Carbon Dioxide Exchange in an Incompatible Barley/Powdery Mildew Combination

CO₂ exchanges in intact barley leaves were studied by infra red gas analysis after inoculation with an incompatible race of powdery mildew. Net photosynthesis was inhibited, and dark and photorespiration were stimulated, by inoculation. The increase in respiratory processes was approximately equal to the change in net photosynthesis indicating that CO₂ fixation, “gross photosynthesis”, was unaffected by inoculation. It is concluded that changes in net photosynthesis may be the primary cause of the reduced quantity and quality of grain in incompatible barley/mildew combinations.     

On the Enhanced Callose Deposition in Barley with ml-o Powdery Mildew Resistance Genes

Carborundum treatment of barley leaves induced a callose deposition which was detected as diffuse blotches in the epidermal cells of susceptible barleys and as deeply stained tracks along the scratches in barleys with the ml-o powdery mildew resistance gene. Subsequent inoculation with powdery mildew resulted in appositions that enlarged inversely to their size in the respective varieties when inoculated without carborundum treatment. Aphids sucking the leaves resulted in rows of callose containing spots along the anticlinal cell walls. The spots were larger in the ml-o mutant than in the mother variety. Callose was deposited in connection with the pleiotropic necrotic spotting in barleys with the ml-o gene. Modification of the necrotic spotting by crossing the ml-o gene into other gene backgrounds did not result in any change in the size of appositions upon inoculation with powdery mildew. Callose deposition was never observed in cells subsidiary to the guard cells, and the absence of callose in such cells is suggested to be the reason for the sporadic occurrence of powdery mildew colonies on barleys with the ml-o gene.     

Genetics of Powdery Mildew Resistance in Barley

A comprehensive, critical review on the present knowledge regarding the genetics of resistance of barley to the powdery mildew fungus is presented. The review deals with six kinds of resistance: Race-specific resistance; Mlo resistance; partial resistance; induced resistance; passive resistance; and non-host resistance. Most of the sections are subdivided into: phenotype of the interaction; resistance mechanisms; and genetics. A distinction is made between three groups of genes involved in the defense of plants to diseases: those that serve exclusively to mediate resistance; those that are mobilized to strengthen the plants' defense; and those that serve exclusively functions other than disease defense, but may bring about resistance. The more than 200 gene symbols assigned to race-specific mildew resistance genes over time are summarized and revised to 85 symbols that may be considered valid.     

Powdery Mildew Resistance in Barley Landraces from Morocco

Nineteen barley landraces collected from Morocco were screened for resistance to powdery mildew. The landraces originated from the collection at the Polish Gene Bank, IHAR Radzików, Poland. The fifteen landraces tested showed powdery mildew resistance reactions and 35 single plant lines were selected. Twenty-one of these lines were tested in the seedling stage with 30, four lines with 17 and another 10 lines with 23 differential isolates of powdery mildew, respectively. The isolates were chosen according to their virulence spectra observed on the Pallas isolines differential set. Nine lines (E 1029-1-1, E 1042-2-2, E 1050-1-1, E 1054-5-1, E 1056-2-5, E 1056-3-1, E 1061-1-1, E 1061-1-3 and E 1067-1-2) which originated from seven landraces showed resistance to all prevalent European powdery mildew virulence genes. The most frequent score was 2 and 16 lines showed this reaction for inoculation with most isolates used. The distribution of reaction type indicated that about 77% of all reaction types observed were classified as powdery mildew resistance (scores 0, 1 and 2). In all lines the presence of unknown genes alone or in combinations with specific ones was postulated. Four different resistance alleles ( Mlat , Mla6 , Mla14 and Mla12 ) were postulated to be present in 10 tested lines alone or in combination. Alleles Mlat , Mla6 and Mla14 were postulated to be present in four and Mla12 in two tested lines, respectively. The value of barley landraces for diversification of resistance genes for powdery mildew is discussed.     

