Strobilurin Fungicides Research Articles

Effect of Moisture, Organic Matter, Microbial Population and Fortification Level on Dissipation of Pyraclostrobin in Soils

The dissipation of pyraclostrobin, a strobilurin fungicide, in soil was found to be influenced by soil moisture, organic matter content and microbial population. Among the different moisture regimes, dissipation was faster under submerged condition (T1/2 10 days) followed by field capacity (T1/2 28.7 days) and in dry soil (T1/2 41.8 days). Use of sludge at 5 % level to Inceptisol favoured a faster dissipation of pyraclostrobin, whereas a slower rate of dissipation was observed in partial organic matter removed soil as compared to normal soil. Slower rate of dissipation was also observed in sterile soil (T1/2 47 days) compared to normal soil. Pyraclostrobin dissipated faster in Vertisol (T1/2 21.8 days) than in Inceptisol (T1/2 28.7 days). No significant difference in the dissipation rate was observed at 1 and 10 μg g(-1) fortification levels.

Carbon nanotube-protein carriers enhance size-dependent self-adjuvant antibody response to haptens

Carbon nanotubes (CNTs) are nanomaterials with interesting emerging applications. Their properties make CNTs excellent candidates for use as new nanovehicles in drug delivery, immunization and diagnostics. In the current study, we assessed the immune-response-amplifying properties of CNTs to haptens by using azoxystrobin, the first developed strobilurin fungicide, as a model analyte. An azoxystrobin derivative bearing a carboxylated spacer arm (hapten AZc6) was covalently coupled to bovine serum albumin (BSA), and the resulting BSA–AZc6 conjugate was covalently linked to four functionalized CNTs of different shapes and sizes, varying in diameter and length. These four types of CNT-based constructs were obtained using efficient, fast, and easy functionalization procedures based on microwave-assisted chemistry. New Zealand rabbits and BALB/c mice were immunized with BSA–AZc6 alone and with the four CNT–BSA–AZc6 constructs, both with and without Freund's adjuvant. The IgG-type antibody responses were assessed in terms of the titer and affinity, paying special attention to the relationship between the immune response and the size and shape of the employed CNTs. Immunization with CNT–BSA–AZc6 resulted in enhanced titers and excellent affinities for azoxystrobin. More important, remarkable IgG responses were obtained even in the absence of an adjuvant, thus proving the self-adjuvanting capability of CNTs. Immunogens were able to produce strong anti-azoxystrobin immune responses in rabbits even when administered at a BSA–AZc6 conjugate dose as low as 0.05μg. The short and thick CNT–BSA–AZc6 construct produced the best antibody response under all tested conditions.

Efficacy loss of strobilurins used in protection against apple scab in Czech orchards

The apple scab caused by Venturia inaequalis is important apple disease in temperate zone. Economical losses caused by the pathogen are effectively reduced by intensive chemical protection and breeding. Occurrences of pathogen resistence to strobilurin fungicides were noted in some Czech orchards in the past. In 2007–2011, collection of 136 isolates V. inaequalis was tested with the aim to detect presence of single-point mutations of cytochrom b (cytb) gene causing efficacy loss of strobilurins. Consequently occurrence of beta-tubulin (beta-tub) gene mutation causing resistance to benomyl was studied with the aim to obtain comparative data about frequency of mutation in non-selective environment. Therefore simple and highly repeatable PCR based mutation detecting methods were optimised. Analysis brought knowledge about high similarity of cytb and beta-tub genes mutations occurrence. Whilst the frequency of the G143A substitution in cytb gene was 65% the E198A substitution of beta-tub gene occurred in 54% within pathogen population. This high frequency of resistant population necessitates revision of the way of strobilurins use in apple scab management.    

