A Reliable Method for Testing the Sensitivity of Botryotinia fuckeliana to Anilinopyrimidines In Vitro

Cyprodinil is a representative of the new class of broad-spectrum anilinopyrimidine fungicides. The effect of cyprodinil on mycelial growth of Botryotinia fuckeliana on solid agar medium depends on the composition of the medium and on the age of the mycelium to be used for the bioassay. An in-vitro method was developed to study the sensitivity distribution to cyprodinil in two wild-type populations of B. fuckeliana from fruits of strawberry with grey mould. To validate the applicability of the method, sensitivities to cyprodinil, mepanipyrim and pyrimethanil were monitored in populations of B. fuckeliana from grapes with grey mould from different vineyards, including one trial vineyard where reduced performance of cyprodinil had been encountered. The monitoring procedure was based on the inhibition of the mycelial growth on a synthetic medium containing L -asparagine (asp-agar) amended with the active ingredients. The mycelium was grown on asp-agar discs, starting from a spore suspension, for 17 h prior to the beginning of the test. This procedure proved to be efficient. The two wild-type populations from different sampling sites showed similar distributions of the sensitivity to cyprodinil. Some isolates from the trial site with reduced performance of anilinopyrimidines showed reduced sensitivities to cyprodinil, mepanipyrim and pyrimethanil, demonstrating cross-resistance between these anilinopyrimidines.     

Characterization of the bound residues of the fungicide cyprodinil formed in plant cell suspension cultures of wheat

The non-extractable residues of the fungicide cyprodinil formed in heterotrophic cell suspension cultures of wheat were studied by application of [2-pyrimidyl-14C] or [2-pyrimidyl-13C]cyprodinil. The main objective was to examine whether solid-state and liquid 13C NMR spectroscopy can be used to examine plant bound residues of pesticides. For 14C experiments, wheat suspensions grown on glucose as carbon source were treated with 10 mg litre(-1) of 14C-cyprodinil. After incubation for 12 days, 20% of applied 14C was detected as non-extractable residues. The cell debris were treated with 0.1 M HCl (reflux), 1.0 M HCl (reflux), buffer, or 2 M NaOH (50 degrees C); Bj?rkman lignin and acidolysis lignin fractions were also prepared from the debris. Radioactivity liberated and solubilized by these procedures was examined by thin-layer chromatography and high-performance liquid chromatography. The results showed that cyprodinil and primary metabolites contributed to the fungicide's bound residues. Most of the residues (12% of applied 14C) remained associated with polar or polymeric/oligomeric endogenous cell materials in a stable manner. For the study with 13C-cyprodinil, wheat suspensions were cultivated on 13C-depleted glucose for four growth cycles, resulting in maximum 13C depletion of the natural cell components to about 0.10%. During the fourth cycle, 13C-labelled cyprodinil was applied, and cells were incubated (12 days). Cell debris was prepared and examined by solid-state 13C NMR spectroscopy. Debris was then treated as described above in the 14C experiment. Solubilized fractions were analyzed by liquid 13C NMR spectroscopy. However, none of the 13C NMR spectra recorded gave utilizable or unambiguous results, and all exhibited large inconsistencies, especially concerning the data from the conventional 14C experiment.     

嘧菌环胺和缬霉威复配对5种作物病原菌的室内联合毒力

采用室内菌丝生长速率法,以草莓枯萎病菌、西瓜炭疽病菌、烟草猝倒病菌、葡萄霜霉病菌和月季灰霉病菌为作用目标,研究嘧菌环胺和缬霉威混配对5种病菌的联合毒力,为嘧菌环胺和缬霉威混配使用提供科学依据。结果显示,嘧菌环胺对5种病菌的有效中质量浓度(EC50)为0.19~1.92 mg/L,缬霉威对5种病菌的EC50为0.23~1.02 mg/L;当嘧菌环胺和缬霉威的混配比例为15∶1时,对草莓枯萎病菌的共毒系数最大,为217.70;混配比例为8∶1时,对西瓜炭疽病菌的共毒系数最大,为203.59;混配比例为3∶1时,对烟草猝倒病菌的共毒系数最大,为196.29;混配比例为1∶6时,对葡萄霜霉病菌的共毒系数最大,为201.98;混配比例为9∶1时,对月季灰霉病菌的共毒系数最大,为202.56。表明嘧菌环胺和缬霉威按适当比例混配对5种病原菌具有明显的增效作用。     

Chemical and biological control of Sclerotinia sclerotiorum in witloof chicory culture

BACKGROUND: Sclerotinia sclerotiorum (Lib.) de Bary is a major pathogen of witloof chicory. For lack of authorised field treatment, post‐harvest sprays with dicarboximide fungicides have been standard practice since the 1970s to prevent root rot and chicory heart decay during the forcing phase. However, the registration of procymidone and vinclozolin has been withdrawn in Europe. The development of organic agriculture and the necessity to reduce fungicide applications in conventional agriculture prompted an assessment of the efficacy of new fungicides and the use of the mycoparasite Coniothyrium minitans (Campbell). RESULTS: A mixture of the fungicides fludioxonil and cyprodinil (Switch^®) applied on chicory roots achieved a very good control of S. sclerotiorum (up to 95%). The use of C. minitans limited root infection, both when applied in the field (50–65% efficacy) and before the forcing period (post‐harvest treatment up to 80%). CONCLUSION: In organic agriculture, two treatments with C. minitans (in field and later at the forcing period) could improve protection against S. sclerotiorum. In conventional agriculture, after the field biological treatment, a post‐harvest chemical treatment could be applied. The addition of other prophylactic methods could lead to a high level of performance in practice against decay caused by S. sclerotiorum. Copyright © 2010 Society of Chemical Industry     