Effect of Nitrogen and Powdery Mildew on the Yield Formation of Two Winter Barley Cultivars1

In field experiments the effect of powdery mildew on the yield formation of two barley cultivars ‘Catinka’ (partially resistant) and ‘Tapir’ (susceptible) was investigated in relation to three levels of nitrogen fertilization (40, 90, 150 kg N/ha). Nitrogen increased especially the single ear yield of cv.‘Catinka’, whereas cv.‘Tapir’ showed symptoms of N oversupply. Infection density of Erysiphe graminis hordei also increased with increased nitrogen supply, although it decreased transiently, at the highest N level for cv.‘Tapir’. In general, cv.‘Catinka’ was less tolerant to infections with E. graminis than cv.‘Tapir’, but yield response of both cultivars to powdery mildew (17, 11 and 8 % and 12, 16, and 12 % yield losses for cv.,‘Catinka’ and ‘Tapir’. respectively) showed cultivar × nitrogen interrelationships. Starch and protein contents of grains indicated differences in the intensity of detrimental effects caused by powdery mildew limiting the formation of carbohydrates and/or the translocation processes. Regression analysis demonstrated a closer correlation of yield losses with the photosynthetically active leaf area (r =+0.96 and +0.93 for ‘Catinka’ and ‘Tapir’) than with disease intensity (r =−0.32 and −0.85). Theinte,rpretation of disease/yield loss relations and the detrimental effect of powdery mildew are discussed with reference to plant resistance, remaining green leaf area, the availability of nutrients, to ‘sink and source’ conditions and to the occurrence of,additional environmental stresses.     

Quantitative Differences in Powdery Mildew Resistance Among Spring Barley Cultivars

Quantitative powdery mildew resistance in compatible host-pathogen-combinations was measured by the number of pastules/cm² leaf area. Spring barley cultivar ‘Proctor’ was significantly less infected than ‘Golden Promise”. Using these two cultivars (having no effective major resistance gene) as controls, MO- and AR-resistant cultivars were inoculated with virulent mildew isolates. ‘Mona”, ‘Grit’ and ‘Nudinka’ had a higher or, at least, the same level of quantitative resistance as ‘Proctor”. None of the remaining cultivars showed the high susceptibility expressed by ‘Golden Promise”. Ranking of host genotypes was nearly constant while that of mildew isolates varied considerably. Only a small portion of the observed variance was due to interaction between host cultivars and pathogen isolates. ‘Triesdorfer Diva’ gave a resistant infection type after inoculation with different AR-virulent isolates, indicating that this cultivar has major resistance other than that conditioned by gene Ml-a12.     

The Ability of Barley Powdery Mildew to Grow in vitro

A technique was developed for the in vitro culture of Blumeria graminis f. sp. hordei , an obligate biotrophic pathogen of barley. Optimal growth occurred at pH 5.6 on a medium containing 39 gl^–1 potato dextrose agar, 40 gl^–1 shredded fresh barley leaves, 20 gl^–1 sucrose, 13 mgl^–1 kanamycin and 80 mgl^–1 benzimidazole. At 20°C (90% relative humidity), conidia germinated 48 h after inoculation, producing an average colony diameter of 1 cm after 10 days. However, numerous colonies were present on the medium after 15 days. Light microscopy showed that there was a positive relationship between the amount of leaf in the medium and fungus growth. The fungus retained its virulence during 60 days of storage in vitro , and was able to infect barley. This is a useful and novel technique that could be beneficial in barley pathology breeding programs.     

Distribution of the Products of Photosynthesis between Powdery Mildew and Barley

The photosynthetic assimilation of ¹⁴CO₂ has been studied in healthy and mildew-infected barley. The parasite was separated from the host by removing the mycelium with a camel's hair brush. The ethanol soluble metabolites of the parasite, infected host and healthy host were extracted, separated by paper chromatography and individually identified. From this work it appears that there is a rapid movement of label from host to parasite mainly in the form of sucrose which is then quickly metabolized into many compounds. The majority is converted into mannitol, and lesser amounts are converted into trehalose, arabitol, aspartic acid, and glutamic acid. In conidia the major carbon reserve is arabitol instead of mannitol, with lesser amounts of trehalose and mannitol.

Photosynthetic uptake of ¹⁴CO₂ by the complex decreases steadily after inoculation as compared with healthy leaves. However, the ethanol soluble metabolites of the infected host tissue differ only slightly from those of healthy host tissue. The major differences are a slight decrease in the amount of sucrose and increases in malic acid and serine.

