The effect of sowing date on adult plant resistance to Erysiphe graminis f.sp. avenae in oats

A technique developed for detecting small differences in mildew infection under glasshouse/laboratory conditions is described. A satisfactory correlation was found between results obtained by this method and infections in the field. Detailed examination of the levels of infection in cultivars Maldwyn and Milford showed that the upper and more recently developed leaves of Maldwyn had a significantly smaller area infected than those of Milford, although seedling leaves of both genotypes were very susceptible. This was most marked when plants were in the 6- to 7-leaf stage. Comparison of host plants sown on successive dates during the spring and early summer showed that with the acceleration of the rate of growth occurring in later-sown plants, resistance was expressed progressively earlier in the life of the plant. Newly expanded leaves which show very low level of infection at the 6- to 7-leaf stage become susceptible as they age, resistance being then expressed in the younger leaves.     

The preconditioning effect of day-length and light intensity on adult plant resistance to powdery mildew in oats

Oat genotypes with partial resistance to mildew exhibited a higher level of adult plant resistance (a.p.r.) when grown in 16 h compared with 8 h day-length. This increased resistance was expressed in the upper leaves, and to some extent in the lower, earlier-developed leaves. The expression of increased resistance coincided with floral initiation in Maldwyn, but preceded it in CC4761. The development of a.p.r. and floral initiation were thus not inherently linked, but conditions which accelerated the plants' ontogeny also caused earlier expression of resistance. For the most efficient testing of breeding material, plants should be grown at long day-length and high light intensity. When plants are grown at 16 h day-length and constant temperature of 15 ^oC, the leaf segments should be inoculated at the 6 to 8 leaf stage because normally susceptible genotypes showed marked resistance when inoculated later.     

The effect on grain yield of adult plant resistance to mildew in oats

Lines were selected from progeny of the crosses Maldwyn × Milford and Maldwyn × Sun II which consistently exhibited small and large amounts of mildew in similar situations. These are referred to as ‘low’ and ‘high’ mildew lines respectively. In small plot field trials conducted in 1971, 1972 and 1973 the low mildew lines produced substantially more grain each year compared with the high mildew lines in plots where mildew was allowed to develop normally. In plots which were kept comparatively free from infection by spraying with tridemorph, both low and high mildew lines gave similar yield. When the unsprayed plots were compared with the sprayed plots the low mildew lines invariably showed very small losses in grain yield and the high mildew lines usually very large losses. This same trend was evident also in the parent cultivars in 1972. Maldwyn, with only 7·9% mildew cover in the unsprayed plots due to its resistance, suffered a decrease of only 9·2% in grain yield compared with losses of 17·1 and 20·3% in the more susceptible cultivars Milford and Sun II. Certain lines and cultivars sustained heavy levels of mildew without corresponding loss of grain yield, i.e. showed low sensitivity to the disease. The importance of combining low sensitivity (or tolerance) with adult plant resistance to mildew is discussed.     

The Interaction of Humidity and Resistance Elicitors on Expression of Polygenic Resistance of Barley to Mildew

Elicitor-active glucan was applied to leaves of genotypes of barley expressing different levels of resistance to powdery mildew ( Erysiphe graminis f. sp. hordei )which was assessed by measuring mildew colonies per leaf or speed of papilla induction response to mildew challenge. The influence of humidity was also assessed by treating leaves with glucan, inoculating with mildew and immediately sealing the leaves in Perspex containers in an apparatus which enabled temperature, air speed and humidity to be controlled accurately. In water-treated controls low humidity alone reduced colonies per leaf, and at 60% r.h. genotype 7204 showed significantly less effect of humidity compared with other genotypes, although at 40% it responded as other genotypes. At low humidity glucan stimulated resistance for genotypes Proctor and 7204 but not for Golden Promise or 9855; the effect of humidity or elicitor alone was only significant on genotype 7204, Chitin also proved an effective elicitor, but at high and not at low humidity. Time of pre-treatment before mildew challenge, leaf stage, type of elicitor, concentration of elicitor, post-inoculation light regime, mildew isolate and barley genotype all had major influences on papilla response. Other factors were less important. Based on papilla formation, the barley cultivar Proctor was most and Golden Promise the least responsive to glucan as an elicitor. The potential for exploiting the genetic basis of inducible resistance is discussed.     