Assessing fungicide resistance in populations of Alternaria in Idaho potato fields

Early blight, caused by the fungus Alternaria solani and brown leaf spot, caused by Alternaria alternata, are important diseases of potato crops in Idaho. In recent years growers have reported a reduction in efficacy of fungicides traditionally used in the past decade to control early blight. In 2009, a collection of A. solani 39 isolates were screened for resistance to azoxystrobin, pyraclostrobin, boscalid and famoxadone. Fungicide sensitivity testing was done using spiral plate dilution gradients. Results showed that of 39 isolates screened, all were resistant to azoxystrobin and three were resistant to boscalid. None were resistant to pyraclostrobin or famoxadone. In summer 2010, more isolates were collected (9 A. alternata and 26 A. solani) and the survey was expanded to include more fungicides with four different modes of action that targeted succinate dehydrogenase (SDH), methionine biosynthesis, mitochondrial respiration and multi-site contact activity. New isolates of A. solani and A. alternata were also collected from two additional sites. The results showed that 57% of the isolates were resistant to boscalid as well as an average of 63% of the isolates being resistant to the strobilurin fungicides. Seven and 15% of isolates were resistant to penthiopyrad (an SDH inhibitor), and pyrimethanil (a methionine biosynthesis inhibitor), respectively. However, none of the isolates were resistant to fluopyram (an SDH inhibitor) or a mixture of fluopyram and pyrimethanil. Although there appears to be cross resistance developing in Alternaria spp. to some of the new SDH inhibitors like penthiopyrad, others such as fluopyram are still showing limited to no resistance development in Alternaria spp. in Idaho.

Ultrasound-assisted surfactant-enhanced emulsification microextraction with solidification of floating organic droplet followed by high performance liquid chromatography for the determination of strobilurin fungicides in fruit juice samples

A novel method, ultrasound-assisted surfactant-enhanced emulsification microextraction with solidification of floating organic droplet (UASEME-SFOD), has been developed for the extraction of four strobilurin fungicides (kresoxim-methyl, picoxystrobin, pyraclostrobin and trifloxystrobin) in fruit juices. In the UASEME-SFOD technique, Tween 80 was used as emulsifier, and 1-undecanol was used as extraction solvent without using any organic dispersive solvent. Several parameters that affect the extraction efficiency, such as the kind and volume of extraction solvent, the type and concentration of the surfactant, extraction time, extraction temperature and salt addition were investigated and optimized. Under the optimum extraction condition, the method yields a linear calibration curve in the concentration range from 5 to 10,000 ng mL(-1) for the targeted analytes with the correlation coefficient ranging from 0.9991 to 0.9998. The enrichment factors were in the range between 95 and 135, and the limits of detection of the method were 2-4 ng mL(-1). The fruit juice samples were successfully analyzed using the proposed method, and the relative recoveries at fortified levels of 50 and 100 ng mL(-1) were in the range of 82.6-97.5%.

Determination of Strobilurin Fungicide Residues in Fruits and Vegetables by Micellar Electrokinetic Capillary Chromatography with Sweeping

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.

Occurrence and persistence of fungicides in bed sediments and suspended solids from three targeted use areas in the United States

To document the environmental occurrence and persistence of fungicides, a robust and sensitive analytical method was used to measure 34 fungicides and an additional 57 current-use pesticides in bed sediments and suspended solids collected from areas of intense fungicide use within three geographic areas across the United States. Sampling sites were selected near or within agricultural research farms using prophylactic fungicides at rates and types typical of their geographic location. At least two fungicides were detected in 55% of the bed and 83% of the suspended solid samples and were detected in conjunction with herbicides and insecticides. Six fungicides were detected in all samples including pyraclostrobin (75%), boscalid (53%), chlorothalonil (41%) and zoxamide (22%). Pyraclostrobin, a strobilurin fungicide, used frequently in the United States on a variety of crops, was detected more frequently than p,p′-DDE, the primary degradate of p,p′-DDT, which is typically one of the most frequently occurring pesticides in sediments collected within highly agricultural areas. Maximum fungicide concentrations in bed sediments and suspended solids were 198 and 56.7μg/kg dry weight, respectively. There is limited information on the occurrence, fate, and persistence of many fungicides in sediment and the environmental impacts are largely unknown. The results of this study indicate the importance of documenting the persistence of fungicides in the environment and the need for a better understanding of off-site transport mechanisms, particularly in areas where crops are grown that require frequent treatments to prevent fungal diseases.

