Experiments on soil drenching with fungicides against take-all in wheat

In short term pot experiments benomyl, iprodione and KWG 0599 applied as soil drenches in several types of soil significantly suppressed take-all symptoms from inoculum placed just below wheat seeds planted 1×5 cm deep, and in sand but not other soils when seeds were 5 cm deep. Benomyl was, however, effective against inoculum below seed planted 5 cm deep in a loam-sand mixture when the drench contained an alcohol ethoxylate surfactant. Computer simulations of fungicide distributions in the soils correlated well with disease control observations. In long term outdoor pot experiments two drenches with benomyl (without surfactant) controlled disease significantly for at least 3 months against inoculum placed 15 cm deep. The significance of these results for the practical control of take-all by fungicides is discussed.     

Effects of wheat volunteers and blackgrass in set-aside following a winter wheat crop on soil infectivity and soil conduciveness to take-all

Two experiments were carried out in France in which disease indices were used to evaluate the effects of wheat volunteers and blackgrass (Alopecurus myosuroides) on soil infectivity and soil conduciveness to take-all caused by Gaeumannomyces graminis var. tritici. Soil infectivity was evaluated by measuring the disease index on susceptible wheat plants grown on soil samples collected from the field. Soil conduciveness to the disease was obtained by measuring disease indices on plants grown on soil samples to which different amounts of take-all fungus inoculum were added. One experiment (Expt. 1) was carried out using soils from farmers' fields (two fields in 1994 and two in 1995); soil infectivity and soil conduciveness were evaluated for three experimental situations: bare soil, soil with wheat volunteers and soil with blackgrass plants. In 1994 the soil infectivity was zero in bare soil, high with the wheat cover, and intermediate with the blackgrass cover. In 1995 the soil infectivity was uniformly low for all three conditions. Soils bearing wheat were less conducive than bare soil, soils bearing blackgrass and bare soils were similarly conducive. A second experiment (Expt. 2) carried out in 1995 compared the soil infectivity and soil conduciveness to take-all of soils planted with wheat or blackgrass in set-aside land after periods of wheat monoculture of 0–6 yr. The soil infectivity was low for all treatments. The soil was more conducive after blackgrass than after wheat. In both cases, the soil conduciveness was less when the monoculture had continued for more than 4 yr. The decline was less after blackgrass than after wheat. Thus, whenever set-aside is set up during the increase phase of the disease in fields with cereal successions, abundant wheat volunteers might hinder the expected positive effect of a break in cereal successions on take-all development. The presence of blackgrass in a set-aside field, with significant soil infectivity and high soil conduciveness, might increase the risks of take-all development in a wheat crop following set-aside.     

Evaluation of fungicides applied to soil to control naturally-occurring take-all using a balanced-incomplete-block design and very small plots

Six sterol biosynthesis-inhibiting fungicides representing several combinations of properties were applied to soil to control naturally-occurring take-all (caused by Gaeumannomyces graminis var. tritici) in winter wheat in field experiments in two successive years. The average take-all severity category was never more than moderate in the different clay-loam and sandy loam sites used in each year. At each site in each year there were six treatments and an untreated control in an arrangement based on a balanced-incomplete-block design for six treatments in 10 blocks each with three treatments. Each block had three treated plots and a control plot and was paired with the complementary block of three treatments (plus control) to form a complete replicate, of which there were 30 per site. Take-all assessments in June or July showed that after incorporation into the seed bed (at 2 kg ha“¹and sometimes at 1 kg ha”¹) in autumn, two non-volatile, strongly lipophilic compounds, nuarimol and triadimenol, with good intrinsic toxicity to the take-all fungus and slow rates of degradation, partially controlled take-all. However, another compound, flutriafol, with similar properties to nuarimol and triadimenol, controlled take-all less. Two slightly volatile, strongly lipophilic compounds, flusilazole and penconazole, with good intrinsic activity, were less effective (at 2 kg ha^-1). A volatile, less lipophilic compound, PP 969, with less intrinsic activity, also partially controlled take-all, but only after application as a drench in the spring (2 kg ha^-1). The most effective treatments were generally more effective the greater the level of disease (as indicated by assessments of disease in control plots), especially in spring assessments of disease. Although flutriafol did not perform as expected, it still seems reasonable to conclude that the requirements for a soil-applied fungicide to control take-all are likely to be: (i) good intrinsic fungitoxicity, (ii) some mobility in soil water (i.e. not strongly lipophilic), and (iii) season-long persistence.     

