azoxystrobin and trifloxystrobin

The use of foliar fungicides on field corn has increased greatly over the past 5 years in the United States in an attempt to increase yields, despite limited evidence that use of the fungicides is consistently profitable. To assess the value of using fungicides in grain corn production, random-effects meta-analyses were performed on results from foliar fungicide experiments conducted during 2002 to 2009 in 14 states across the United States to determine the mean yield response to the fungicides azoxystrobin, pyraclostrobin, propiconazole + trifloxystrobin, and propiconazole + azoxystrobin. For all fungicides, the yield difference between treated and nontreated plots was highly variable among studies. All four fungicides resulted in a significant mean yield increase relative to the nontreated plots (P < 0.05). Mean yield difference was highest for propiconazole + trifloxystrobin (390 kg/ha), followed by propiconazole + azoxystrobin (331 kg/ha) and pyraclostrobin (256 kg/ha), and lowest for azoxystrobin (230 kg/ha). Baseline yield (mean yield in the nontreated plots) had a significant effect on yield for propiconazole + azoxystrobin (P < 0.05), whereas baseline foliar disease severity (mean severity in the nontreated plots) significantly affected the yield response to pyraclostrobin, propiconazole + trifloxystrobin, and propiconazole + azoxystrobin but not to azoxystrobin. Mean yield difference was generally higher in the lowest yield and higher disease severity categories than in the highest yield and lower disease categories. The probability of failing to recover the fungicide application cost (p(loss)) also was estimated for a range of grain corn prices and application costs. At the 10-year average corn grain price of $0.12/kg ($2.97/bushel) and application costs of $40 to 95/ha, p(loss) for disease severity <5% was 0.55 to 0.98 for pyraclostrobin, 0.62 to 0.93 for propiconazole + trifloxystrobin, 0.58 to 0.89 for propiconazole + azoxystrobin, and 0.91 to 0.99 for azoxystrobin. When disease severity was >5%, the corresponding probabilities were 0.36 to 95, 0.25 to 0.69, 0.25 to 0.64, and 0.37 to 0.98 for the four fungicides. In conclusion, the high p(loss) values found in most scenarios suggest that the use of these foliar fungicides is unlikely to be profitable when foliar disease severity is low and yield expectation is high.

Topics: Acetates; Carbamates; Edible Grain; Fungicides, Industrial; Imines; Methacrylates; Plant Diseases; Plant Leaves; Pyrazoles; Pyrimidines; Risk Factors; Strobilurins; Triazoles; United States; Zea mays

Despite the widespread application of strobilurin fungicides (SFs) in agriculture, little is known about their distribution and bioaccumulation in aquatic ecosystems. In this study, the concentrations of 12 SFs and two of their metabolites were determined in abiotic (water and sediment; n = 83) and biotic (plant, algae, zooplankton, and fish; n = 123) samples collected from a subtropical freshwater ecosystem, namely, Dongjiang River wetland, in southern China. Among the 12 SFs measured, azoxystrobin (AZ) was the major fungicide found in surface water (median: 2.20 ng/L) and sediment (0.064 ng/g dry wt.). Azoxystrobin acid (AZ-acid), a metabolite of AZ, was the major analyte in the plant samples and had a median concentration at 0.36 ng/g dry wt. In algae and zooplankton, (Z)-metominostrobin was the predominant fungicide and had median concentrations of 3.52 and 5.55 ng/g dry wt., respectively. In fish muscle, dimoxystrobin (DIMO) was the major SF and had a median concentration of 0.47 ng/g dry wt. The bioconcentration factor (BCF) values of AZ-acid, trifloxystrobin (TFS), and pyraclostrobin (PYR) in algae and zooplankton and AZ-acid, PYR, TFS, TFS-acid, picoxystrobin, and DIMO in fish muscle exceeded 1000 L/kg (algae, zooplankton, and fish concentrations were expressed on a dry weight basis), suggesting that these fungicides can accumulate in biota. A positive association between log BCFs of SFs in fish and logKow of SFs and a negative correlation between log BCFs and the log solubility index were observed. Additionally, the risk quotient (RQ) was calculated to evaluate the potential ecotoxicological risk of SFs to different aquatic organisms (algae, zooplankton, and fish). The PYR and DIMO concentrations at 19 sampling sites had RQ values >0.1, indicating moderate ecotoxicological risks to aquatic organisms. This study is the first to document the widespread occurrence of SFs and their metabolites in aquatic ecosystems and to elucidate the bioaccumulation potential of SFs in aquatic organisms.