Simultaneous determination of three strobilurin fungicide residues in fruits, vegetables and soil by a modified quick, easy, cheap, effective, rugged (QuEChERS) method coupled with gas chromatography-tandem mass spectrometry

A gas chromatography-tandem mass spectrometry (GC-MS/MS) method followed by a modified quick, easy, cheap, effective, rugged (QuEChERS) method for the analysis of three strobilurin fungicide residues (picoxystrobin, kresoxim-methyl and trifloxystrobin) in fruits, vegetables and soil was developed and validated. The three fungicides were extracted with ethyl acetate and cyclohexane (v/v = 1/1), and subsequently cleaned up using a combination of primary secondary amine and graphitized carbon black prior to GC-MS/MS analysis. The three fungicides were qualitatively determined using the multiple reaction monitoring mode with two ion transitions measured for each fungicide and evaluation by collision tests, and were determined quantitatively using the external standard method. Recovery studies were performed at three fortification levels of 0.005, 0.05, and 0.5 mg kg−1 in blank cucumber, tomato, apple, grape and soil samples. The overall average recoveries ranged from 87% to 112%, with intra-day relative standard deviations (RSDs) of 0.9% to 10.5% and inter-day RSD of 0.7% to 12.2%, respectively. The calculated limits of detection and quantification were typically below 0.006 and 0.019 μg kg−1 for GC-MS/MS. The results of the method validation confirm that this proposed method is easy and reliable for the determination of residues of strobilurin fungicides in fruits, vegetables and soil.

Determination of seven strobilurin fungicide residues in Chinese herbs by liquid chromatography-tandem mass spectrometry coupled with solid phase extraction

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).

Effect of eradicative activity of strobilurin fungicides and difenconazole on mycelium and sporulation of Venturia inaequalis

The eradicative effect of two strobilurin fungicides (kresoxim-methyl and trifloxystrobin) and difenoconazole on activity of apple scab spots was studied in greenhouse and field conditions. The reaction of mycelium and sporulation of <i>Venturia inaequalis</i> after fungicides applications were observed in light and scanning electron microscope. Mixture of strobilurin fungicide and difenoconazole applied twice, in 5 days interval, gave the best eradicative effect. The inhibition of sporulation was long lasting and it was observed even 3 and 5 weeks after treatments. The conidiophores, conidia and mycelium were damaged. Also difenoconazole showed good eradicative activity, in field condition up to 5 weeks after application. Trifloxystrobin significantly reduced the number of active scab lesions and the effect was visible even 3 weeks after treatment. Kresoxim-methyl in both tested doses (0,2 and 0,3 kg/ha) was slightly less effective.

Quantifying the Effect of Pyraclostrobin on Grainfill Period and Kernel Dry Matter Accumulation in Maize

Strobilurin fungicides are effective against a wide range of foliar fungal diseases on several crops and may offer additional physiological benefits, including plants staying green longer than normal (the “stay-green effect”). It has been hypothesized that the stay-green effect may extend the grain fill period leading to increased grain yield due to a longer period of dry matter accumulation. We investigated the effect of pyraclostrobin fungicide applied at tasseling on foliar disease suppression, stalk rot severity, the stay-green effect of leaves in the upper canopy, dry matter accumulation, time at physiological maturity, grain yield, and moisture at harvest in maize from 2008 through 2010 in Iowa at six location years. Foliar disease severity was <5% in all location years. Pyraclostrobin-treated plots had a significantly higher area under green leaves incidence curve compared to non-treated maize. Although grain yield and grain moisture at harvest were not different (P > 0.1) between pyraclostrobin-treated and non-treated maize in all location years, treated plots tended to have higher yield and grain moisture. Time at physiological maturity did not differ between pyraclostrobin-treated and non-treated plots (P > 0.1). Although we demonstrated an application of pyraclostrobin to maize delayed senescence of the leaves thus contributing to the stay-green effect, our data did not show grain-fill period extension. Accepted for publication 21 August 2013. Published 24 October 2013.