Natural suppression of take-all disease of wheat in Montana soils

This research was initiated to determine whether soils suppressive to take-all of wheat caused by Gaeumannomyces graminis var. tritici (Ggt) occur in Montana, and to identify the organisms most likely involved in this suppression. From an initial screening of eight soils collected from different wheat growing areas of Montana, two were highly suppressive to take-all. Microbial characterization of these soils indicated that different mechanisms were involved in the suppression. In Larslan soil, mycoparasitism appeared to be the main mechanism. Two different fungi with exceptional ability to reduce the severity of take-all were isolated from this soil. One of these fungi could parasitize the hyphae of Ggt. Field tests with these fungi in Ggt infested soil showed increases of over 100% in both harvestble tillers and grain yield as compared to treatments without these two fungi. In tests with 48 different bacteria and 10 actinomycetes from Larslan soil, none were able to consistently reduce severity of take-all alone, or in mixtures. In Toston soil, antibiosis by actinomycetes and perhaps the involvement of Pseudomonas spp. in production of antibiotics and/or siderophores appeared to be the most likely mechanisms involved in take-all suppression. Increases in shoot dry weight over that in the Ggt infested control using mixtures of pseudomonads and actinomycetes ranged from 25% to 87%. Actinomycetes added individually or in mixtures to soil infested with Ggt consistently reduced the severity of the disease to a greater extent than did mixtures of Pseudomonas spp.     

The microbiology of soils under successive wheat crops in relation to take-all disease

Abstract Thirty-eight wheat fields in southern England were sampled in an attempt to correlate the amount of take-all disease with 35 microbiological and chemical measurements of soil. There was little correlation between field take-all and pot tests to determine soil infectivity. Myxogastrids were important components of the soil population, being up to half of the amoebal population, and most soils contained dictyostelids, reticulate amoebae and myxobacteria. Amoebae, ciliates, bacteria and saprophytic fungi were recorded for all soils. pH was a major determinant of soil populations, being clearly correlated with fungal abundance and with numbers of ciliates, dictyostelids and bacteria. Principal component analysis separated dictyostelids from the other soil amoebae and again showed the importance of pH in determining soil microbial populations. Take-all was negatively correlated with soil fertility and positively related to nematodes and myxobacteria, but this was probably an effect of take-all, and represented saprophytic growth on dead roots rather than being a cause. Reticulate amoebae and dictyostelids were both correlated with low levels of take-all. This study emphasises the large number of interrelated populations of soil microorganisms which could have an effect on the severity of take-all infections.     

Effects of ammonium-N on development and infectivity of the take-all fungus

Effects of applications of a mixture of ammonium sulphate and mono-ammonium phosphate and of ammonium nitrate on the incidence of take-all disease of wheat (caused by Gaeumannomyces graminis var. tritici) and on subsequent inoculum levels were studied in field and glasshouse experiments. In a field experiment in Western Australia, on a sandy soil at pH 5·4, nitrogen applications had no detectable effect on disease severity at anthesis, but ammonium sulphate treatment increased the number of propagules of the pathogen in the soil. In a pot experiment, in which seed was sown in the field experiment soils, disease was greater in soil from plots treated with ammonium sulphate and least in soil from the nil-nitrogen plots, reflecting the respective inoculum levels in the field plot. However, treatment of the soils of lower inoculum with ammonium sulphate and ammonium nitrate during this pot experiment decreased disease. A second pot experiment confirmed the effectiveness of ammonium sulphate and ammonium nitrate in reducing take-all at lower inoculum levels, and their ineffectiveness at higher inoculum levels.     

