azoxystrobin and kresoxim-methyl

A nonaqueous micellar electrokinetic capillary chromatography method with indirect LIF was developed for the determination of strobilurin fungicide residues in fruits and vegetables. Hydrophobic CdTe quantum dots (QDs) synthesized in aqueous phase were used as background fluorescent substance. The BGE solution, QD concentration, and separation voltage were optimized to obtain the best separation efficiency and the highest signal intensity. The optimal BGE solution consists of 40 mM phosphate, 120 mM sodium dodecyl sulfate, 15% v/v water and 15% v/v hydrophobic CdTe QDs in formamide, of which apparent pH is 9.5. The optimized separation voltage is controlled as 25 kV. The resultant detection limits of azoxystrobin, kresoxim-methyl, and pyraclostrobin are all 0.001 mg/kg, their linear dynamic ranges are 0.005-2.5 mg/kg, and the recoveries of the spiked samples are 81.7-96.1%, 86.5-95.7%, and 87.3-97.4%, respectively. This method has been proved to be sensitive enough to detect the aforementioned fungicides in fruits and vegetables at the maximum residue limits.

Topics: Chromatography, Micellar Electrokinetic Capillary; Fruit; Fungicides, Industrial; Humans; Hydrophobic and Hydrophilic Interactions; Lasers; Limit of Detection; Methacrylates; Pesticide Residues; Pyrimidines; Quantum Dots; Spectrometry, Fluorescence; Strobilurins; Vegetables

Azoxystrobin and kresoxim-methyl are strobilurin fungicides, and are effective in controlling many plant diseases, including Fusarium wilt. The mode of action of this kind of chemical is inhibition of respiration. This research investigated the sensitivities of Fusarium kyushuense to azoxystrobin and kresoxim-methyl, and to the alternative oxidase inhibitor salicylhydroxamic acid (SHAM). The Biolog FF MicroPlate is designed to examine substrate utilization and metabolic profiling of micro-organisms, and was used here to study the activity of azoxystrobin, kresoxim-methyl and SHAM against F. kyushuense. Results presented that azoxystrobin and kresoxim-methyl strongly inhibited conidial germination and mycelial growth of F. kyushuense, with EC50 values of 1.60 and 1.79μgml(-1), and 6.25 and 11.43μgml(-1), respectively; while not for SHAM. In the absence of fungicide, F. kyushuense was able to metabolize 91.6% of the tested carbon substrates, including 69 effectively and 18 moderately. SHAM did not inhibit carbon substrate utilization. Under the selective pressure of azoxystrobin and kresoxim-methyl during mycelial growth (up to 100μgml(-1)) and conidial germination (up to 10μgml(-1)), F. kyushuense was unable to metabolize many substrates in the Biolog FF MicroPlate; while especially for carbon substrates in glycolysis and tricarboxylic acid cycle, with notable exceptions such as β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, α-d-glucose-1-phosphate, d-saccharic acid and succinic acid in the mycelial growth stage, and β-hydroxybutyric acid, y-hydroxybutyric acid, α-ketoglutaric acid, tween-80, arbutin, dextrin, glycerol and glycogen in the conidial germination stage. This is a new finding for some effect of azoxystrobin and kresoxim-methyl on carbon substrate utilization related to glycolysis and tricarboxylic acid cycle and other carbons, and may lead to future applications of Biolog FF MicroPlate for metabolic effects of other fungicides and other fungi, as well as providing a carbon metabolic fingerprint of F. kyushuense that could be useful for identification.

Topics: Citric Acid Cycle; Fungicides, Industrial; Fusarium; Glycolysis; Methacrylates; Phenylacetates; Pyrimidines; Strobilurins

Phenamacril (JS399-19 with independent intellectual property developed by China), azoxystrobin, and kresoxim-methyl are strobilurin fungicide. Due to their broad spectrum and good control of most of known fungi, strobilurin fungicide has been widely used in agriculture management. Thus, it is important to evaluate their environmental behaviors particularly in soils and underground water. In this study, the sorption/desorption and mobility of strobilurin fungicides in three Chinese soils (Jiangxi red soil, Taihu paddy soil, and Northeast China black soil) were conducted using comprehensively analytic approaches including batch experiment and soil thin-layer chromatography. The strobilurin fungicides were hard to be adsorbed in Jiangxi red soil but had medium adsorption capability in Tanhu paddy soil and Northeast China black soil, while the desorption of three strobilurin fungicides ranked in the order of Jiangxi red soil > Taihu paddy soil > Northeast China black soil. Soil properties including soil organic matter (SOM), pH, and cationic exchange capacity (CEC) affected the adsorption/desorption of the fungicides. Azoxystrobin and kresoxim-methyl had weak mobility in the soils. JS399-19 was moderately mobile in Jiangxi red soil but was not easily moved in Taihu paddy soil and Northeast China black soil. Due to their weak mobility in soils, these strobilurin fungicides tended to remain in the soil phase but not to shift downward to underground water. As azoxystrobin and JS399-19 had a long retention period in soil, there may become persistent residues in the soil environment.