Metabolism of Strobilurins by Wheat Cell Suspension Cultures

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.

Interaction between seed treatments, surfactants and foliar fungicides on controlling dry bean anthracnose (Colletotrichum lindemuthianum)

Anthracnose, caused by fungal pathogen Colletotrichum lindemuthianum (Sacc. & Magnus) Briosi Cav. is one of the main production constraints of the dry bean (Phaseolus vulgaris L.) industry in Ontario. A field study was carried out in 2007 and 2008 to investigate the effect of two seed treatments (DCT (diazinon + captan + thiophanate-methyl) and MFA (metalaxyl-M + fludioxonil + azoxystrobin)) and two foliar fungicides (pyraclostrobin and azoxystrobin) applied with and without a surfactant under low and high disease pressure conditions at Exeter ON. Eighteen treatment combinations were tested in a randomized complete block design with four replicates. The treatment effects were examined by measuring disease development on leaf and pod tissue, pod destruction index, pick (discolored and misshaped seed), yield and return on investment (ROI). The seed treatment MFA performed similarly to DCT, and should be considered a suitable replacement to DCT for dry bean growers. However, utilizing a strobilurin fungicide in both seed and foliar treatments raises concern, as this practice increases the risk of disease resistance. The addition of a surfactant to azoxystrobin increased seed yield and ROI under high disease pressure, but had no effect when added to pyraclostrobin. Pyraclostrobin outperformed azoxystrobin for some disease indices as well as for yield under high disease pressure and for ROI under low and high disease pressure conditions.

ChemInform Abstract: Synthesis and Antifungal Activities of New Type β‐Methoxyacrylate‐Based Strobilurin Analogues.

Strobilurins have become one of the most important classes of agricultural fungicides. To discover new strobilurin derivatives with high activity against resistant pathogens, a series of novel beta-methoxyacrylate analogues were designed and synthesized by integrating substituted pyrimidine with a strobilurin pharmacophore. The compounds were confirmed and characterized by infrared, 1H nuclear magnetic resonance, elemental analysis and mass spectroscopy. The bioassays indicated that most of the compounds 1 exhibited potent antifungal activity against Colletotrichum orbiculare, Botrytis cinerea Pers and Phytophthora capsici Leonian at the concentration of 50 mu g/mL. Exhilaratingly, compound 1a (R=methyl) showed better antifungal activity against all the tested fungi than the commercial strobilurin fungicide azoxystrobin.

Design, Synthesis, and Antifungal Activities of New β‐Methoxyacrylate Analogues

AbstractStrobilurins have become one of the most important classes of agricultural fungicides. To search for new strobilurin derivatives with high activity against resistant pathogens, a series of new β‐methoxyacrylate analogues containing substituted pyrimidine in the side chain with strobilurin pharmacophore were synthesized and their biological activities were tested. The compounds were confirmed and characterized by 1H‐NMR, elemental analysis and mass spectroscopy. The bioassays indicated that most of the compounds 1 exhibited potent antifungal activities against Colletotrichum orbiculare, Botrytis cinerea Pers and Phytophthora capsici Leonian at a concentration of 50 μg mL−1. Notably, compound 1b (R = 2,5‐dimethylphenyl) showed better antifungal activity against all the tested fungi than the commercial strobilurin fungicide azoxystrobin.

