Uptake and distribution of [14C]metalaxyl by detached tobacco leaves

When the petioles of detached tobacco leaves (10–17 cm²) were incubated in aqueous solutions containing [¹⁴C]metalaxyl, uptake of the fungicide was dependent on the temperature and photoperiod. Detached leaves took up 78% more [¹⁴C]metalaxyl at 26°C than at 16°C. The rate of uptake in the light at 21°C was linear, but after an additional 20h in the dark, there was only twice as much fungicide in the leaves. Different sized leaves contained the same amount of fungicide per cm² area. Uptake by detached leaves of the ¹⁴C-labelled anilide lactones ofurace and RE-26940 [2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)acet-2′,6′-xylidide] was similar to that of metalaxyl. At the concentration of metalaxyl (66 ng ml^?1) that controlled blue mould (Peronospora tabacina) on detached tobacco leaves, the amount of fungicide in the leaves was found to be 7.25 ng. Autoradiography showed that the distribution of [14C]metalaxyl in detached leaves after incubation for 23h was uniform, although higher concentrations of the label were present in the smaller veins of the leaves.     

Resistance to acylalanines in Pseudoperonospora cubensis1

Resistance to acylalanine fungicides in Pseudoperonospora cubensis, the downy mildew of cucurbits, is reviewed. Insensitive strains of the fungus predominated during the second year of metalaxyl application in cucumber glasshouses in Greece and Israel. Resistance was detected either on detached cucumber leaves floated on metalaxyl solutions or on potted plants treated (sprayed or drenched) with this fungicide. Metalaxyl-resistant strains of P. cubensis showed good pathogenicity and fitness, and competed favorably with the sensitive strains in the absence of the fungicide. A synergism between these two biotypes resulted in an increased virulence of the resistant forms on metalaxyl-treated plants. Metalaxyl-resistant strains of the pathogen exhibited cross resistance to other acylalanine fungicides. Strains resistant to acylalanines and to the chemically unrelated oomycete-fungicides fosetyl-Al and propamocarb were isolated in Israel, as well. In Greece, however, protective sprays with fosetyl-Al, cymoxanil and mancozeb gave good control of cucumber downy mildew when the pathogen was resistant to metalaxyl. P. cubensis is the first case of fungus which developed resistance to acylalanines in the field.     

多菌灵等4种杀菌剂对烟草灰霉病菌的室内生物活性

采用菌丝生长速率法和孢子萌发法,分别测定了烟草灰霉病菌对多菌灵、嘧霉胺、异菌脲和丙环唑的敏感性,同时通过离体叶片法评估了这4种杀菌剂对烟草灰霉病的保护和治疗作用。结果表明:4种杀菌剂对烟草灰霉病菌的菌丝生长和孢子萌发均表现出了不同程度的抑制活性,并对灰霉病同时具有保护和治疗作用。其中多菌灵对菌丝生长的抑制活性最强,EC₅₀平均值为0.06 mg/L,其次为丙环唑、嘧霉胺和异菌脲,EC₅₀平均值分别为0.36、0.53和0.60 mg/L;异菌脲和丙环唑对烟草灰霉病菌孢子萌发的抑制活性较强,EC₅₀平均值分别为2.05和2.21 mg/L,其次为嘧霉胺和多菌灵,EC₅₀平均值分别为10.56和131.23 mg/L。异菌脲和多菌灵对灰霉病的保护作用和治疗作用均最强,当药剂质量浓度为200 mg/L时,其对离体叶片的保护和治疗作用防效分别为100%、100%和98.3%、91.8%。研究结果可为烟草灰霉病的科学防治提供依据。     

Studies on metalaxyl-resistant Phytophthora infestans in potato crops in south-west Scotland

In 1981 metalaxyl-resistant Phytophthora infestans was found in 44% of the 63 crops examined in south-west Scotland. In 1982, when fungicides containing acylalanines (metalaxyl or ofurace) were not used in the area, it was detected in 15%. Metalaxyl-resistant strains were found in several crops which had been sprayed only with non-acylalanine fungicides or had received no fungicidal treatment. In neither year was resistance associated with a significant breakdown of disease control.
Metalaxyl-resistant isolates of the blight fungus were obtained from tubers from crops in which resistance was detected on the foliage.
In detached leaf experiments, resistant isolates retained their ability to grow on leaves treated with 100 μg/ml of metalaxyl after 52 weekly transfers on untreated leaves, and produced sporangia on leaves treated with up to 2000 μg/ml of metalaxyl or ofurace. In mixtures with sensitive sporangia the proportions remained stable after nine weekly transfers on untreated leaves. However, at concentrations as low as 50 μg/ml, metalaxyl and ofurace reduced the extent of colonization and the numbers of sporangia produced by resistant isolates by an average of 50%.     