Topics: Animals; Aquatic Organisms; Bioaccumulation; China; Ecosystem; Environmental Monitoring; Fishes; Fungicides, Industrial; Rivers; Strobilurins; Water; Water Pollutants, Chemical; Zooplankton

Strobilurin fungicides are quinone outside inhibitors (QoI) used to treat fungal pathogens for agricultural and residential use. Here, we compared the potential for neurotoxicity of the widely used strobilurins, azoxystrobin (AZS) and trifloxystrobin (TFS), in differentiated human SH-SY5Y cells. Fungicides did not include cytotoxicity up to 200 µM but both induced loss of cell viability at 48 h, with TFS showing slightly higher toxicity that AZS. Caspase 3/7 activity was induced in SH-SY5Y cells by both fungicides at 48 h (50 µM for AZS and 25 µM for TFS). ATP levels were reduced following a 24-hour exposure to > 25 µM AZS and > 6.25 µM TFS and both fungicides rapidly impaired oxidative respiration (~12.5 µM for AZS and ~3.125 µM TFS) and decreased oligomycin-induced ATP production, maximal respiration, and mitochondrial spare capacity. AZS at 100 µM showed a continual impairment of mitochondrial membrane potential (MMP) between 4 and 48 h while TFS at > 50 µM decreased MMP at 24 h. Taken together, TFS exerted higher mitochondrial toxicity at lower concentrations compared to AZS in SH-SY5Y cells. To discern toxicity mechanisms of strobilurin fungicides, lipidomics was conducted in SH-SY5Y cells following exposure to 6.25 µM and 25 µM AZS, and a total of 1595 lipids were detected, representing 49 different lipid classes. Lipid classes with the largest proportion of lipids detected in SH-SY5Y cells included triglycerides (17%), phosphatidylethanolamines (8%), ether-linked triglycerides (8%), phosphatidylcholines (7%), ether-linked phosphatidylethanolamines (6%), and diacylglycerols (5%). Together, these 5 lipid classes accounted for over 50% of the total lipids measured in SH-SY5Y cells. Lipids that were increased by AZS included acyl carnitine, which plays a role in long chain fatty acid utilization for mitochondrial β-oxidation, as well as non-modified, ether linked, and oxidized triacylglycerols, suggesting compensatory upregulation of triglyceride biosynthesis. The ceramide HexCer-NS, linked to neurodegenerative diseases, was decreased in abundance following AZS exposure. In summary, strobilurin fungicides rapidly inhibit mitochondrial oxidative respiration and alter the abundance of several lipids in neuronal cells, relevant for understanding environmental exposure risks related to their neurotoxicity.

Topics: Acetates; Adenosine Triphosphate; Cell Line, Tumor; Ethers; Fungicides, Industrial; Humans; Imines; Lipidomics; Membrane Potential, Mitochondrial; Neuroblastoma; Neurotoxicity Syndromes; Phosphatidylethanolamines; Pyrimidines; Strobilurins; Triglycerides

Strobilurin fungicides are used primarily in fruits and vegetables, but recently, a patent was issued for one strobilurin fungicide, azoxystrobin, in mold-resistant wallboard. This raises concerns about the potential presence of these chemicals in house dust and potential exposure indoors, particularly in young children. Furthermore, recent toxicological studies have suggested that strobilurins may cause neurotoxicity. Currently, it is not clear whether or not azoxystrobin applications in wallboard lead to exposures in the indoor environments. The purpose of this study was to determine if azoxystrobin, and related strobilurins, could be detected in house dust. We also sought to characterize the concentrations of azoxystrobin in new wallboard samples. To support this study, we collected and analyzed 16 new dry wall samples intentionally marketed for use in bathrooms to inhibit mold. We then analyzed 188 house dust samples collected from North Carolina homes in 2014-2016 for azoxystrobin and related strobilurins, including pyraclostrobin, trifloxystrobin, and fluoxastrobin using liquid chromatography tandem mass spectrometry. Detection frequencies for azoxystrobin, pyraclostrobin, trifloxystrobin, and fluoxastrobin ranged from 34-87%, with azoxystrobin being detected most frequently and at the highest concentrations (geometric mean = 3.5 ng/g; maximum = 10,590 ng/g). Azoxystrobin was also detected in mold-resistant wallboard samples, primarily in the paper covering where it was found at concentrations up to 88.5 µg/g. Cumulatively, these results suggest that fungicides present in wallboard may be migrating to the indoor environment, leading to exposure in the residences that would constitute a separate exposure pathway independent of dietary exposures.