Inheritance of adult plant resistance to powdery mildew in swedes

A complete diallel set of crosses, including selfs, was produced from 11 inbred lines of swedes and assessed for adult plant resistance to powdery mildew in a replicated field trial. The plants were inoculated with powdery mildew spores on 23 July 1987. Traces of mildew were visible on 3 August. By 14 September the most susceptible inbred line had 95% of its leaf area infected, whereas the three most resistant lines derived from cvs Bangholm Dima (two lines) and Bangholm Magres (one line) had only 5%. Resistance was controlled by partially recessive genes. There was no evidence of maternal effects. The implications for breeding inbred and F₁ hybrid swede cultivars are discussed.     

The resistance to Oidium lycopersici conferred by ol-2 gene in tomato

The powdery mildew caused by Oidium lycopersici is one of the most destructive diseases in glass-house-grown tomato and is widespreading all over the world. A high level of resistance to O. lycopersici was found in an accession of Lycopersicon esculentum var. cerasiforme at the Department of Biology and Plant Pathology, University of Bari. The genetic analysis of F1, F2 and BC plants indicated that the resistance is conferred by a single recessive gene, designed as ol-2. Studies on the infection process of O. lycopersici on susceptible and ol-2 gene resistant tomatoes were carried out at 24 °C and 90 % relative humidity. Light microscope observations on conidia germination, formation of primary appressoria, elongation of hyphae and sporulation were made on artificially inoculated basal, intermediate and apical leaves. Inoculation was made by shaking mildewed tomato leaves over each test plant. Disease development were assessed by removing the fungal structure from the leaf surface with the ceroidin film technique and by direct observations of stained inoculated leaves. The rate of conidial germination and the appressoria formation was not affected by host genotype. Mycelia growth and sporulation on leaf surface of resistant plant was strongly restricted and influenced by the leaf age. The results indicated that the resistance in ol-2 tomato is postinfectional and is not associated with a hypersensitive response. This work was supported by the MURST and CNR (Paper no. 331)     

The influence of photoperiod on growth patterns of Erysiphe graminis f. sp. hordei

Detached seedling leaf segments of five spring barley genotypes were inoculated with an isolate of barley mildew to which they possessed different levels of resistance. Segments of each host genotype were then incubated under either continuous light or treatments with 2, 4, 8 or 16 h darkness per 24 h cycle. Macroscopic observation showed that the latent period of infection was reduced slightly in treatments with at least 4 h darkness/24 h. Yellowing of detached segments occurred fastest under continuous light and slowest under a 16 h dark/8 h light cycle. Microscope observation of segments fixed 4·5 days after inoculation showed that as the length of the dark period increased, so the number of haustoria formed per colony also increased. This increase in haustorial production appeared to be associated with an accentuation in the synchrony of production of the secondary and tertiary haustorial generations. Varietal differences in susceptibility were also more marked in segments incubated under short days. It is concluded that under the conditions of temperature and light intensity used, a 16 h dark/8 h light system is most desirable for quantifying mildew resistance because it allows confident identification of distinct haustorial generations, accentuates differences in varietal susceptibility and delays chlorophyll degradation in detached barley leaves infected with mildew.     

Components of Partial Disease Resistance in Wheat Detected in a Detached Leaf Assay Inoculated with Microdochium majus using First, Second and Third Expanding Seedling Leaves

The use of first, second and third expanding seedling leaves of wheat (L1, L2 and L3 respectively), inoculated with conidial suspensions of Microdochium majus (syn. Microdochium nivale var. majus) in a detached leaf assay, for detecting components of partial disease resistance (PDR) was investigated across a range of wheat cultivars. Incubation periods (period from inoculation to first appearance of symptoms; a dull grey–green water‐soaked lesion) and latent periods (period from inoculation to the first appearance of sporodochia) were longest and lesions smallest on L3. The expression of PDR components on L2 was intermediate to those on L1 and L3. The longer latent periods on L3 typically occurred after leaf senescence contrasting with latent periods on L1 and L2 where sporulation most frequently occurred on relatively green leaf tissue. Cultivar differences in the first appearance of symptoms, incubation period, which occurred before any leaf senescence was observed, correlated significantly across all leaf positions. Similarly cultivar differences in latent period were correlated for L1 and L2. However, latent periods on L3, which were the least consistent between cultivars across experiments, were not correlated with those of L1 or L2 in any experiment. The results indicate that due to the delay in sporulation until after leaf senescence, observations on latent period in L3 are less representative of what occurs in the whole plant where infection of living tissue is of greatest interest. This work indicates that the selection of the first or second expanding leaf of wheat is optimal for the use in the detached leaf assay using M. majus for studying components of PDR.     