     

Hordein Variation and Reaction to Powdery Mildew in Composite Cross XLII of Barley

A bulk hybrid population, CCXLII was investigated for hordein variation and reaction to powdery mildew. The results indicated that the population in F₄ was genetically variable and contained an appreciable proportion of heterozygotes. Evidence was found for differential viability within families. This was possibly the result of a high segregation load. The pattern of genetic variation suggests that although the population could be a useful source of breeding material for the selection of new lines, it may be risky as a method of conservation of germplasm.     

Blufensin1 Negatively Impacts Basal Defense in Response to Barley Powdery Mildew

Plants have evolved complex regulatory mechanisms to control the defense response against microbial attack. Both temporal and spatial gene expression are tightly regulated in response to pathogen ingress, modulating both positive and negative control of defense. BLUFENSIN1 (BLN1), a small peptide belonging to a novel family of proteins in barley (Hordeum vulgare), is highly induced by attack from the obligate biotrophic fungus Blumeria graminis f. sp. hordei (Bgh), casual agent of powdery mildew disease. Computational interrogation of the Bln1 gene family determined that members reside solely in the BEP clade of the Poaceae family, specifically, barley, rice (Oryza sativa), and wheat (Triticum aestivum). Barley stripe mosaic virus-induced gene silencing of Bln1 enhanced plant resistance in compatible interactions, regardless of the presence or absence of functional Mla coiled-coil, nucleotide-binding site, Leu-rich repeat alleles, indicating that BLN1 can function in an R-gene-independent manner. Likewise, transient overexpression of Bln1 significantly increased accessibility toward virulent Bgh. Moreover, silencing in plants harboring the Mlo susceptibility factor decreased accessibility to Bgh, suggesting that BLN1 functions in parallel with or upstream of MLO to modulate penetration resistance. Collectively, these data suggest that the grass-specific Bln1 negatively impacts basal defense against Bgh.     

Quantitative Analysis of the Early Powdery Mildew Infection Stages on Resistant Barley Genotypes

A classification system was developed, that allowed quantification of the leaf surface development of the barley powdery mildew fungus on barley. An experiment with Manchuria and Pallas as susceptible controls and 4 resistance gene each represented by three lines with different gene backgrounds showed two types of gene background effects. First a general effect comprised of different distributions of the elongating secondary hyphae (ESH) in the stomatal and interstomatal region between Manchuria and, Pallas, and a higher number of lobes per appressorium on Pallas than on Manchuria. These effects also applied to isolines of Manchuria and Pallas possessing each of the four genes investigated. Second a specific effect was noticed on the ESH frequency on Ml-k lines. The ESH frequency varied significantly at the 5% level between the highest and lowest value. An experiment with Pallas as susceptible control and Pallas isolines with 11 different resistance genes, showed that powdery mildew development was unaffected by host genotype until after the formation of an appressorium. The first host effect observed was on the number of lobes on the appressorium, which reflects the number of penetration attempts. This number increased as the degree of resistance increased, i.e. the ESH frequency decreased. The penetration stage also invariably proved to be the limiting stage, where the largest proportion of fungal propagules was stopped.     

Isozyme Variation and Genetic Diversity of the European Barley Powdery Mildew Population

Isozyme and virulence analyses of Erysiphe graminis bordei were performed with samples collected from different sites from nearly all over Europe. Isozymes and unspecific proteins extracted from conidia were separated by starch gel electrophoresis and isoelectric focusing, respectively, and the resulting isozyme banding patterns were compared with the corresponding virulence data. One isozyme phenotype prevailed in all samples. Only 7.9% of 280 isolates showed divergent banding patterns. Expected frequencies of isolates with divergent banding patterns were calculated for each subsample. In the Italian subsample, isolates with divergent banding patterns were significantly more frequent than expected. At the same time, isolates from Italy had significantly fewer virulence factors than those from N.W. Europe, indicating weaker selection by host resistance genes. It is suggested that isozyme uniformity in the homogeneous north-western European barley powdery mildew population has arisen from strong selection pressures for specific virulence genes. The direction of this selection, acting upon a mainly asexually reproducing population, has changed over space and time due to the introduction of new resistance genes, forcing local populations through bottlenecks. This may have led to random loss of genetic variation (genetic drift) in the barley powdery mildew gene pool.