Topics: Acetates; Air Pollution, Indoor; Chromatography, Liquid; Construction Industry; Dust; Fungicides, Industrial; Imines; North Carolina; Pyrimidines; Strobilurins

Chromoblastomycosis (CBM) is a chronic subcutaneous mycosis caused by traumatic implantation of many species of black fungi. Due to the refractoriness of some cases and common recurrence of CBM, a more effective and less time-consuming treatment is mandatory. The aim of this study was to identify compounds with in vitro antifungal activity in the Pathogen Box® compound collection against different CBM agents. Synergism of these compounds with drugs currently used to treat CBM was also assessed. An initial screening of the drugs present in this collection at 1 μM was performed with a Fonsecaea pedrosoi clinical strain according to the EUCAST protocol. The compounds with activity against this fungus were also tested against other seven etiologic agents of CBM (Cladophialophora carrionii, Phialophora verrucosa, Exophiala jeanselmei, Exophiala dermatitidis, Fonsecaea monophora, Fonsecaea nubica, and Rhinocladiella similis) at concentrations ranging from 0.039 to 10 μM. The analysis of potential synergism of these compounds with itraconazole and terbinafine was performed by the checkerboard method. Eight compounds inhibited more than 60% of the F. pedrosoi growth: difenoconazole, bitertanol, iodoquinol, azoxystrobin, MMV688179, MMV021013, trifloxystrobin, and auranofin. Iodoquinol produced the lowest MIC values (1.25-2.5 μM) and MMV688179 showed MICs that were higher than all compounds tested (5 - >10 μM). When auranofin and itraconazole were tested in combination, a synergistic interaction (FICI = 0.37) was observed against the C. carrionii isolate. Toxicity analysis revealed that MMV021013 showed high selectivity indices (SI ≥ 10) against the fungi tested. In summary, auranofin, iodoquinol, and MMV021013 were identified as promising compounds to be tested in CBM models of infection.

Topics: Acetates; Antifungal Agents; Ascomycota; Auranofin; Biphenyl Compounds; Chromoblastomycosis; Dioxolanes; Drug Synergism; Exophiala; Fungi; Humans; Imines; Iodoquinol; Pyrimidines; Strobilurins; Triazoles

The long-term decline of monarch butterflies has been attributed to loss of their milkweed (Asclepias sp.) host-plants after the introduction of herbicide-tolerant crops. However, recent studies report pesticide residues on milkweed leaves that could act as a contributing factor when ingested as part of their larval diet. In this study, we exposed monarch larvae to six pesticides (insecticide: clothianidin; herbicides: atrazine, S-metolachlor; fungicides: azoxystrobin, pyraclostrobin, trifloxystrobin) on their primary host-plant, A. syriaca. Each was tested at mean and maximum levels reported from published analyses of milkweeds bordering cropland and thus represent field-relevant concentrations. Monarch lethal and sub-lethal responses were tracked over their complete development, from early instar larvae to adult death. Overall, we found no impact of any pesticide on immature development time and relatively weak effects on larval herbivory or survival to adulthood. Comparatively stronger effects were detected for adult performance; namely, a 12.5% reduction in wing length in response to the fungicides azoxystrobin and trifloxystrobin. These data collectively suggest that monarch responses to host-plant pesticides are largely sublethal and more pronounced in the adult stage, despite exposure only as larvae. This outcome has important implications for risk assessment and the migratory success of monarchs in North America.

Topics: Acetamides; Acetates; Animal Migration; Animals; Asclepias; Atrazine; Butterflies; Ecosystem; Fungicides, Industrial; Herbicides; Herbivory; Imines; Larva; Pesticides; Population Dynamics; Pyrimidines; Risk Assessment; Strobilurins