Sensibilité du Melon àSphaerotheca fuliginea en Fonction de L'Etage Foliaire

Effect of leaf position on the susceptibility of melon plants to artificial infection with powdery mildew, Sphaerotheca fuliginea The leaf position of melon plants seems to play a role on their susceptibility when they are artificially infected with powdery mildew Sphaerotheca fuliginea. The cotyledons are generally very susceptible, while the first leaf relatively resistant; the susceptibility again continues up to the 4th–5th leaf (but less susceptible than the cotyledons) and then after it decreases; these results can be obtained on plants in greenhouse or on detached leaves in Petri dishes. From this observation, we think that the screening of melon genotypes for resistance to powdery mildew can be evaluated neither on the cotyledons which are very susceptible nor on the first leaf which is resistant, but on the third leaf which is moderately susceptible. In fact, there is a good correlation between the reaction of the third leaf and the resistance or susceptibility of genotypes.     

In vivo localization at the cellular level of stilbene fluorescence induced by Plasmopara viticola in grapevine leaves

Accurate localization of phytoalexins is a key for better understanding their role. This work aims to localize stilbenes, the main phytoalexins of grapevine. The cellular localization of stilbene fluorescence induced by Plasmopara viticola, the agent of downy mildew, was determined in grapevine leaves of very susceptible, susceptible, and partially resistant genotypes during infection. Laser scanning confocal microscopy and microspectrofluorimetry were used to acquire UV-excited autofluorescence three-dimensional images and spectra of grapevine leaves 5-6 days after inoculation. This noninvasive technique of investigation in vivo was completed with in vitro spectrofluorimetric studies on pure stilbenes as their fluorescence is largely affected by the physicochemical environment in various leaf compartments. Viscosity was the major physicochemical factor influencing stilbene fluorescence intensity, modifying fluorescence yield by more than two orders of magnitude. Striking differences in the localization of stilbene fluorescence induced by P. viticola were observed between the different genotypes. All inoculated genotypes displayed stilbene fluorescence in cell walls of guard cells and periclinal cell walls of epidermal cells. Higher fluorescence intensity was observed in guard-cell walls than in any other compartment due to increased local viscosity. In addition stilbene fluorescence was found in epidermal cell vacuoles of the susceptible genotype and in the infected spongy parenchyma of the partially resistant genotype. The very susceptible genotype was devoid of fluorescence both in the epidermal vacuoles and the mesophyll. This strongly suggests that the resistance of grapevine leaves to P. viticola is correlated with the pattern of localization of induced stilbenes in host tissues.     

Components of partial resistance as criteria for identifying resistance

The components of partial resistance, incubation period, lesion area, latent period and sporulation were recorded on plants of six winter and two spring wheat cultivars which had been artificially inoculated with Septoria nodorum spores. Incubation period gave a guide as to how the cultivars would respond in the field to Septoria nodorum but statistical analysis showed that it could not be used alone to predict accurately the resistance of each cultivar to the pathogen. Average sporulation, however, could be used with more confidence for predicting the field resistance of the cultivars. From a regression analysis of NIAB rating versus incubation period, lesion area, latent period and sporulation, an equation was devised to obtain resistance indices for each cultivar. These resistance indices clearly reflected the NIAB ratings for the cultivars. It would therefore appear that resistance indices could be used as a pre-field evaluation method for identifying resistance to Septoria nodorum and thus be a valuable technique in breeding programmes.     

Canopy development of a determinate and an indeterminate cultivar of Vicia faba L. under contrasting plant distributions and densities

Field experiments were conducted in 1987 and 1988 to quantify differences in canopy formation between an indeterminate and a determinate genotype of Vicia faba L., grown at two plant densities and three spatial distributions. The number of stems per unit area produced by determinate plants was related to the growth rate before flowering. Leaf production per stem per unit of thermal time was similar in both plant types, but twice as many leaves per stem were produced by the indeterminate cultivar. The indeterminate cultivar produced fewer and smaller leaves in the warmer and drier weather of 1988 than in 1987. The determinate genotype produced similar sizes and numbers of leaves in both years, but fewer tillers developed in 1988 than in 1987. Accordingly, leaf mass per unit ground area was greater in 1987 than in 1988 in both genotypes. Except during early flowering, relationships between leaf mass and leaf area were constant, with higher specific leaf areas in the determinate than the indeterminate genotype. Shoot dry matter partitioning into leaves was identical in both years for indeterminate plants, but differed in determinate ones.
It is concluded that canopy development is regulated through individual leaf weight and leaf number per stem in non-tillering indeterminate, and by stem numbers per unit area in tillering determinate plants.     

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.     