Pre- and post-inoculation activity of a protectant and a systemic fungicide for control of anthracnose fruit rot of strawberry under different wetness durations

A protectant fungicide (Captan, a.i. captan) and a systemic fungicide (Switch, a.i. fludioxonil + cyprodinil) were evaluated as pre- and post-inoculation applications for control of anthracnose fruit rot (AFR), caused by Colletotrichum acutatum, under a short (6 or 8 h) or long (18 or 24 h) wetting period. Evaluations were conducted for two seasons in Maryland and for two seasons in Florida. Both Captan and Switch were very effective for control of AFR when applied prior to inoculation, but control was more effective under the shorter wetting period. Switch was as effective when applied 4, 8, or 24 h post-inoculation as when applied before inoculation, but control was better under the short wetting period. Captan was effective when applied 4 or 8 h after inoculation under the short wetting period, but was ineffective at 24 h post-inoculation. Post-inoculation sprays of Captan were ineffective at any time under the long wetting period. The post-infection activity of Switch allows greater flexibility for managing AFR when fungicide applications are scheduled based on weather-based decision-support systems.     

嘧菌环胺原药高效液相色谱分析方法研究

本文采用高效液相色谱法,以甲醇+磷酸溶液为流动相,使用以ZORBAX Extend-C18、5μm为填料的不锈钢柱和二极管阵列检测器,在270nm波长下对嘧菌环胺原药进行分离和定量分析。结果表明,该分析方法的线性相关系数为0.999 7;标准偏差为0.19;变异系数为0.19%;平均回收率为100.1%。     

Resistance Risk Assessment of Cereal Eyespot, Tapesia Yallundae and Tapesia Acuformis, to the Anilinopyrimidine Fungicide, Cyprodinil

Eyespot pathogens, Tapesia yallundae and Tapesia acuformis, were isolated from two trial sites in the UK over several years. Both sites were treated with 2 applications per year of cyprodinil (a new anilinopyrimidine fungicide), prochloraz and a mixture of cyprodinil with prochloraz. One trial site was exposed to cyprodinil for 3 years, and the second for a total of 11 years, including 5 years before the trial was initiated. Control of eyespot and sensitivity to cyprodinil were monitored. During the first 3 years of the trial, disease control with all fungicide treatments ranged from 43% to 82%. At the site, where the trial was extended for a further 3 years, control then began to decline but no practical resistance was detected. The decline in control by both fungicides suggests that factors other than reduced sensitivity might be involved. Field isolates of both T. yallundae and T. acuformis with reduced sensitivity to cyprodinil were found predominantly in plots treated with cyprodinil. A reduction in sensitivity to cyprodinil was identified in the population from cyprodinil-treated plots in two years out of six, and in the population from mixture plots in the final year. No obvious trends could be identified and in-vivo studies showed control of most isolates with reduced sensitivity could be regained by increasing the dose to one tenth of the recommended field rate. Analysis of progeny from sexual crosses involving a sensitive isolate and a field isolate with an ED₅₀ value higher than the baseline sensitivity range indicated that a single gene controlled the reduction in sensitivity to cyprodinil in one T. yallundae isolate. There is clearly a resistance risk in eyespot to cyprodinil. The reduction in sensitivity is monogenic in inheritance and at a significant level in some isolates, but any shift in sensitivity in field populations has so far been gradual.     

The sex-specific sulfation of the major metabolite of the novel fungicide cyprodinil in the rat

The metabolism of cyprodinil, a novel broad-spectrum fungicide, was investigated in rats. After single oral administration of 0.5 or 100 mg kg⁻¹ body weight, [phenyl-U-¹⁴C]cyprodinil was rapidly eliminated, principally in the urine. The metabolite pattern in urine exhibited a significant sex-related difference with respect to the major metabolite. Males and females both produced a dihydroxy metabolite, N-4-(hydroxyphenyl)-4-cyclopropyl-5-hydroxy-6-methylpyrimidin-2-ylamine. Female rats conjugated this metabolite with sulfate exclusively at the 5-hydroxypyrimidinyl moiety, while males formed equal amounts of the monosulfate and a disulfate conjugate. The sex dimorphism in the conjugation reaction indicates the involvement of a sex-specific sulfotransferase that catalyzed the transfer of the second sulfate group.     

Evolution of fungicide resistance in the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis in France

Field isolates of the cereal eyespot pathogen can be divided into two groups which are now considered as two species: Tapesia yallundae and Tapesia acuformis. In both species the first case of acquired resistance was observed with benzimidazole fungicides in the early 1980s. At the same time, a number of sterol C-14 demethylation inhibitors (DMIs), such as the imidazole prochloraz and several triazoles, including flusilazole, were introduced. Surprisingly T. acuformis appeared intrinsically resistant to the triazole derivatives in comparison to T. yallundae, but both species were sensitive to prochloraz. The intensive use of these DMIs led to the development of acquired resistance towards triazoles in T. yallundae and towards prochloraz in T. acuformis. Today all the strains in both species appear equally sensitive to the anilinopyrimidine cyprodinil. ©1997 SCI     

嘧菌环胺在草莓上的残留降解行为研究

为评价嘧菌环胺在设施栽培草莓上使用后的残留行为及环境安全性,依据《农药残留试验准则》,采用田间试验方法,研究分析了设施栽培条件下嘧菌环胺在草莓上的消解动态和最终残留情况,并以残留分析结果为依据,对实际生产上的用药情况进行调整,探索最适安全间隔期。结果表明,以800倍液施药1次,嘧菌环胺在设施栽培草莓上的降解半衰期为3.180天;随施药浓度和次数的增加,嘧菌环胺残留量和膳食风险会加大,因此,安全间隔期应适当延长。