SIMPEROTA 1/3 – a decision support system for blue mould disease of tobacco*

Blue mould (Peronospora tabacina) is the most serious threat to German tobacco crops. In order to efficiently control the disease whilst minimizing the risk of nontolerable fungicide residue levels on tobacco leaves, a decision support system has been developed which optimizes the timing of fungicide treatments. The DSS consists of two models, SIMPEROTA 1, which forecasts the dates of blue mould first appearance and SIMPEROTA 3 which forecasts the dates of fungicide applications. Crucial biological processes are included in the models (infection, mycelium growth, sporulation and spore release). Input parameters are temperature, relative humidity and leaf wetness recorded on an hourly basis. Validation with data from 2003 and 2006 showed that SIMPEROTA 1 gave satisfying results. The model is suitable for practical use and can be employed for steering monitoring efforts of extension services and for the timing of the first fungicide treatment. SIMPEROTA 3 gives advice on follow‐up treatments and the length of spraying intervals, but this model needs to be validated before being introduced into practice.     

Integrated pest management: A strategy to control resistance of Spodoptera exigua and Helicoverpa armigera caterpillars to insecticides on soybean in the Mekong Delta

The antibiotic nucleoside tubercidin produced by Streptomyces viola-ceoniger was evaluated for in-vivo efficacy and in-vitro activity against Phytophthora capsici, Magnaporthe grisea and Colletotrichum gloeosporioides. Tubercidin was more effective against P. capsici and M. grisea than against C. gloeosporioides in inhibiting mycelial growth. The bioassay on TLC plates was the most sensitive method and allowed the evaluation of antifungal activity of tubercidin even at a low concentration of 0.1 μgml^?1. As compared to the systemic fungicide metalaxyl, tubercidin was similar or somewhat higher in inhibition of mycelial growth of P. capsici. When applied to pepper stems, tubercidin was equally as effective as metalaxyl in the control of phytophthora blight in pepper plants, irrespective of application time and concentration. The treatment with 1000 μg ml^?1 tubercidin induced phytotoxicity in pepper plants. No control efficacy of phytophtora blight was observed in pepper plants supplied with a soil drench of tubercidin. Treatment with tubercidin at 500 μg ml^?1 completely protected pepper plants at first branch stage from phytophthora blight until four days after application. The control efficacy of tubercidin drastically declined seven days after application.     

PCR-based monitoring of recent isolates of tobacco blue mold from Europe reveals the presence of two genetically distinct phenotypes differing in fungicide sensitivity

Bioassays testing the fungicide sensitivity against metalaxyl of Peronospora tabacina isolates collected in German tobacco fields in 2005 revealed the presence of two phenotypes, resistant and sensitive. DNA fingerprints using SSR and minisatellite primers allowed separation of the samples into two groups. The differences in amplification patterns coincided with the sensitive and resistant reaction of the isolates in metalaxyl bioassays. New primers were developed which allowed PCR-based detection of P. tabacina and differentiation of the metalaxyl-sensitive and the metalaxyl-resistant phenotype, respectively. Screening of recent blue mold isolates from Germany and other European countries for metalaxyl sensitivity with leaf disk bioassays coincided completely with the PCR-based identification of the two phenotypes. In Germany, exclusively resistant isolates were found between 2002 and 2004. These still dominate. Since 2005 the co-occurrence of sensitive isolates has been shown. No similar monitoring has been done in any other European country. However, we found the resistant phenotype reaction in a French isolate of 2004 and in two out of three Italian isolates in 2007. Two isolates from Poland and Bulgaria in 2007 were sensitive to metalaxyl. For all 58 isolates tested since 2002 the metalaxyl bioassay-based resistance type and the PCR-based tests for sensitive and resistant genotype coincided. Using genotype-specific primers for future population studies may help to trace the sources of the pathogen from which yearly propagation starts.     