A Method to Measure Aggressiveness of Plasmopara halstedii (Sunflower Downy Mildew)

For the first time, a method was used to measure aggressiveness of two Plasmopara halstedii races (100 and 710), the parasite causing sunflower downy mildew. Two sunflower lines showing different levels of quantitative resistance were used to measure two aggressiveness criteria: latent period and sporulation density. A strain of race 100 had a shorter latent period and greater sporulation density than a strain of race 710. The sunflower inbred line BT, rather susceptible in the field, presented a greater sporulation density and a shorter latent period than another inbred line FU, which shows greater resistance in the field. These results indicated that race 100 was more aggressive than race 710. The behaviour in the field of the two inbred lines was confirmed in the laboratory observations.     

Effects of Alteria alternata f.sp. lycopersici toxins at different levels of tomato plant cell development

Since the host-specific toxins of Alternaria alternata f. sp. lycopersici play an important role in pathogenesis, they potentially could be applied as selective agents in in vitro selection at the cellular level for disease resistance. Prerequisite for this is that sensitivity to the Alternaria alternata f.sp. lycopersici pathotoxins is manifest at the cellular level. To gain insight into cellular effects of AAL-toxins and into the mechanisms of plant insensitivity to AAL-toxins, effects of AAL-toxins on leaves, leaf discs, roots, calli, suspension cells, minicalli and protoplasts of susceptible and resistant tomato genotypes were studied. In leaves of susceptible genotypes, toxins cause severe necrosis, while in leaves of resistant genotypes necrosis was never observed. Inhibition effects of toxins were observed at all other levels in susceptible and resistant genotypes: toxins inhibited shoot induction on leaf discs, root growth and growth of calli, suspension cells and protoplasts. This indicates a cellular site for AAL-toxins. Differences in sensitivity to AAL-toxins between susceptible and resistant genotypes were observed in leaves and roots, but were not observed during shoot induction on leaf discs, in calli, suspension cells and protoplasts. However, differences in sensitivity to AAL-toxins in roots were at least 20 times less than in leaves. Therefore insensitivity seems related to a higher level of tomato plant differentiation and is most pronounced in leaves.     

Comparison of Different Inoculation Techniques to Screen Resistance of Pea Lines to Downy Mildew

Pea lines with partial resistance to downy mildew were tested with different inoculation techniques. Lines with superior resistance to leaf and pod infection in the field also show superior resistance to artificial inoculation of germinating seeds or seedlings at different ages in greenhouse experiments. Increased resistance of older seedlings was found for all genotypes including the susceptible control. Juvenile leaves at the top of the seedlings plants at time of inoculation, developed the strongest symptoms. Selection for partial resistance can probably be carried out in the greenhouse to improve resistance of adult plants in field.     

Expression analysis of defence-related genes in grapevine leaves after inoculation with a host and a non-host pathogen.

The expression of PR protein encoding genes and genes involved in the phenylpropanoid metabolism was analysed on grapevine leaves of susceptible and resistant cvs. in response to inoculation with the host-pathogen Plasmopara viticola and the non-host pathogen Pseudoperonospora cubensis, the downy mildew pathogen of cucumber. These experiments were conducted to elucidate whether or not grapevine plants susceptible to downy mildew exhibit an identical defence response after inoculation with the non-host pathogen. Expression analysis of defence-related genes revealed marked differences between the susceptible cultivar "Riesling" (Vitis vinifera) and the resistant cultivar "Gloire de Montpellier" (Vitis riparia). Whereas some genes seem to be expressed constitutively in "Gloire" or induced after an inoculation with both pathogens, expression of defence-related genes in Riesling was influenced mainly after inoculation with the non-host pathogen: PR-2, PR-3, PR-4, a PGIP gene, and especially genes encoding enzymes involved in anthocyanin biosynthesis (DFR, F3H, LDOX) were affected. Therefore, the occurrence of the respective products (flavans and other phenolics) in inoculated leaves was investigated with appropriate histological staining techniques. These stainings revealed a production of catechins and related phenolic compounds within the first 48 hai (hours after inoculation) with Ps. cubensis but not with P. viticola in Riesling, whereas in Gloire no further production was seen, which may be due to the high content of polyphenolics as observed in control leaves. In addition to the staining procedures, sporulation intensity was monitored on leaf discs. Pretreatments of leaf discs with Ps. cubensis led to a reduced browning reaction (as a result of a hypersensitive reaction) in Gloire and significantly reduced the intensity of sporulation in Riesling after a subsequent inoculation with P. viticola.     