A novel microextraction technique combining magnetic solid-phase microextraction (MSPME) with ionic liquid dispersive liquid-liquid microextraction (IL-DLLME) to determine four fungicides is presented in this work for the first time. The main factors affecting the extraction efficiency were optimized by the one-factor-at-a-time approach and the impacts of these factors were studied by an orthogonal design. Without tedious clean-up procedure, analytes were extracted from the sample to the adsorbent and organic solvent and then desorbed in acetonitrile prior to chromatographic analysis. Under the optimum conditions, good linearity and high enrichment factors were obtained for all analytes, with correlation coefficients ranging from 0.9998 to 1.0000 and enrichment factors ranging 135 and 159 folds. The recoveries for proposed approach were between 98% and 115%, the limits of detection were between 0.02 and 0.04 μg L(-1) and the RSDs changed from 2.96 to 4.16. The method was successfully applied in the analysis of four fungicides (azoxystrobin, chlorothalonil, cyprodinil and trifloxystrobin) in environmental water samples. The recoveries for the real water samples ranged between 81% and 109%. The procedure proved to be a time-saving, environmentally friendly, and efficient analytical technique.

Topics: Acetates; Acetonitriles; beta-Cyclodextrins; Chromatography, High Pressure Liquid; Fungicides, Industrial; Imines; Ionic Liquids; Liquid Phase Microextraction; Magnesium Compounds; Magnetic Phenomena; Methacrylates; Nitriles; Pyrimidines; Silicon Compounds; Solid Phase Microextraction; Solvents; Strobilurins; Water Pollutants, Chemical

Calonectria (formerly Cylindrocladium) infection of pot azalea (Rhododendron simsii Planch) is an important disease problem in which usually one or two of the four plants per pot show progressing leaf and especially stem lesions, leading to mortality of the respective plant and rendering the pot unmarketable. This may occur in a later stage of the growing season, leading to significant commercial losses. The main objective of this study was to test a range of fungicides for their efficacy against this pathogen. To test the fungicides, a bioassay was first developed in which mycelium and conidiospores of the pathogen were produced on Potato Dextrose Agar, blended in water, and dilutions of the resulting suspension inoculated at the base of 11-week-old cuttings three weeks after they had been trimmed. Disease progression was monitored up to 7 weeks post inoculation and a disease index on a scale of 0 to 3 was established. In the actual efficacy trial, the following fungicides (with corresponding active ingredient(s)) were tested as preventive treatments: Topsin M 70 WG (thiophanate-methyl), Sporgon (prochloraz), Signum (boscalid+pyraclostrobin), Switch (cyprodinyl+fludioxonil), Flint 50WG (trifloxystrobin), Ortiva Top (azoxystrobin+difenoconazole) and Fungaflor (imazalil). Disease expression started after about 2 weeks, increased approximately 1 index level, and leveled off 5 weeks after inoculation. The best control was observed with Sporgon, Ortiva Top and Signum. Switch produced intermediate effects and insufficient control was observed with Topsin, Flint and Fungaflor. These results explain why specific standard fungicide treatments, such as those with Topsin, fail to control the disease, while they can be effective against a different Calonectria species such as C. pseudonaviculata, the cause of boxwood blight.

Topics: Acetates; Carbamates; Dioxolanes; Fungicides, Industrial; Hypocreales; Imidazoles; Imines; Methacrylates; Plant Diseases; Pyrazoles; Pyrimidines; Rhododendron; Strobilurins; Triazoles

The first strobilurin fungicides were introduced in 1996, and have since been used in a vast array of disease/plant systems worldwide. The strobilurins now consist of 16 compounds and represent the 2nd most important fungicide group worldwide with 15% of the total fungicide market share. Strobilurins are moderately persistent in soil, and some degradation products (e.g. azoxystrobin acid) have been detected as contaminants of freshwater systems. Little is currently known about the transformation processes involved in the biodegradation of strobilurins or the microbial groups involved. Using sequential soil and liquid culture enrichments, we isolated two bacterial strains which were able to degrade the most widely used strobilurin, azoxystrobin, when supplied as a sole carbon source. 16S rRNA showed that the strains showed homology to Cupriavidus sp. and Rhodanobacter sp. Both isolated strains were also able to degrade the related strobilurin compounds trifloxystrobin, pyraclostrobin, and kresoxim-methyl. An additional nitrogen source was required for degradation to occur, but the addition of a further carbon source reduced compound degradation by approximately 50%. However, (14)C radiometric analysis showed that full mineralisation of azosxystrobin to (14)CO2 was negligible for both isolates. 16S rRNA T-RFLP analysis using both DNA and RNA extracts showed that degradation of azoxystrobin in soil was associated with shifts in bacterial community structure. However, the phylotypes which proliferated during degradation could not be attributed to the isolated degraders.

