Strategies for control of eyespot (Pseudocercosporella herpotrichoides) in UK winter wheat and winter barley

This review considers strategies for control of eyespot (Pseudocercosporella herpotrichoides) in winter wheat and winter barley in the UK. Varietal resistance to eyespot may take the form of direct resistance to the growth of the pathogen in the stem base or of indirect tolerance to eyespot through resistance to lodging. The French variety Cappelle-Desprez has been a source of resistance for most UK wheat varieties and a new source of resistance in wild goat grass has been discovered recently. Use of fungicides for control of eyespot increased rapidly after the introduction of the highly effective MBC fungicides in the 1970s, but has decreased recently because UK populations of P. herpotrichoides are now predominantly resistant to MBC fungicides and alternative fungicides are more expensive. Historically, cultural methods, especially crop rotation, have been important in the control of eyespot. The importance of cultural control of eyespot is now increasing because more non-cereal break crops are being grown and fungicide use is declining.     

Predisposing effects of Eyespot (Pseudocercosporella herpotrichoides) on Septoria nodorum infection of winter wheat

All isolates of PeniciUium simplicissimum, P. verrucosum var. cyclopium, P. brevicompactum, P. multicolor, P. oxalicum, P. paxilli, Botrytis cinerea, and of a Gliocladium sp. obtained from necrotic virus-tested narcissus twin-scales previously dtoped in benomyl were tolerant to 1000 μg/ml of this fungicide in agar. Every necrotic twin-scale examined was infected with at least one of these species. The first two species were the most frequently isolated and a similar range of species infected different narcissus clones or different cultivars. It is argued that the use of benomyl during the twin-scaling programme should be discontinued as soon as a suitable alternative fungicide is found.     

Sensitivity to prochloraz in populations of the eyespot fungus, Pseudocercosporella herpotrichoides, in relation to fungicide treatments and their efficacy in continuous winter wheat

Eyespot was assessed and grain yields determined in the eighth and ninth years (1992 and 1993) of a field experiment in which the fungicides carbendazim and prochloraz were applied, separately or in mixtures, to plots of successive crops of winter wheat. Populations of the eyespot fungus, Pseudocercosporella herpotrichoides, were characterised by the proportions of cultures grown on agar from infected stems that were W-type or R-type, or were carbendazim-resistant or carbendazim-sensitive. Sensitivity to prochloraz in agar was determined for isolates from populations sampled in 1992 using restricted maximum likelihood analysis of EC50s (concentrations needed to decrease colony growth by 50%), to deal with the unbalanced data, and comparisons were made by Wald statistics. Control by prochloraz was maintained but in 1992, as in some previous years, its application with carbendazim was more effective than its application alone. Selection by prochloraz for strains sensitive to carbendazim continued to occur and may have contributed to its sometimes relatively poorer performance in the absence of carbendazim. R-type isolates taken from prochloraz-treated plots, with or without carbendazim, in 1992 were less sensitive to prochloraz than were isolates from plots untreated with prochloraz. There was evidence of a greater range of sensitivities to prochloraz in R-type than in W-type isolates (although some uncertainty results from the small numbers of W-type isolates obtained from some treatments), which would explain the selection of the R-type by prochloraz. The significance of these findings to modern wheat growing practices is considered.     

The effect of temperature on eyespot (Cercosporella herpotrichoides) in wheat seedlings

When seedlings of the winter wheat cultivars Cappelle-Desprez, Champlein and Hybrid 46 were artificially infected with mycelial inoculum of Cercosporella herpotrichoides Fron, the number of leaf sheaths penetrated by the pathogen increased with temperature over the range 6-18^oC. Rate of growth of the pathogen on agar also increased with temperature over this range. Temperature had little effect on differences between cultivars in number of leaf sheaths penetrated.     

Effects of eyespot on the yield of winter wheat

In two experiments, on Julia and Zephyr spring barley, evidence was obtained that the development of powdery mildew in sprayed plots depended very much on their proximity to unsprayed crop. The effective period of control achieved with one spray was greater where plots were well separated from unsprayed crop than where they were surrounded by or very close to unsprayed crop. There is probably a risk that small plot experiments may not only underestimate the potential effectiveness of fungicides in agricultural practice, but also give misleading information on optimal spray timing.     