Inhibition of Botrytis cinerea growth and suppression of botrytis bunch rot in grapes using chitosan

Chitosan inhibited growth of Botrytis cinerea in liquid culture and suppressed grey mould on detached grapevine leaves and bunch rot in commercial winegrapes. Germination of B. cinerea was completely inhibited in malt extract broth containing chitosan at concentrations greater than 0·125 g L^?1. However, treated conidia were able to infect detached Chardonnay leaves and pathogenicity was not affected, even after incubation for 24 h in chitosan at 10 g L^?1. When added after conidial germination, chitosan inhibited B. cinerea growth and induced morphological changes suggestive of possible curative activity. The effective concentration of chitosan that reduced mycelial growth by 50% (EC₅₀) was 0·06 g L^?1. As a foliar treatment, chitosan protected detached Chardonnay leaves against B. cinerea and reduced lesion diameter by up to 85% compared with untreated controls. Peroxidase and phenylalanine ammonia‐lyase activities were also induced in treated leaves. In vineyard studies, Chardonnay winegrapes exhibited 7·4% botrytis bunch rot severity at harvest in 2007 after treatment with an integrated programme that included chitosan sprays from bunch closure until 2 weeks preharvest, compared with 15·5% in untreated controls and 5·9% with fungicide treatment. In the following season, botrytis bunch rot severity was 44% in untreated Chardonnay at harvest and the integrated programme (21%) was less effective than fungicides (13·8%). However, in Sauvignon blanc winegrapes, the integrated and the fungicide programme each reduced botrytis bunch rot severity to 4% and were significantly different from the untreated control (11·5%). This study provides evidence that suppression of botrytis in winegrapes by chitosan involves direct and indirect modes of action.     

β-1,3-Glucanase, chitinase, and peroxidase activities in tobacco tissues resistant and susceptible to blue mould as related to flowering, age and sucker development

Tobacco becomes naturally resistant to blue mould caused by Peronospora tabacina as it ages. This age-related resistance is correlated to the time of normal floral development and the senescence of lower leaves: however, it is dependent on neither. β-1,3-Glucanase, chitinase and peroxidase activities increase in tobacco with age. These increases correlate with the development of age-related resistance to blue mould and were independent of floral development. Additionally. β-1,3-glucanase, chitinase, and peroxidase activities were higher in leaf tissue from the main stalk resistant to blue mould) as compared to leaf tissue from suckering stems (susceptible to blue mould) on the same plant. Isozyme patterns of β-1,3-glucanase and chitinase in all resistant tissues are typical of those of tissues systemically protected by either stem injection with Peronospora tabacina or foliar inoculation with TMV.     

Application of fungicides to foliage through overhead sprinkler irrigation – a review

Articles on chemigation with fungicides targeting foliage have been reviewed. They included 23 fungicides tested on 10 crops. Many studies compared chemigation to a check treatment, while others also included conventional methods. Chlorothalonil, followed by mancozeb, fentin hydroxide and captafol were the most studied fungicides, while peanut (Arachis hypogaea), potato (Solanum tuberosum), tomato (Lycopersicon esculentum ), and dry beans (Phaseolus vulgaris) were the most studied crops. Center pivot, followed by solid set, were the irrigation systems most frequently used. The minimum volume of water applied by some center pivots (25 000 litre ha⁻¹ ) is 25 times the maximum volume of water used by conventional ground sprayers. The reduction of fungicide residue on foliage caused by the very large volume of water used by chemigation might be offset by the following factors: (1) fungicide application at the time of maximum leaf wetness when fungi are most active, (2) complete coverage of plants, (3) reducing greatly the inoculum on plant and soil surface, (4) better control of some soil pathogens, and (5) more uniform distribution of fungicides by center pivot. Furthermore, chemigation avoids mechanical damage and soil compaction. Additionally, some systemic fungicides seem to be absorbed rapidly by the leaves, by root uptake from the soil, or by both. In general, all fungicides applied through irrigation water can lessen disease severity. However, when compared to conventional methods, chemigation with fungicides can be less, equally or more effective depending on crop, pathogen, disease severity, fungicide and volume of water. For Cercosporidium personatum control on peanuts, application of protectant fungicides through irrigation water is less effective than conventional methods, but the results with some systemic fungicides mixed with non-emulsified oil and applied through a relatively low volume of water (2.5 mm) are encouraging. Important diseases of potato and tomato can be controlled nearly as well by chemigation as by conventional methods without impairing yield. The main advantage of chemigation for these crops is avoiding a large number of tractor trips through the field and reduced costs of fungicide application. Chemigation has also been shown to be a good option for control of white mold [ Sclerotinia sclerotiorum] on dry beans. © 1999 Society of Chemical Industry     