Plant genotype, micronutrient fertilization and take-all infection influence bacterial populations in the rhizosphere of wheat

The relationship between micronutrient efficiency of four wheat (Triticum aestivum L.) genotypes, tolerance to take-all disease (caused by Gaeumannomyces graminis (Sacc.) Arx and Olivier var. tritici Walker), and bacterial populations in the rhizosphere was tested in soil fertilized differentially with Zn and Mn. Plant growth was reduced by Mn or Zn deficiency and also by take-all. There was an inverse relationship between micronutrient efficiency of wheat genotypes when grown in deficient soils and the length of take-all lesions on roots (efficient genotypes had shorter lesions than inefficient ones). In comparison to the rhizosphere of control plants of genotypes Aroona and C8MM receiving sufficient Mn and Zn, the total numbers of bacterial cfu (colony forming units) were greater in the rhizosphere of Zn-efficient genotype Aroona under Zn deficiency and in Mn-efficient genotype C8MM under Mn deficiency. These effects were not observed in other genotypes. Take-all decreased the number of bacterial cfu in the rhizosphere of fully-fertilized plants but not of those subjected to either Mn or Zn deficiency. In contrast, the Zn deficiency treatment acted synergistically with take-all to increase the number of fluorescent pseudomonads in the rhizosphere. Although numbers of Mn-oxidising and Mn-reducing bacteria were generally low, take-all disease increased the number of Mn reducers in the rhizosphere of Mn-efficient genotypes Aroona and C8MM. Under Mn-deficiency conditions, the number of Mn reducers in the rhizosphere increased in Aroona but not in C8MM wheat. The results suggest that bacterial microflora may play a role in the expression of Mn and Zn efficiency and tolerance to take-all in some wheat genotypes.     

Relationships between take-all,soil conduciveness to the disease,populations of fluorescent pseudomonads and nitrogen fertilizers

Take-all of wheat, caused by Gaeumannomyces graminis var tritici (Ggt), is reduced by ammoniacal fertilizers as compared to nitrate sources. This influence of nitrogen on the disease is only observed on nodal roots at flowering. But soil conduciveness to take-all, as measured in a soil bioassay, is modified earlier. Forty days after nitrogen application at early tillering, the NH₄-treated soil became less conducive than the NO₃-treated one. When nitrogen applications are done at sowing and at tillering, differences in disease propagation between the two soils are enhanced. Results from four years of experimentation show that when the level of natural soil inoculum is high, disease severity is reduced by ammonium, showing an effect on the parasitic phase of Ggt. At a low level of natural inoculum the effect of the source of nitrogen is mainly observed on the percent of infected plants, indicating that the saprophytic and preparasitic phases are affected. Rhizospheric bacterial populations increase from sowing to tillering, but differences on take-all conduciveness after tillering are not correlated with differences in the amounts of aerobic bacteria or fluorescent pseudomonads isolated from soils treated with different sources of nitrogen. Qualitative changes in fluorescent Pseudomonas spp. populations, like in vitro antagonism, are more likely to explain differences in soil conduciveness to take-all than are quantitative changes in this group. Nevertheless, the introduction of Ggt in a cropped soil leads to a greater increase in fluorescent pseudomonads populations than in total aerobic bacteria.The delay between reducing soil conduciveness and reducing disease in the field with ammonium nitrogen fertilization, the qualitative change of fluorescent pseudomonads populations and the role of necroses in rhizobacteria multiplication, provide information leading to our representation of a dynamic model based on the differentiation of the wheat root system into seminal and nodal roots.     

Soil-applied fungicides for controlling take-all in field experiments with winter wheat

Soil treatment fungicides were tested against take-all (Gaeumannomyces graminis var. tritici) in three field experiments with winter wheat. Fungicides were applied as drenches either before sowing in autumn, and incorporated by rotary harrowing, or to the crop in spring. The most effective treatments were autumn applied benomyl (20 kg/ha) and nuarimol (0·55-4·4 kg/ha). However, the highest nuarimol concentration depressed yield. Benomyl sometimes induced a resurgence of take-all in the second wheat crop after treatment. Nuarimol had no adverse effects in subsequent crops, and neither fungicide hindered the onset of take-all decline in a third crop after treatment. The possible value of soil treatment in future control strategies is discussed.     