Production and Regeneration of Protoplasts from Pseudocercosporella herpotrichoides (Fron) Deighton

This paper describes the development of a system for protoplast isolation from this pathogen using the commercially available, hydrolytic enzyme preparations Rhozyme HP150, Driselase and Cellulase CP. An analysis of the active components of these enzymes believed to be involved in protoplast release has been made. Factors affecting protoplast release including mycelial age, enzyme concentration and choice of osmotic stabilizer have been optimised, and conditions for high frequency protoplast regener, ation have been determined. The possible significance of the interaction of the several components of each enzyme is discussed. The developments described in this paper can now be exploited in genetic studies in this imperfect fungus.     

The effect of eyespot (Cercosporella herpotrichoides Fron.) on wheat and the influence of nitrogen on the disease

The effect of eyespot throughout the season on wheat receiving different amounts of nitrogen was studied in pot experiments. Plants inoculated in December showed chlorosis of outer leaves in February. Among plants with high nitrogen, eyespot killed 11%, caused straggling of 31% and whiteheads in 14% of the surviving ear-bearing straws, reduced yield of straw by 8% and of grain by 16%. The loss in straw yield was due to reduction in plant number, that of grain was about half due to reduction in number of ears and half to production of lighter grains. Among plants with low nitrogen the disease killed 23% of the plants, caused straggling of 86% and whiteheads in 18% of the surviving ear-bearing straws, and reduced yield of straw by 23% and of grain by 44%. The loss in straw yield was due to death of plants, that of grain was about two-thirds due to the production of fewer ears and one-third to that of lighter grains. In the high-nitrogen series the number of shoots at the time of maximum tillering was reduced by 29%; in the low-nitrogen series the disease caused reduction in height, a very uneven crop, delay in ear and anther emergence, and an increase in tail corn from 4% in the controls to 30% in the inoculated plants.
All inoculated plants became infected, but those receiving high nitrogen had only 49% of the ear-bearing straws with severe lesions at harvest, while those receiving low nitrogen had 86%. The larger number of tillers produced when nitrogen was applied may have enabled the less severely diseased shoots to survive and bear ears while the most severely infected died.     

Differentiation and Diagnosis of Pseudocercosporella herpotrichoides (Fron) Deighton with Genomic DNA Probes

Repetitive genomic clones were used to differentiate between varieties within the species Pseudocercosporella herpotrichoides . From 21 clones tested 13 revealed restriction fragment length polymorphisms among isolates. Cluster analysis was performed based on these data. Differentiation of isolates according to growth speed corresponds better to genetic information than differentiation on the basis of spore morphology. Pathogen-specific probes can be used in early diagnosis of the pathogen in infected plant material. Furthermore polymorphic pathogen-specific probes allow varieties to be differentiated directly in infected plant material without isolation and culture of the fungus.     

Detection of Pseudocercosporella herpotrichoides (Fron) Deighton in Wheat by Indirect ELISA

An indirect ELISA for quantitative detection of P. herpotrichoides in fections in wheat is presented. All tested isolates of the virulent varieties P. herpotrichoides var. herpotrichoides, P. h. var. acuformis or the W- and R-type react on a high level in the test, while the less virulent P. anguioides is assessed only with 40% and the avirulent P. aestiva with 20% of the homologous reaction. No crossreactions occur with extracts of 11 other species of in, vitro cultivated fungi nor with plant material infected with other pathogens. The infection profile throughout the leaf-sheaths was clearly reflected by ELISA. The examination of 24 stembase samples from the field showed that the values assessed by ELISA correlate well also with the disease indices of naturally infected plant material.     

Pathogenicity to Wheat of Isolates of Pseudocercosporella herpotrichoides (Fron) Deighton from Five Graminaceous Hosts

Eleven isolates of Pseudocercosporella herpotrichoides were obtained from field material of the Gramineae Agropyron repens, Alopecurus myosuroides, Apera spica-venti, Hordeum vulgare and Triticum aestivum. Four of these isolates belonged to the variety acuformis and 7 isolates to the variety herpotrichoides of the fungus. In a greenhouse experiment, winter wheat was inoculated with these isolates. All isolates were pathogenic on wheat. They differed in virulence, but these differences could not be related to the nature of their original hosts nor to their classification as variety acuformis or herpotrichoides.     