Topics: Acetates; Bacteria; Biodegradation, Environmental; Fungicides, Industrial; Imines; Methacrylates; Phenylacetates; Polymorphism, Restriction Fragment Length; Pyrimidines; RNA, Ribosomal, 16S; Soil; Soil Microbiology; Soil Pollutants; Strobilurins

The simultaneous determination of four strobilurin fungicides (picoxystrobin, kresoxim-methyl, trifloxystrobin, and azoxystrobin) in cotton seed by combining acetonitrile extraction and dispersive liquid-liquid microextraction was developed prior to GC with electron capture detection. Several factors, including the type and volume of the extraction and dispersive solvents, extraction condition and time, and salt addition, were optimized. The analytes were extracted with acetonitrile from cotton seed and the clean-up was carried out by primary secondary amine. Afterwards, 60 μL of n-hexane/toluene (1:1, v/v) with a lower density than water was mixed with 1 mL of the acetonitrile extract, then the mixture was injected into 7 mL of distilled water. A 0.1 mL pipette was used to collect a few microliters of n-hexane/toluene from the top of the aqueous solution. The enrichment factors of the analytes ranged from 36 to 67. The LODs were in the range of 0.1 × 10(-3) -2 × 10(-3) mg/kg. The relative recoveries varied from 87.7 to 95.2% with RSDs of 4.1-8.5% for the four fungicides. The good performance of the method, compared with the conventional pretreatments, has demonstrated it is suitable for determining low concentrations of strobilurin fungicide residues in cotton seed.

Topics: Acetates; Acetonitriles; Acrylates; Chromatography, Gas; Fungicides, Industrial; Gossypium; Imines; Liquid Phase Microextraction; Methacrylates; Phenylacetates; Pyridines; Pyrimidines; Seeds; Strobilurins

An LC-MS/MS method was developed for the simultaneously determination of seven strobilurin fungicide residues in Chinese herbs. The strobilurin fungicides include Z-metominostrobin, kresoxim-methyl, dimoxystrobin, picoxystrobin, pyraclostrobin, azoxystrobin and trifloxystrobin. The sample was extracted with ethyl acetate and cleaned-up by an amino SPE column. The seven strobilurin fungicide residues were separated on a C18 column with gradient elution of 1.0 per thousand formic acid and methanol as mobile phases, and detected by ESI-MS in positive ion and selective reaction monitoring (SRM) mode. External standard method was used to the quantification with good linear relationships (r > or = 0. 996). The LOQs were 2 micro g/kg for dimoxystrobin, picoxystrobin and trifloxystrobin, 4 mciro g/kg for pyraclostrobin and azoxystrobin, 10 micro g/kg for Z-metominostrobin and kresoxim-methyl. The recoveries were from 60.4% to 110% with the RSDs between 1.2% and 17%. The developed method is suitable for the determination and confirmation of the seven strobilurin fungicide residues in the three of Eight Zhes ( Ophiopogon japonicus (Thunb.), Scrophularia ningpoensis Hemsl. and Corydalis yanhusuo W T Wang).

Topics: Acetates; Acrylates; Carbamates; Chromatography, Liquid; Drug Contamination; Drug Residues; Drugs, Chinese Herbal; Fungicides, Industrial; Imines; Methacrylates; Phenylacetates; Pyrazoles; Pyridines; Pyrimidines; Strobilurins; Tandem Mass Spectrometry

The strobilurins are used widely in the world as effective fungicidal agents to control Asian soybean rust. In this study, the early life stage of grass carp (Ctenopharyngodon idella), which is one of the most important aquaculture species in China, was chosen to measure the acute toxicity of three common strobilurin-derived fungicides (trifloxystrobin (TFS), azoxystrobin (AZ) and kresoxim-methyl (KM)). As endpoints, normal developmental parameters (lethal concentration (LC₅₀) and average heart rate), expression of relative genes, and three antioxidant enzyme activities in the developing juveniles were recorded during a 48 h exposure. The results revealed that values of LC₅₀ were TFS 0.051 (0.046-0.058) mg L⁻¹, AZ 0.549 (0.419-0.771) mg L⁻¹ and KM 0.338 (0.284-0.407) mg L⁻¹ for juveniles. For the potential toxicity mechanisms, these three fungicides increased catalase (CAT) and peroxidase (POD) activity and decreased superoxide dismutase (SOD) activity, significantly inhibited expressions of three growth-related genes (IGF-1, IGF-2 and GHR) and two energy-related-genes (CCK and PYY), and caused pronounced up-regulation a stress-gene (HSP70). The present study demonstrated potential toxic effects of TFS, AZ and KM on the early development of C. idella. Overall, three strobilurins (TFS, AZ and KM) might cause serious damages to the aquatic species; therefore, their pollution supervision in water ecological environment should be strengthened.