Intermittent wetting of soils at high temperature reduces survival of the take-all fungus

Two pot experiments using naturally infested soil and a range of watering regimes were conducted to study the possible effect of level and frequency of wetting of hot soil (to simulate the period between growing seasons in Western Australia) on inoculum of the take-all fungus (Gaeumannomyces graminis var.tritici). In combination with the high soil temperatures, all watering regimes reduced infectivity and propagule number of the take-all fungus, this reduction being absent in dry soils.     

Soil conduciveness to take-all of wheat: Influence of the nitrogen fertilizers on the structure of populations of fluorescent pseudomonads

In a field cropped with wheat, a high and low level of soil conduciveness to take-all were induced by applying a nitrogen fertilizer with either calcium nitrate or ammonium sulphate. From these two soils, two representative populations of fluorescent pseudomonads were tested for their in situ behaviour. Take-all index and root dry weight were assessed on plants cropped in soils infested with Gaeumannomyces graminis var tritici (Ggt) and each bacterized with one of the isolates of fluorescent pseudomonads. The bacteria tested can be split into three groups: antagonists which reduce take-all, deleterious isolates which aggravate the disease and neutral without evident effect on the disease. The predominance of antagonistic fluorescent pseudomonads in the NH₄-treated soil and the predominance of deleterious ones in the NO₃-treated soil was confirmed after statistical analysis. The microbial impact on take-all must be more considered as the resulting effect of divergent activities of both rhizobacteria types than the only consequences of the presence of antagonistic pseudomonads. All the high cyanogenic pseudomonads were antagonists in situ and were more numerous in the NH₄-treated soil than in the NO₃-treated soil.     

Prediction of take-all disease risk in field soils using a rapid and quantitative DNA soil assay

A rapid, routine DNA-based assay to quantify Gaeumannomyces graminis var. tritici (Ggt), the causal agent of take-all disease of cereals, has been developed and used for the prediction of take-all in a wide range of field soils. Based on the correlation of the DNA-based assay and a soil bioassay, the risk of disease development can be estimated. Ggt DNA levels of <30 pg, 30–50 pg and >50 pg in 0.1 g soil organic matter correspond to low, moderate and high levels of the disease, respectively. Limitations in the prediction of take-all, including sampling requirements to obtain representative soil samples from fields and increasing the sensitivity and the accuracy of the DNA assay, are described. The main advantage in using the DNA-based assay, in estimating the amount of Ggt inoculum in soil, is that the levels of Ggt in soil samples can be assessed rapidly and accurately. Farmers can now have soil samples assessed before sowing. The DNA result can be used to predict the potential yield loss and determine the most appropriate management options using decision support software that is currently available. This DNA technology is currently being used commercially to detect and predict take-all. This revised version was published online in June 2006 with corrections to the Cover Date.     

An experimental design and procedures for testing putative controls against naturally-occurring take-all in the field

In experiments during 1983–86 take-all was more severe and eyespot and sharp eyespot less frequent in 2nd-4th crops of winter wheat at Woburn (Beds.) than at Rothamsted (Herts.). Third crops had most take-all and yielded least grain. Against this background, small plots, 37 cm × 31 cm, in which all plants were sampled, were tried as a means of increasing experimental precision. They were arranged in fours in incomplete blocks and blocks with complementary treatments (putative controls of take-all) were paired. Thirty of these block-pairs were distributed throughout each experimental site in each year to provide one replicate of the design for each of three sampling times: April, June and August. Unattributed variation in disease and plant growth for plots within blocks was compared to that in other strata (block-pairs and blocks within block-pairs) of the experiment. The variability amongst block-pairs scattered throughout the site was nearly always greater than that for blocks within block-pairs (98% of take-all assessments, 71% of soil infectivity estimates, 94% of eyespot and sharp eyespot assessments and 86% of all plant measurements). The variability of blocks within block-pairs exceeded that of plots much less frequently (56% and 69% of take-all assessments, 33% and 25% of soil infectivity measurements, 63% and 56% of eyespot and sharp eyespot assessments and 50% and 63% of plant measurements; Rothamsted and Woburn, respectively). Small plots were judged mostly on this last comparison, where a variance ratio in excess of 1 indicated that the small plots had decreased variability and increased precision. Variance ratios for different assessments of take-all indicates that small plots: i) most consistently decreased disease variability during the years of maximum disease, ii) were slightly less effective at Rothamsted than at Woburn, and iii) were usually less effective in fourth crops than in previous crops. Soil infectivity was most uniform after crops with most disease and blocks were rarely more variable than plots. hxcept when disease was severe, soil infectivity in August tended to be positively associated with the yield of the crop just harvested. These findings reveal changes in the scale of disease patterns, both during the crop sequence and within individual crops, and suggest more than one scale of pattern in take-all-infested fields. This is discussed in relation to field experimentation and take-all decline.     