Lack of trade-off between direct and indirect defence against grape powdery mildew in riverbank grape

Abstract.  1. The hypothesis that plants that invest more in an indirect defence will invest less in direct mechanisms was tested using genotypes of the riverbank grape Vitis riparia that varied in expression of acarodomatia, tufts of non-glandular trichomes on underside of leaves that are frequently occupied by mycophagous mites. Mycophagous mites in this system have been shown to protect vines from grape powdery mildew caused by the fungus Uncinula necator .
2. Multiple cane cuttings from 24 genotypes of V. riparia , a species native to North America, were obtained from the USDA grape genetics repository and planted out in a common garden.
3. Genotypes varied in size of acarodomatia by threefold and varied in density of mycophagous mites by over 10-fold. Genotypes originating from the western part of the distribution had smaller acarodomatia and fewer mycophagous mites than genotypes from the eastern part of the distribution. There was a positive genetic correlation between acarodomatia size and mite density.
4. In a separate experiment, where vines were kept free of mycophagous mites, genotypes showed 10-fold variation in percentage leaf surface colonised by mildew. There was, however, no relationship between mildew on leaves and size of acarodomatia. Mildew did not vary depending on origin of genotype, although female vines tended to have greater levels than males.
5. No evidence was found for a trade-off between investment in an indirect defence mediated through acarodomatia and investment in direct defence against grape powdery mildew. Mite/plant defence mutualisms are contrasted with ant/plant defence mutualisms where there is better evidence for trade-offs under field conditions.     

High levels of auto-infection in plant pathogens favour short latent periods: a theoretical approach

Auto-infection (infection arising from inoculum produced on the same host unit) is common in polycyclic plant pathogens, but often neglected in experimental and theoretical studies, which focus instead on infection of new hosts (allo-infection). Here we test the hypothesis that high auto-infection, as observed for leaf infecting fungal pathogens, could select for short latent periods. An individual-based simulation model keeps track of lesions, resulting from the spread of spores, between and within individual leaves. Linked to a trade-off between latent period and spore production capacity, as observed for Puccinia triticina on wheat, the adaptation of the latent period is analysed for different levels of auto-infection using the methods of pairwise invasibility plots. Results suggest that increased auto-infection selects for reduced latent periods. A reduction in leaf longevity also selects for reduced latent periods, which is most obvious for a relatively low ratio of auto- to allo-infection. This study is the first to consider the effect of auto-infection on the evolution of pathogen life history traits. The fact that auto-infection could drastically reduce pathogen latent periods highlights the need for more research in this area.     

Morphophysiological and biochemical attributes influence intra-genotypic preference of shoot fly [Atherigona soccata (Rondani)] among sorghum genotypes

Shoot fly [Atherigona soccata (Rondani)] is a destructive pest of sorghum at the seedling stage and causes huge losses to grain yield and green fodder. The host-plant resistance mechanism is the best approach to reduce the attack of insects in plants. The damage parameters, morphophysiological traits, and biochemical metabolites had been investigated in the leaves and stem of contrasting sorghum genotypes, viz., resistant (IS18551, ICSV705, ICSV700), moderately resistant (PSC-4), and susceptible (SWARNA and SL-44) at 15 and 21 days after emergence (DAE) against shoot fly infestation. The resistant genotypes recorded lowest shoot fly oviposition and incidence (0.3–0.7 eggs plant⁻¹ and 10–15%) than the susceptible genotypes (2.4–3.0 eggs plant⁻¹ and 70–80%), respectively. The susceptible genotype SWARNA recorded 50% and 80% higher deadheart formation than the resistant genotype IS18551 at 15 and 21 DAE, respectively. Resistant genotypes exhibited higher trichome density at adaxial and abaxial part of leaf (118–145 and 106–131) with pink colored leaf sheath (scale 1.50–3.25), glossy leaves (scale1.00–1.25), and lower leaf surface wetness (scale1.25–2.00) compared with susceptible genotype with 49.3–73.3 and 25.3–64.0, scale 2.50–4.00, scale 2.75–3.50, and scale 3.25–4.25 for the respective parameters. Another defense response of sorghum toward the insect attack was modulation of plant metabolism. The infested genotypes responded to insect attack by upregulation of total soluble sugar, total phenol, prussic acid, and chlorophyll content by 1.2–2.1-fold, 1.5–2.0-fold, 1.2–1.3-fold, and 1.2–3.9-fold with more induction in susceptible genotypes at 21 DAE. On the whole, the present study indicates that morphophysiological and biochemical attributes contribute toward the resistance mechanism in sorghum against shoot fly infestation.