Effect of previous crops on the incidence of eyespot on winter wheat

Surveys of winter wheat from 1939 to 1946 show that eyespot (Cercosporella herpotrichoides Fron.) occurs throughout Britain and that its incidence depends largely on previous cropping and on weather. Examination of 551 crops on land whose cropping for the previous 4 years was known showed that the incidence rose steadily with increasing numbers of preceding wheat and barley crops: where neither crop had been taken for 4 years the proportion of crops with more than 70% infected straws was 2%, rising to 45% where three or four such crops had been taken and the average straws infected rose from 6 to 55%.
The percentage infection to be expected in various groups of crops was calculated from previous cropping; it was compared with the actual infection and so used to assess the importance of other factors in determining the incidence of eyespot. High spring rainfall, early sowing and a dense plant increased incidence and low spring rainfall, late sowing and a thin plant reduced it.
Eyespot was not usually severe on newly ploughed grassland until the third or fourth crop of wheat, but under very wet conditions it was sometimes severe in the second crop.
Oats is much less susceptible than wheat or barley, but some crops were found with a third of their straws infected.
A brief survey of winter wheat in Holland suggested possible causes for the rise and fall of eyespot in recent years and for its present lower incidence there as compared with East Anglia.     

Long-term performance of fungicides applied to control eyespot in winter wheat

Fungicide treatments, repeated twice yearly at timings suitable for controlling eyespot, to plots of continuous winter wheat over 17 yr (except for a year in “set‐aside”), resulted in altered population structures of the eyespot pathogens (Tapesia spp.) and performance of the fungicides. Carbendazim failed to control eyespot after 3 yr of the experiment because of rapid selection for resistance. Carbendazim often increased disease compared with that in untreated plots. This may have resulted from control of unknown antagonistic fungi or, possibly, from selection for increased fitness in carbendazim ‐resistant strains. In the final years of the experiment, prochloraz, applied with or without carbendazim, became ineffective. This gradual loss of efficacy is explained by selection in favour of T. acuformis rather than T. yallundae (which had a naturally narrower range of sensitivity levels in its populations) and for strains with least sensitivity within the T. acuformis population, although no resistance to prochloraz was found. Effects of treatments on yield were usually not statistically significant. This may be explained partly by the predominance of the more slowly developing T. acuformis in many of the plots.     

小麦种质抗纹枯病性的鉴定和遗传分析

对2300余份创新种质和引进种质抗纹枯病性进行了自然病地初步鉴定,在此基础上选择一批抗性好的种质进行了人工病圃鉴定,共评选出高抗纹枯病的创新种质14份,引进种质21份。这些种质都兼抗1－3种其它病害,且综合性状较好,其中创新种质最好,为抗纹枯病育种提供了良好的种质材料。还选用7个抗纹枯病性不同的亲本组配成半双列杂交组合。采用Hayman法进行了基因效应分析,结果表明,抗纹枯病性的遗传不符合加性－显性模型。     

Development of Pseudocercosporella herpotrichoides (Fron) Deighton var. herpotrichoides and var. acuformis on Wheat Plants Measured by ELISA

Using the ELISA method, the development of Pseudocercosporella herpotrtchoides var. herpotrichoides and var. acuformis in a susceptible cultivar of winter wheat was compared under controlled and held conditions. In the greenhouse, var. acufornis grew less vigorously, was slower in penetrating the coleoptile and the successive leaf sheaths and in colonizing the stem tissue than var. herpotrichoides. In the field, these differences were confirmed on the last leaf sheaths and the stem. At ripening stage, however, identical ELISA values were measured for both varieties. Moreover, a significant variation was observed between the individual isolates of each variety. Comparison of the effect of both varieties of P. herpotrichoides on 20 wheat cultivars characterized by different resistance levels showed significant interactions. The cultivars carrying the Pch-1 gene always remained the less diseased genotypes. In general, var. acuformis developed less antigen in the cultivars than var. herpotrichoides. It is concluded, that in tests for resistance to P. herpotrichoides mixtures of many highly pathogenic isolates of both fungus varieties should be used. Less complex mixtures or single isolates may result in wrong estimates of resistance.     