Prospects for fungicidal control of take-all of wheat

A range of fungicides and herbicides was tested against Gaeumannomyces graminis (causal agent of take-all) on agar plates, and on wheat seedlings in pots and in liquid culture. Benomyl, the standard in all tests, was consistently most effective: like iprodione, nuarimol and KWG 0599 , it diminished infection from inoculum placed just below the seeds more effectively when applied as a drench than as a soil-mix. Benomyl as a soil-mix was most effective in soils with least organic matter. Some compounds toxic to the pathogen on agar plates and in plants grown in liquid culture were ineffective as soil treatments. The practical limitations of soil treatment with conventional fungicides and application methods are discussed.     

Comparison of natural and artificial epidemics of take-all in sequences of winter wheat crops

Winter wheat was grown for six successive years (Expt 1) and for three successive years (Expt 2) in field experiments on different soil types. Artificial inoculum of the take-all fungus (Gaeumannomyces graminis var. tritici cultured on autoclaved oat grains) was incorporated in the soil of some of the plots just before, or at, sowing of the first winter wheat crop. Expt 1 tested the incorporation of similar amounts of inoculum (212 kg ha^-1) at different depths. Expt 2 tested different amounts of inoculum at the same, shallow depth. Early sowing (September), late sowing (October) and spring inoculation were additional treatments, applied to the first crop only, in Expt 2. Seasonal factors apart, the disease outcome in the first year after inoculation depended on amounts and placement of applied inoculum, as well as date of sowing. Deeper inoculum resulted in less disease (Expt 1). Severe take-all was produced in Expt 2 by incorporating inoculum shallowly in sufficient quantities (400 kg ha^-1 or more). Less inoculum (200 kg ha^-1) generated less disease, especially in earlier-sown plots. Differences in disease amongst inoculum treatments were greatest in the first year and diminished subsequently, particularly where sowing had been early in the first year. In Expt 1, where first crops exposed to artificial inoculum developed moderate-to-severe disease, disease in subsequent second and/or third crops was less. In the fourth crop a second peak of disease occurred, coinciding with a first peak in sequences without added inoculum. Take-all decline (TAD) appeared to be expressed in all sequences thereafter. In Expt 2 in sequences without added inoculum, TAD occurred after a peak of disease in the second crop. Where 400 kg ha^-1 or more of inoculum were added, disease was severe in the first year and decreased progressively in successive years. Disease was less patchy in plots that received artificial inoculum. However, it remains uncertain mat severe disease caused by artificial inoculation achieved an early onset of true TAD. The infectivity of the top 12 cm of soil in the first 3 yr of Expt 1, determined by bioassay, depended on the depth of added inoculum and amount of disease in subsequent crops. However, at the time of the naturally occurring peak of disease severity (in either inoculated or non-inoculated plots) it did not predict either disease or TAD. Differences and similarities amongst epidemics developing naturally and those developing from different amounts and placement of applied inoculum have been revealed. The epidemiological implications of adding inoculum and the potential value of artificially-created epidemics of take-all in field trials are discussed.     

Use of a root assessment tray for the detection of take-all on lateral roots of wheat seedlings

A root assessment tray was designed for the meticulous assessment of take-all on wheat seedling roots from soil bioassays. Subsequently, the detection of lateral root infections (in addition to the more obvious infections on main axes of seminal roots) resulted in increased estimates of propagule numbers of the take-all fungus (Gaeumannomyces graminis var.tritici) for 196 of the 368 soil samples bioassayed in a field study conducted in Western Australia between 1984 and 1986.     