Failure to detect highly resistant strains in dicarboximide resistant field populations of Botrytis cinerea1

Field isolates of Botrytis cinerea with moderate levels of resistance to dicarboximide fungicides (ED50 1.0–4.9 μg ml^?1) and to dicloran were obtained from glasshouses where vinclozolin and iprodione failed to control grey mould. From sensitive and moderatcly-resistant cultures, laboratory isolates were selected on dicarboximide-amended medium, which were highly resistant to these fungicides (ED50 125->3000 μg ml^?1). Conidia of all the resistant isolates germinated well on media amended with 100 μg ml^?1 of the dicarboximides vinclozolin, iprodione, procymidone and myclozolin and with 5 μg ml^?1 of metomeclan. However, the spores of the moderately resistant isolates did not germinate on 100 μg ml^?1 metomeclan while the spores of the highly resistant isolates germinated well. Using media with 100 μg ml^?1 of metomeclan to distinguish between the two phenotypes, no highly resistant strain was detected among 312 resistant samples from five cucumber glasshouses with a high frequency of moderately resistant strains. From air-borne inoculum of five glasshouses with 100% resistant populations, 1604 colonies were recovered on vinclozolin-amended (100 μg ml^?1) medium and none on metomeclan-amended (100 μg ml^?1) medium. It is concluded that strains of B. cinerea highly resistant to dicarboximides are absent from field populations.     

Removal of duvatrienediols from the surface of tobacco leaves increases their susceptibility to blue mold

Dipping leaf strips of greenhouse or field-grown tobacco (Nicotiana tabacum L., cv. Ky-14) plants into acetone for 1 s, prior to inoculation with sporangia of Peronospora tabacina Adam, increased their susceptibility to blue mold. Disease severity and sporangial production on leaf discs from acetone-treated leaves were markedly increased compared to those on discs from untreated leaves. Treatment with acetone also decreased variation in susceptibility of leaves from plants of various ages. Disease severity on discs obtained from attached leaves which were dipped in acetone for 1 s was three times greater up to 15 days after dipping than on discs from leaves that were not dipped. TLC and GLC analyses of the acetone extracts indicated that 95% or more of the major cuticular diterpenoids, α- and β-4,8,13-duvatriene-1,3-diols (DVT), were removed from the surface by dipping for 1 s. These compounds had not reappeared on the leaf surface 15 days after leaves were dipped in acetone for 1 s. Aqueous suspensions of the acetone-soluble constituents as well as authentic DVT, inhibited sporangial germination of P. tabacina (ED₁₀₀ = 25 ppm) and the antifungal activity was accounted for by DVT. When DVT was removed from a leaf surface and added back to the same leaf strip, the resistance of the leaf tissue was restored. As tobacco plants aged, their susceptibility to blue mold decreased and the quantity of DVT on the leaf surface increased. The data support a role for DVT in the resistance of tobacco against blue mold.     

Efficacy of metalaxyl for blue mould control and its persistence on tobacco

Foliar sprays of metalaxyl, benalaxyl, and cymoxanil plus mancozeb gave better control of blue mould (Peronospora tabacina) than mancozeb alone applied as a spray, or metalaxyl applied to the soil. Before flue-curing, the mean values of metalaxyl residues in tobacco leaves were significantly higher from foliar spray treatments (5.09 μg g⁻¹), than from soil treatments (0.93 μg g⁻¹). Residues had decreased after flue-curing (foliar spray treatment 2.51 μg g⁻¹; soil treatment 0.69 μg g⁻¹), especially on samples taken in September. Curing considerably reduced metalaxyl residues in all cases. Residues of mancozeb ranged from 59.5 to 224.2 μg g⁻¹ before flue-curing and from 11.0 to 22.1 μg g⁻¹ after flue-curing. All these residues were calculated on a dry weight basis. Imidazolidine-2-thione (ethylenethiourea) residues from mancozeb were always below the sensitivity limit of the method used.     

