced by inoculatedthan by sprayed plants.     

Development of powdery mildew on leaves of several barley varieties at different growth stages

On detached leaves and intact plants of several barley varieties at different growth stages, lower percentages of germinated conidia of Erysiphe graminis f.sp. hordei penetrated the host and initiated infection on the abaxial than adaxial surface. More and larger E. graminis colonies developed on the adaxial surface and these comprised more densely packed hyphae and produced more conidiophores than did colonies on the abaxial surface. These results are consistent with the observation that there is usually more powdery mildew on the adaxial than abaxial surface of barley leaves in the field. Smaller proportions of germinated E. graminis conidia penetrated and infected the host on leaves of adult or near-adult plants than on those of seedlings or juvenile plants. Older plants also supported fewer, smaller and less dense colonies with less sporulation than young plants. The effects of growth stage of the host plant on development of powdery mildew were much greater in some barley varieties, and with some E. graminis isolates, than others.     

The effects of host carbon dioxide, nitrogen and water supply on the infection of wheat by powdery mildew and aphids

In two experiments, winter wheat (Triticum aestivum cv. Cerco) was grown in 350 (ambient) and 700 μmol mol^-1 (elevated) atmospheric CO₂ concentrations. In the first experiment, plants were grown at five levels of nitrogen fertilization, and in the second experiment, plants were grown at three levels of water supply. All plants were infected with powdery mildew, caused by the fungus Erysiphe graminis. Plants grown in elevated atmospheric CO₂ concentrations had significantly reduced % shoot nitrogen contents and significantly increased % shoot water contents. At elevated atmospheric CO₂ concentrations, where plant nitrogen content was significantly reduced, the severity of mildew infection was significantly reduced, and where host water content was significantly increased, the severity of mildew infection was significantly increased. In a moderate water supply treatment, the plants grown in elevated atmospheric CO₂ concentrations had significantly reduced nitrogen contents (9·9%) and significantly increased water content (4%), the amount of mildew infection was unchanged. The severity of mildew infection appeared to be more sensitive to host water content than to host nitrogen content.     

Effects of Mildew and Nitrogen on Grain Yield of Barley Artifically Infected in the Field

Powdery mildew was artificially introduced into field plotsof barley given two amounts of nitrogen fertilizer. In Tyraand Triumph, grain dry matter and nitrogen yield of healthyplots increased in response to nitrogen fertilizer. In mildewedplots increases in response to nitrogen did not occur in Triumph,despite low levels of mildew infection. Mildew reduced grainyield and nitrogen content at both levels of nitrogen in bothvarieties. Hordewm vulgare L., barley, Erysiphe graminis, powdery mildew, nitrogen fertilizer, grain yield     

Synthesis and antifungal activity of two novel spermidine analogues

Two spermidine analogues were synthesised and examined for antifungal activity. Both compounds used as 1 mM post-inoculation sprays reduced infection of barley seedlings by the powdery mildew fungus, Erysiphe graminis f.sp. hordei, infection of broad bean seedlings by the rust fungus, Uromyces viciae-fabae, and infection of apple seedlings by the powdery mildew fungus, Podosphaera leucotricha. Since these fungal pathogens cannot be cultured axenically, the effects of the two spermidine analogues on mycelial growth in vitro, as well as preliminary investigations on polyamine biosynthesis, were undertaken using the oat stripe pathogen, Pyrenophora avenae. Although neither compound affected radial growth of the fungus on plates, both analogues reduced fungal biomass in liquid culture substantially. The two spermidine analogues, used at a concentration of 1 mM, had no significant effect on the conversion of labelled ornithine into polyamines in P. avenae.     

Effects of fungicides and insecticides applied to spring barley sown on different dates in 1976-79

Factorial experiments in 1976–1979 investigated the effects of sowing date, fungicides (ethirimol seed treatments and tridemorph sprays) and insecticides (phorate applied to the soil, and menazon or dimethoate sprays) on powdery mildew, aphids, barley yellow dwarf virus (BYDV) and grain yield of spring barley (cv. Julia in 1976 and 1977; cv. Wing in 1978 and 1979). Late sowing usually increased the severity of powdery mildew, numbers of aphids and incidence of BYDV and generally decreased yield. Responses to pesticides were commonly greater on the late-sown than on the early-sown barley. Response to fungicides are principally attributed to the control of powdery mildew (Erysiphe graminis f. sp. hordei; the target species) but responses to insecticides cannot be attributed to virus control and seem unlikely to be due solely to control of aphids, whose numbers were relatively small. There were some effects of fungicides on aphids and insecticides on mildew, but they were inconsistent and too small to affect crop protection strategies.     

Field resistance of spring barley cultivars to powdery mildew in Lithuania

During vegetative period 2004–2005 powdery mildew (Erysiphe graminis DC. f. sp. hordei Em. Marchal) field resistance of spring barley cultivars was investigated at the Lithuanian Institute of Agriculture. The spring barley genotypes tested were Lithuania-registered cultivars, cultivars from genetic resources collection, and the new cultivars used for initial breeding. In total, 23 resistance genes were present in the 84 cultivars studied. Among mono-genes only mlo and 1-B-53 showed very high resistance. Slight powdery mildew necroses (up to 3 scores) formed on cultivars possessing these genes. The maximal powdery mildew (PM) severity reached a score of 8.5 and the area under disease progress curve (AUDPC) a value of 1216.8. The cultivars ‘Primus’, ‘Astoria’, ‘Power’, ‘Harrington’ and ‘Scarlett’ were the most resistant among the non mlo cultivars. Severity of PM on ‘Primus’ reached a score of 3.5 (3.0 of PM necrosis) in average, the other cultivars were diseased from 4.5 (3.0) to 5.0 (2.0). The AUDPC values for these cultivars except ‘Scarlett’ were the lowest (85.0–145.3) among the other cultivars. The highest contrast in development of the other leaf diseases was between highly resistant and susceptible to PM cultivar groups. The fast development of PM depressed development of the other diseases 4.7 times.     

Arbuscular mycorrhiza increased the activity of a biotrophic leaf pathogen – is a compensation possible?

Arbuscular mycorrhizal barley-plants were more susceptible to the obligate biotrophic shoot pathogen Erysiphe graminis f. sp. hordei. In experiments under greenhouse and open-air conditions on leaves of mycorrhizal plants, the sporulation rate of the mildew fungus was more than twice that on control plants. However, mycorrhizal plants suffered less than non-mycorrhizal plants in terms of grain number, ear yield and thousand-grain weight. Disease-yield-relationship analysis showed that the symbiosis neutralised the positive correlation between disease severity and yield loss (up to 25% infected leaf area tested). After mildew infection, nitrogen in ears of non-mycorrhizal barley was higher because of an impaired starch accumulation during grain filling. In mycorrhizal plants, leaf disease did not impair either the quantity or quality of grain yield. This improved compensation in mycorrhizal plants was related to maintained photosynthetic capacity and a delay in pathogen-induced senescence. Thus filling of long-term storage pools (fructans in internodes) and consequently reallocation of these reserves during grain filling was improved. The results suggest that higher availability of energy and material during grain formation, together with longer physiological activity, were the basis of yield maintenance and, therefore, expression of mycorrhiza-induced tolerance towards the pathogen.     

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

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