Field tests of bacteria and soil-applied fungicides as control agents for take-all in winter wheat

Putative biological and chemical treatments for controlling take-all were used in each of three consecutive years at two locations where winter wheat crops were grown in naturally-infested fields. The chemical treatments more often decreased take-all than the biological treatments, but no treatment consistently and significantly decreased take-all, nor did any cause a significant increase in yield. An isolate of Bacillus cereus var. mycoides and one of B. pumilis, applied as soil drenches in autumn or spring, or in the seed furrows, were usually ineffective. Of the few significant effects on disease, half were associated with increases and half with decreases, and most occurred in April and did not persist to late June. Two strains of Pseudomonas pluorescens applied to the seed were ineffective. The fungicide benomyl, applied as a drench in autumn and spring at 20 kg/ha was ineffective, while nuarimol, applied as a drench in autumn at 2 kg/ha was sometimes effective. Nuarimol incorporated into the seed bed at 2 kg/ha was the most effective treatment. In analyses using a functional relationship model for data from treated and untreated plots 12% of 176 data sets for biological treatments, 38% of 96 data sets for chemical treatments and 81% of 16 data sets for combined treatments showed increasing efficiency of the treatment with increasing disease intensity. These findings also demonstrate an additional advantage of the experimental design, namely that treatments are tested at different disease intensity levels within fields.     

Methods of investigating populations of the take-all fungus (Ophiobolus graminis) in soil

Attempts to isolate Ophiobolus graminis directly from infested soils failed, so host-infection techniques were used to study soil-borne populations of the fungus. Extracting organic debris from soils and grading it by wet sieving through standard meshes concentrated the fungus. Fractions were tested for infectivity either as layers in pots of sand or by packing into short lengths of polyvinyl chloride tubing, through which wheat seedlings were grown. Coarse debris (retained by 420 μ aperture sieves) was most infectious and usually caused lesions within 3 weeks; whole soil and especially fine debris (< 420 μ) caused fewer and less severe infections, which often became apparent only after 3 weeks. Slight infectivity of the sediment fractions was attributed to imperfect separation of debris. Soil sampled in crop or stubble rows caused more infections than soil from between rows. Usually seedling infection was made manifest by root lesions and runner hyphae, but these symptoms were not always plain or conclusive. Many seedling roots that rotted when kept moist and in the light produced perithecia within 6 weeks. Although perithecia formed on some roots where neither lesions nor hyphae were found, they did not form on all diseased roots. More needs to be known about the factors influencing perithecial formation before it can be used as a reliable confirmatory test.     

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.     

Gluconic acid: an antifungal agent produced by Pseudomonas species in biological control of take-all

Pseudomonas strain AN5 (Ps. str. AN5), a non-fluorescent Australian bacterial isolate, is an effective biological control (biocontrol) agent of the take-all disease of wheat caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Ps. str. AN5 controls Ggt by producing an antifungal compound which was purified by thin layer and column chromatography, and identified by NMR and mass spectroscopic analysis to be d-gluconic acid. Commercially bought pure gluconic acid strongly inhibited Ggt. Two different transposon mutants of Ps. str. AN5 which had lost take-all biocontrol did not produce d-gluconic acid. Gluconic acid production was restored, along with take-all biocontrol, when one of these transposon mutants was complemented with the corresponding open reading frame from wild-type genomic DNA. Gluconic acid was detected in the rhizosphere of wheat roots treated with the wild-type Ps. str. AN5, but not in untreated wheat or wheat treated with a transposon mutant strain which had lost biocontrol. The antifungal compounds phenazine-1-carboxylic acid and 2,4-diacetylphloroglucinol, produced by other Pseudomonads and previously shown to be effective in suppressing the take-all disease, were not detected in Ps. str. AN5 extracts. These results suggest that d-gluconic acid is the most significant antifungal agent produced by Ps. str. AN5 in biocontrol of take-all on wheat roots.