Topics: Acetates; Animals; Carps; Catalase; China; Fungicides, Industrial; Gene Expression; HSP72 Heat-Shock Proteins; Imines; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Methacrylates; Peroxidase; Phenylacetates; Pyrimidines; Receptors, Somatotropin; RNA, Messenger; Strobilurins; Superoxide Dismutase

Strobilurin fungicides are a leading class of antifungal chemicals used today in agricultural applications. Although degradation of some strobilurin fungicides has been assessed in plant residues, little information has appeared in the literature concerning the rates of metabolism of these fungicides in plants. In this study, we explored plant metabolism of three strobilurin fungicides, azoxystrobin, kresoxim-methyl, and trifloxystrobin, using wheat cell suspension cultures. Trifloxystrobin and kresoxim-methyl were completely metabolized within 24 h, whereas the metabolism of azoxystrobin was relatively slow with half-lives up to 48 h depending on specific experimental conditions. Metabolic rates of these fungicides were affected by the amounts of compound and cells added to the media. Structural analysis of metabolites of trifloxystrobin and kresoxim-methyl by high-resolution mass spectrometry (HRMS) and nuclear magnetic resonance spectroscopy (NMR) indicated that trifloxystrobin was first demethylated followed by subsequent hydroxylation, whereas kresoxim-methyl was largely demethylated. In contrast, a number of minor metabolites of azoxystrobin were present suggesting a differential metabolism of strobilurins by wheat cells.

Topics: Acetates; Cells, Cultured; Fatty Acids, Unsaturated; Fungicides, Industrial; Hydrolysis; Imines; Kinetics; Magnetic Resonance Spectroscopy; Mass Spectrometry; Methacrylates; Phenylacetates; Pyrimidines; Strobilurins; Triticum

QoI fungicides, inhibitors of mitochondrial respiration, are considered to be at high risk of resistance development. In several phytopathogenic fungi, resistance is caused by mutations (most frequently G143A) in the mitochondrial cytochrome b (cytb) gene. The genetic and molecular basis of QoI resistance were investigated in laboratory and field mutants of Botryotinia fuckeliana (de Bary) Whetz. exhibiting in vitro reduced sensitivity to trifloxystrobin.. B. fuckeliana mutants highly resistant to trifloxystrobin were obtained in the laboratory by spontaneous mutations in wild-type strains, or from naturally infected plants on a medium amended with 1-3 mg L(-1) trifloxystrobin and 2 mM salicylhydroxamic acid, an inhibitor of alternative oxidase. No point mutations were detected, either in the complete nucleotide sequences of the cytb gene or in those of the aox and Rieske protein genes of laboratory mutants, whereas all field mutants carried the G143A mutation in the mitochondrial cytb gene. QoI resistance was always maternally inherited in ascospore progeny of sexual crosses of field mutants with sensitive reference strains.. The G143A mutation in cytb gene is confirmed to be responsible for field resistance to QoIs in B. fuckeliana. Maternal inheritance of resistance to QoIs in progeny of sexual crosses confirmed that it is caused by extranuclear genetic determinants. In laboratory mutants the heteroplasmic state of mutated mitochondria could likely hamper the G143A detection, otherwise other gene(s) underlying different mechanisms of resistance could be involved.

Topics: Acetates; Botrytis; Cytochromes b; Drug Resistance, Fungal; Enzyme Inhibitors; Fungal Proteins; Fungicides, Industrial; Imines; Methacrylates; Mutation, Missense; Strobilurins; Vitis

The Quinone outside Inhibitors (QoI) are one of the most important and recent fungicide groups used in viticulture and also allowed by Integrated Pest Management. Azoxystrobin, kresoxim-methyl and trifloxystrobin are the main active ingredients for treating downy and powdery mildews that can be present in grapes and wines. In this paper, a method is reported for the analysis of these three QoI-fungicides in grapes and wine. After liquid-liquid extraction and a clean-up on commercial silica cartridges, analysis was by isocratic HPLC with diode array detection (DAD) with a run time of 13 min. Confirmation was by solid-phase micro-extraction (SPME), followed by GC/MS determination. The main validation parameters for the three compounds in grapes and wine were a limit of detection up to 0.073 mg kg(-1), a precision not exceeding 10.0% and an average recovery of 93% +/- 38.