Variations among field isolates of Botrytis cinerea in their sensitivity to antifungal compounds

Seventeen field isolates of Botrytis cinerea were compared by determining their radial growth on synthetic media containing various amounts of 21 antifungal compounds. Twelve of these compounds were fungicides that are recommended for the control of Botrytis infections. There were marked differences between the isolates in their sensitivity to the compounds. Individual isolates displayed high levels of resistance to some of the fungicides, including benomyl, carbendazim, iprodione, thiabendazole, thiophanate-methyl, vinclozolin and zineb. The most potent growth inhibitors were benomyl and carbendazim (ED₉₅ values for most isolates <0.1 μg fungicide ml^?1 media), dichlofluanid, iprodione, nystatin, thiabendazole, thiophanatemethyl and vinclozolin (ED₉₅ values for most isolates < 1.0 μg ml^?1), and captan, chlorothalonil, dicloran and thiram (ED₉₅ values for most isolates < 6.0 μg ml^?1). Zineb was much less potent than the other recommended anti-Botrytis fungicides; it was no more effective than carboxin, dinocap and mancozeb (ED₉₅ values for most isolates > 25 μg ml^?1).     

The advantage of tbz + iprodione treatment for control of gray mold decay of celery caused by the heterogenic spore population ofBotrytis cinerea in Israel

A combined TBZ — iprodione treatment was more effective in inhibiting growthin vitro ofBotrytis cinerea isolates obtained from decayed celery than either of the fungicides applied separately. This was exhibited for both TBZ-resistant and TBZ-sensitive isolates. TBZ at 500 (μg ml^-1 plus iprodione at 1000 μg ml^-1 reduced celery decay beyond the reduction obtained by each fungicide alone. When applied prior to inoculation, the combined treatment prevented decay by the TBZ-sensitive/iprodione-resistant isolates and reduced initial decay by the TBZ-resistant/iprodione-sensitive isolates to 3–10% of the level without treatment. Under natural infection conditions iprodione showed better decay control than TBZ, and at 1500 μg ml^-1 it reduced initial decay during prolonged storage to 3% of the no-treatment level. Although TBZ (500 μg ml^-1) or iprodione (1000 μg ml^-1) applied separately reduced decay significantly, the combination of lower concentrations of each fungicide was sufficient to eliminate decay development almost totally. The combined treatment also inhibited decay bySclerotinia sclerotiorum, which contributed 3% of the total soft rot in stored celery.     

Nested PCR‐RFLP is a high‐speed method to detect fungicide‐resistant Botrytis cinerea at an early growth stage of grapes

BACKGROUND: Grey mould caused by the fungus Botrytis cinerea Pers. ex Fr. is one of the major diseases in grapes. The use of fungicides is a simple strategy to protect grapes against B. cinerea disease. However, phenotypes exhibiting resistance to fungicides have been detected in B. cinerea populations. The variation of fungicide‐resistant B. cinerea isolates renders B. cinerea disease control difficult in grapevine fields. RESULTS: The authors have developed a nested polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) method to detect fungicide‐resistant B. cinerea isolates at an early growth stage of grapes in grapevine fields. The nested PCR‐RFLP method was carried out to detect benzimidazole‐, phenylcarbamate‐ and/or dicarboximide‐resistant B. cinerea isolates from grape berries and leaves at Eichorn–Lorenz growth stage 25 to 29. This method successfully detected fungicide‐resistant B. cinerea isolates at an early growth stage of grapes. In addition, only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates. CONCLUSION: It is proposed that the early diagnosis of fungicide‐resistant B. cinerea isolates would contribute to further improvement of integrated pest management against B. cinerea in grapevine fields, and that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for this purpose. Copyright © 2008 Society of Chemical Industry     

Studies on the systemicity of [14C]metalaxyl in cowpea (vigna unguiculata (L.) walp.)

[¹⁴C]metalaxyl applied to seeds, roots and leaves of cowpea was readily taken up by all plant parts and a major fraction of it was retained within the treated leaves and seeds but not in the roots. The fungicide exhibited ambimobility inside the plant. When applied to the middle of leaflets, metalaxyl mobilised towards proximal and distal parts of the leaflet. Intraleaflet movement seemed to result from translocation through veins and diffusion through cell walls and intercellular spaces. Accumulation of metalaxyl at tips and margins of primary leaves followed application to roots. Metalaxyl was readily extracted from most plant parts except cotyledons and only 0.8% of the fungicide applied as a seed treatment was recovered from 50-day-old plants.     