Topics: Adsorption; Agriculture; Amino Acids; China; Fungicides, Industrial; Methacrylates; Phenylacetates; Phenylpropionates; Pyrimidines; Soil; Soil Pollutants; Strobilurins; Water Pollutants, Chemical

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

A new assay of micellar electrokinetic capillary chromatography with sweeping was developed to determine azoxystrobin, kresoxim-methyl and pyraclostrobin in fruits and vegetables. The key factors affecting resolution and peak height were studied and the optimum conditions were obtained for separation and enrichment. The running buffer consisted of 40 mM borate, 25 mM sodium dodecyl sulfate and 15% acetonitrile, and its pH was adjusted to 8.4. The sample was injected for 677 nL and the separation voltage was 25 kV. Under the optimum conditions, the enrichment factors of azoxystrobin, kresoxim-methyl and pyraclostrobin were 861, 550 and 403; the linear dynamic ranges were all 0.01-5.0 mg/L; the limits of detection were 0.002, 0.001 and 0.002 mg/kg; the recoveries of spiked samples were 85.1-98.5%, 87.5-97.0% and 89.1-99.1%, respectively. The assay can meet the requirement of maximum residue limits for these three strobilurin fungicides, and has been applied for determining their residues in fruits and vegetables.

Topics: Acetonitriles; Carbamates; Chromatography, Micellar Electrokinetic Capillary; Fruit; Fungicides, Industrial; Hydrogen-Ion Concentration; Limit of Detection; Methacrylates; Pesticide Residues; Phenylacetates; Pyrazoles; Pyrimidines; Reproducibility of Results; Strobilurins; Vegetables

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

Although there is some evidence regarding the effect of solarization and biosolarization on pesticide degradation, information is still scarce. The aim of this study was to determine the effect of these disinfection techniques on the degradation of eight fungicides (azoxystrobin, kresoxin methyl, tebuconazole, hexaconazole, triadimenol, cyprodinil, pyrimethanil and fludioxonil) commonly used in pepper crops under greenhouse cultivation. Seventy-five 17-L pots filled with clay-loam soil were placed in a greenhouse during the summer season and then contaminated with the studied fungicides. Treatments consisted of different disinfection treatments, including a control without disinfection, solarization and biosolarization. For the solarization and biosolarization treatments, low-density polyethylene film was used as cover. Five pots per treatment were sampled periodically up to 90d after the beginning of each treatment and fungicide residues were analyzed by GC/MS. The results showed that both solarization and biosolarization enhanced fungicide dissipation rates with regard to the control treatment, an effect which was attributed to the increased soil temperature. Most of the fungicides studied showed similar behavior under solarization and biosolarization conditions. However, triadimenol was degraded to a greater extent in the biosolarization than in the solarization treatment, while fludioxonil behaved in the opposite way. The results confirm that both solarization and biosolarization contribute to pesticide dissipation and can therefore be considered alongside other soil disinfection techniques, as a bioremediation tool for pesticide-polluted soils.

Topics: Dioxoles; Environmental Restoration and Remediation; Fungicides, Industrial; Hot Temperature; Methacrylates; Phenylacetates; Photochemical Processes; Pyrimidines; Pyrroles; Soil; Soil Pollutants; Strobilurins; Triazoles

The increasing occurrence of QoI fungicide resistance in Plasmopara viticola (Berk. & MA Curtis) Berl. & DeToni populations is becoming a serious problem in the control of grapevine downy mildew. In Japan, the existence of QoI-fungicide-resistant P. viticola was reported in 2009.. The QoI fungicide resistance in P. viticola samples collected from vineyards in Japan in 2008 and 2009 was monitored. Resistant P. viticola were detected in the regions where QoI fungicides have been introduced in accordance with the pest management programme, whereas in Hokkaido vineyards, where QoI fungicides have not yet been introduced, QoI-fungicide-resistant P. viticola were not found.. Japan comprises thousands of islands and is physically isolated from other countries by the sea. Monitoring the emergence, incidence and distribution of QoI fungicide resistance in P. viticola populations in Japan is necessary to improve pest management strategies for downy mildew disease in Japanese vineyards.