The role of antibiotic production by a strain of Pseudomonas fluorescens in the suppression of Pseudocercosporella herpotrichoides, the causal agent of eyespot disease of cereals

Pseudomonas fluorescens strain 220 is an effective antagonist of Pseudocercosporella herpotrichoides , the eyespot pathogen of cereals. Culture filtrates of Ps. fluorescens 220 were inhibitory to spore germination and hyphal growth of P. herpotrichoides and at least two compounds with antifungal and antibacterial activity were identified in cultures grown in nutrient broth. In plant tests, both a culture broth of Ps. fluorescens 220 and a crude antibiotic extract reduced eyespot disease, whereas a mutant strain of 220 deficient in antibiotic production had no effect. Production of antibiotics would therefore appear to be a major factor in the suppression of P. herpotrichoides infection. A loss of disease control when Ps. fluorescens 220 was applied to plants in water was not due to lack of survival, as populations of a marked strain of Ps. fluorescens 220 applied to the stem base of wheat plants were similar whether applied in water or culture broth.     

Studies on Mechanisms of Splash Dispersal of Spores, using Pseudocercosporella herpotrichoides Spores

Simulated raindrops, 4 or 5 mm in diameter, fell 13 m onto target water films, with Pseudocercosporella herpotrichoides spores incorporated into either drops or targets. Resulting splash droplets were collected on fixed photographic film and numbers of droplets, spore-carrying droplets and spores determined.
The patterns of dispersal of splash droplets, spore-carrying droplets and spores with distance and droplet size were similar for 4 mm and 5 mm incident drops with spores incorporated into either targets or drops. Numbers of droplets, spore-carrying droplets and spores decreased with increasing distance from targets and none were collected at 1 m. However, more spores were dispersed by 5 mm than by 4 mm drops and with spores in targets than with spores in incident drops. Whereas most splash droplets were in the smallest size category (0–100 μm), most spore-carrying droplets were 200–400 μm and most spores were in droplets with diameter greater than 1000 μm. Regressions of square root (number of spores) on droplet diameter were significant (p < 0.001) in all cases. The slopes of regression lines were greater when spores were in targets than when they were in incident drops. Splash droplets were collected up to a height of 70 cm, with most between 15 and 20 cm. The dye experiment showed that most splash droplets contained liquid from both incident drop and target film.     

Investigations on the Activity of Cell Wall-degrading Enzymes in Young Wheat Plants After Infection with Pseudocercosporella herpotrichoides (From) Deighton

In earlier investigations, it has been demonstrated that Pseudocercosporella herpotrichoides (Fron) Deighton is capable of producing pectolytic and cellulolytic enzymes as well as hemicellulases in vitro. The investigation of enzyme activity in extracts from wheat plants infected with P. herpotrichoides (isolates 21e and R6) and from non-infected plants revealed the activity of the following enzymes: pectin methylesterase (PME), polymethylgalacturonase (PMG), pectin lyase (PL), carboxymethylcellulase (CMCase), xylanase and arabanase. Compared to non-infected plants, the enzyme activity in infected plants was considerably higher; in some experiments, only traces of enzyme activity could be found in control plants. The difference in the enzyme activity in infected as compared to non-infected plants was, in most cases, statistically significant, especially beginning at the end of the second week after inoculation.
The enzyme activity depended on the temperature during plant cultivation; with the exception of pectin methylesterase (PME), the activity of all investigated enzymes increased with temperature and the highest activity was found in plants grown at 20°C. The highest PME activity was measured in plants grown at 10°C; the activity of this enzyme was generally lower at 15 and 20°C.     

Influence of direct-drilling of winter wheat on incidence of take-all and eyespot

Winter wheat drilled directly into stubble or pasture treated with paraquat to kill the vegetation has been found to be less severely attacked by take-all (Ophiobolus graminis (Sacc.) Sacc.) and eyespot (Cercosporella herpotrichoides Fron) than wheat drilled after cultivation. The reduction of take-all is associated, not with a direct effect of the chemical, but with factors, resulting from the technique, which limit the rate of spread of the fungus in the undisturbed soil.