Detachment of sprayed colloidal copper oxychloride–metalaxyl fungicides by a shallow water flow

BACKGROUND: Flow shear stress induced by rainfall promotes the loss of the pesticides sprayed on crops. Some of the factors influencing the losses of colloidal‐size particulate fungicides are quantified by using a rotating shear system model. With this device it was possible to analyse the flow shear influencing washoff of a commercial fungicide formulation based on a copper oxychloride–metalaxyl mixture that was sprayed on a polypropylene surface. A factor plan with four variables, i.e. water speed and volume (both variables determining flow boundary stress in the shear device), formulation dosage and drying temperature, was set up to monitor colloid detachment. RESULTS: This experimental design, together with sorption experiments of metalaxyl on copper oxychloride, and the study of the dynamics of metalaxyl and copper oxychloride washoff, made it possible to prove that metalaxyl washoff from a polypropylene surface is controlled by transport in solution, whereas that of copper oxychloride occurs by particle detachment and transport of particles. Average losses for metalaxyl and copper oxychloride were, respectively, 29 and 50% of the quantity applied at the usual recommended dosage for crops. CONCLUSION: The key factors affecting losses were flow shear and the applied dosage. Empirical models using these factors provided good estimates of the percentage of fungicide loss. From the factor analysis, the main mechanism for metalaxyl loss induced by a shallow water flow is solubilisation, whereas copper loss is controlled by erosion of copper oxychloride particles. Copyright © 2009 Society of Chemical Industry     

Expression of the tobacco β-1,3-glucanase gene, PR-2d, following induction of SAR with Peronospora tabacina

Systemic acquired resistance (SAR) is induced following inoculation of Peronospora tabacina sporangia into the stems of Nicotiana tabacum plants highly susceptible to the pathogen. Previous results have shown that accumulation of acidic β-1,3-glucanases (PR-2's) following induction of SAR by P. tabacina may contribute to resistance to P. tabacina. We showed that up-regulation of the PR-2 gene, PR-2d, following stem inoculation with P. tabacina, is associated with SAR. Studies using plants transformed with GUS constructs containing the full length promoter from PR-2d or promoter deletions, provided evidence that a previously characterized regulatory element that is involved in response to salicylic acid (SA), may be involved in regulation of PR-2d following induction of SAR with P. tabacina. This work provides evidence that regulation of PR-2 genes during P. tabacina-induced SAR may be similar to regulation of these genes during infection of N-gene tobacco by TMV or following exogenous application of SA, and provides further support for the role of SA in regulation of genes during P. tabacina-induced SAR.     

In vitro and In vivo Activity of Cyprodinil and Pyrimethanil on Botrytis cinerea Isolates Resistant to other Botryticides and Selection for Resistance to Pyrimethanil in a Greenhouse Population in Greece

No cross-resistance was observed between pyrimethanil or cyprodinil and the fungicides benomyl, iprodione or carbendazim + diethofencarb. In vitro, both anilinopyrimidine fungicides were effective against strains of Botrytis cinerea resistant to benzimidazoles and/or dicarboximides and against a wild type strain insensitive to diethofencarb (EC₅₀ values ranged between 0.03–0.19 and 0.006–0.054gml^–1 for pyrimethanil and cyprodinil, respectively). Preventive applications of anilinopyrimidines completely protected young cucumber plants and fruits that were inoculated with all strains of B. cinerea. The effectiveness of pyrimethanil against grey mould was studied in greenhouse grown tomatoes in relation to (a) the type of infection and the progress of the disease on different plant parts and (b) the response of the naturally occurring B. cinerea population to the selection pressure caused by eight successive applications of this fungicide. Pyrimethanil effectively controlled grey mould on leaves, fruits and stems but did not significantly reduce the number of dead plants and fruits with 'ghost spot' symptoms. The selection pressure caused by the consecutive applications of pyrimethanil resulted in reduction of its effectiveness on leaves that became apparent after the sixth application. This was correlated with a shift of the B. cinerea population (not previously exposed to anilinopyrimidines) towards reduced sensitivity, probably due to the development of a low level of resistance (R _L = 7.7). Pyrimethanil delayed the onset of the disease but it did not reduce the infection rate.