Topics: Acetates; Acrylates; Chromatography, High Pressure Liquid; Food Analysis; Food Contamination; Fungicides, Industrial; Imines; Methacrylates; Pesticide Residues; Phenylacetates; Pyrimidines; Reproducibility of Results; Strobilurins; Vitis; Wine

Qo-inhibiting fungicides act as respiration inhibitors by binding to the Qo center of cytochrome b. Sensitivities of fungi to Qo inhibitors can be influenced by the induction of alternative respiration or by mutational changes of the cytochrome b target site. Previous studies on both mechanisms in Magnaporthe grisea (Hebert) Barr were focused on the mycelial stage of the pathogen. The present study describes the expression and impact of both resistance mechanisms during the stage of conidia germination. In the absence of a host, alternative respiration provided a >500-fold rescue from azoxystrobin during the germination of conidia derived from four wild-type isolates of M. grisea. This rescue potential during conidia gemination was substantially more pronounced than for mycelial growth. However, the pronounced effectiveness of alternative respiration during conidia germination was not apparent when barley leaves were protected with azoxystrobin prior to inoculation with conidia. In a comparison of a wild-type strain and an alternative respiration-deficient mutant, azoxystrobin efficacies in suppressing symptom development differed by a factor of two, with full disease control achieved for both genotypes at 1 microg ml(-1) azoxystrobin. In contrast, conidia derived from two QoI-resistant target site mutants were highly resistant to azoxystrobin and trifloxystrobin and fully capable of infecting leaf surfaces protected with 10 microg ml(-1) of azoxystrobin. Both target-site mutants had emerged spontaneously in the presence of high azoxystrobin doses when residual mycelial growth was supported by alternative respiration. The effective silencing of alternative respiration in protective applications of Qo-inhibiting fungicides might constitute a strategy of slowing the emergence of highly resistant target site mutants.

Topics: Acetates; Acrylates; Cytochromes b; Dose-Response Relationship, Drug; Drug Interactions; Drug Resistance, Fungal; Fungicides, Industrial; Gene Expression; Germination; Hordeum; Imines; Magnaporthe; Methacrylates; Mutation; Oxygen Consumption; Plant Leaves; Pyrimidines; Spores; Strobilurins

Strobilurin fungicides have a broad spectrum activity against all major foliar pathogens of wheat. In addition to this extraordinary fungicidal activity side-effects have been reported which result in higher yields of cereals, e.g. the reduction of respiration, delayed leaf senescence, activation of nitrogen metabolism as well as increased tolerance against abiotic stress factors. In the vegetation period 2000/2001 field trials were carried out at three sites in North Rhine-Westphalia to study the effects of three strobilurin fungicides on the yield formation of six winter wheat varieties. The strobilurins were applied two times as the commercial products Stratego (trifloxystrobin + propiconazole), Amistar/Pronto Plus (azoxystrobin/spiroxamine + tebuconazole) and Juwel Top (kresoxim-methyl + epoxiconazole + fenpropimorph. Fungicide-treated plants were kept disease-free by an initial azole-application in GS 31 in order to exclude disease effects on physiological parameters relevant to yield formation. Photosynthetic electron transport of strobilurin-treated wheat, was improved as early as at GS 65 compared to azole-treated plants. Differences often increased with growth stage and were closely related to a delay in leaf senescence. A higher photosynthetic activity of strobilurin-treated plants was confirmed by gas exchange and chlorophyll fluorescence measurements under field conditions. The yield benefit of wheat from strobilurin treatments varied from 2% to 9% depending on an improved photosynthetic capacity due to a higher and/or prolonged activity. Neither yield potential nor disease susceptibility of the cultivar had an effect on the height of the extra yield which, in contrast was modified by location and wheat genotype.

Topics: Acetates; Acrylates; Azoles; Chlorophyll; Epoxy Compounds; Fungicides, Industrial; Germany; Imines; Methacrylates; Morpholines; Phenylacetates; Photosynthesis; Pyrimidines; Strobilurins; Triazoles; Triticum