Topics: Drug Resistance, Fungal; Fungicides, Industrial; Genotype; Japan; Methacrylates; Oomycetes; Pest Control; Phenylacetates; Plant Diseases; Polymorphism, Genetic; Pyrimidines; Strobilurins; Vitis

The molecular profile and the biological response of isolates of Pyricularia oryzae Cavara obtained from ctenanthe to two strobilurins (azoxystrobin, kresoxim-methyl) and the phenylpyridinamine fungicide fluazinam were characterized, and compared with isolates from rice plants. Five different isozymes (alpha-esterase, lactate, malate, isocitrate and sorbitol dehydrogenases) and five random decamer primers for RAPD-PCR were used to generate molecular markers. Using unweighted pair-group with arithmetic average analysis, ctenanthe isolates were found to form a separate group distinct from that of the rice isolates for both sets of markers. Amplified polymorphic sequences of mitochondrial cytochrome b that were digested with Fnu4HI or StyI revealed no differences among Pyricularia isolates at amino acid positions 143 or 129 which confer resistance to strobilurins in several fungi. In absence of the alternative respiration inhibitor salicylhydroxamic acid (SHAM) the three fungicides showed inferior and variable efficacy, with a trend toward the rice isolate being less sensitive. The addition of SHAM enhanced the effectiveness of all fungicides against isolates regardless of their origin. Appressorium formation was the most vulnerable target of action of the respiration inhibitors and azoxystrobin the most effective. This is the first report of a comparison between the molecular profiles and sensitivities to respiration inhibitors for Pyricularia oryzae isolates from a non-gramineous host and from rice.

Topics: Acrylates; Aminopyridines; Ascomycota; Drug Resistance, Fungal; Fungicides, Industrial; Isoenzymes; Marantaceae; Methacrylates; Oryza; Oxygen Consumption; Phenylacetates; Phylogeny; Pyrimidines; Salicylamides; Spores, Fungal; Strobilurins

Populations of the causal agent of wheat tan spot, Pyrenophora tritici-repentis, that are collected from fields frequently treated with reduced fungicide concentrations have reduced sensitivity to strobilurin fungicides and azole fungicides (C14-demethylase inhibitors). Energy-dependent efflux transporter activity can be induced under field conditions and after in vitro application of sublethal amounts of fungicides. Efflux transporters can mediate cross-resistance to a number of fungicides that belong to different chemical classes and have different modes of action. Resistant isolates can grow on substrata amended with fungicides and can infect plants treated with fungicides at levels above recommended field concentrations. We identified the hydroxyflavone derivative 2-(4-ethoxy-phenyl)-chromen-4-one as a potent inhibitor of energy-dependent fungicide efflux transporters in P. tritici-repentis. Application of this compound in combination with fungicides shifted fungicide-resistant P. tritici-repentis isolates back to normal sensitivity levels and prevented infection of wheat leaves. These results highlight the role of energy-dependent efflux transporters in fungicide resistance and could enable a novel disease management strategy based on the inhibition of fungicide efflux to be developed.

Topics: Acrylates; Ascomycota; ATP-Binding Cassette Transporters; Drug Resistance, Fungal; Epoxy Compounds; Flavones; Fungicides, Industrial; Gene Expression Regulation, Fungal; Methacrylates; Microbial Sensitivity Tests; Phenylacetates; Plant Diseases; Plant Leaves; Pyrimidines; Strobilurins; Triazoles; Triticum

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

In yeast the resistance to kresoxim-methyl and azoxystrobin, like the resistance to strobilurin A (mucidin) is under the control of both mitochondrial cob gene and the PDR network of nuclear genes involved in multidrug resistance. The mucidin-resistant mucl (G137R) and muc2 (L275S) mutants of Saccharomyces cerevisiae containing point mutations in mtDNA were found to be cross-resistant to kresoxim-methyl and azoxystrobin. Cross-resistance to all three strobilurin fungicides was also observed in yeast transformants containing gain-of-function mutations in the nuclear PDR3 gene. On the other hand, nuclear mutants containing disrupted chromosomal copies of the PDR1 and PDR3 genes or the PDR5 gene alone were hypersensitive to kresoxim-methyl, azoxystrobin and strobilurin A. The frequencies of spontaneous mutants selected for resistance either to kresoxim-methyl, azoxystrobin or strobilurin A were similar and resulted from mutations both in mitochondrial and nuclear genes. The results indicate that resistance to strobilurin fungicides, differing in chemical structure and specific activity, can be caused by the same molecular mechanism involving changes in the structure of apocytochrome b and/or increased efflux of strobilurins from fungal cells.

Topics: Acrylates; Antifungal Agents; ATP-Binding Cassette Transporters; Cell Nucleus; DNA-Binding Proteins; Drug Resistance, Multiple, Fungal; Fatty Acids, Unsaturated; Fungal Proteins; Fungicides, Industrial; Genes, Fungal; Methacrylates; Microbial Sensitivity Tests; Mitochondria; Mutation; Phenylacetates; Pyrimidines; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Strobilurins; Trans-Activators; Transcription Factors

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