Direct surface plasmon resonance immunosensing of pyraclostrobin residues in untreated fruit juices

A surface plasmon resonance (SPR) immunoassay for on-line detection of the strobilurin fungicide pyraclostrobin in untreated fruit juices is presented. The analysis of pyraclostrobin residues is accomplished in apple, grape, and cranberry samples by monitoring the recognition events occurring separately in a two-channel home-made SPR biosensor. Covalent coupling of the analyte derivative results in a reversible method, enabling more than 80 measurements on the same sensor surface. Optimization of the immunoassay conditions provides limits of detection as low as 0.16 μg L(-1). The selectivity and reproducibility of the analysis is ensured by studying both non-specific interactions with unrelated compounds and inter-assay coefficients of variation. Excellent recovery ranging from 98 to 103% was achieved by a simple 1:5 dilution of fruit juice with assay buffer before the analysis. The lack of previous cleaning and homogenization procedures reduces the analysis time of a single food sample to only 25 min, including the regeneration cycle.

Design, Synthesis and Antifungal Activities of Novel Strobilurin Derivatives Containing Pyrimidine Moieties

Strobilurins are one of the most important classes of agricultural fungicides. To discover new strobilurin derivatives with high activity against resistant pathogens, a series of novel beta-methoxyacrylate analogues were designed and synthesized by integrating substituted pyrimidine with a strobilurin pharmacophore. The compounds were confirmed and characterized by infrared, H-1 nuclear magnetic resonance, elemental analysis and mass spectroscopy. The bioassays indicated that most of the compounds (1a-1h) exhibited potent antifungal activities against Colletotrichum orbiculare, Botrytis cinerea Pers and Phytophthora capsici Leonian at the concentration of 50 mu g/mL. Exhilaratingly, compound 1d (R=3-trifluoromethylphenyl) showed better antifungal activity against all the tested fungi than the commercial strobilurin fungicide azoxystrobin.

Viabilidade técnica e econômica da aplicação de estrobilurinas em milho

Technical and economic feasibility of strobilurin fungicides application on maize In recent years, great emphasis has been given to the use of fungicides for managing foliar diseases in corn in Brazil. This study aimed to conduct an analysis of technical and economic feasibility of application of fungicides on maize in different growing regions. Trials were conducted to evaluate the effect of fungicides on the yield of different maize cultivars at three locations: Sete Lagoas (state of Minas Gerais), Londrina (state of Parana) and Rio Verde (state of Goias). The effect of one and two fungicide applications on each cultivar was evaluated. For each trial, the difference in mean yield and disease severities between treated and untreated plots for each cultivars was compared. For all trials, yield response of cultivars treated with fungicides was highly inconsistent under low disease severity. A higher frequency of positive yields and economic benefit occurred when fungicide applications were made under conditions of high disease pressure. More studies are needed to better clarify the effect of strobilurin fungicides on the physiology and production of corn plants.

Synthesis and Antifungal Activities of New Type β‐Methoxyacrylate‐Based Strobilurin Analogues

AbstractStrobilurins have become one of the most important classes of agricultural fungicides. To discover new strobilurin derivatives with high activity against resistant pathogens, a series of novel β‐methoxyacrylate analogues were designed and synthesized by integrating substituted pyrimidine with a strobilurin pharmacophore. The compounds were confirmed and characterized by infrared, 1H nuclear magnetic resonance, elemental analysis and mass spectroscopy. The bioassays indicated that most of the compounds 1 exhibited potent antifungal activity against Colletotrichum orbiculare, Botrytis cinerea Pers and Phytophthora capsici Leonian at the concentration of 50 μg/mL. Exhilaratingly, compound 1a (R=methyl) showed better antifungal activity against all the tested fungi than the commercial strobilurin fungicide azoxystrobin.

Acute toxicity of three strobilurin fungicide formulations and their active ingredients to tadpoles

Fungicide applications in the United States have increased tenfold in the last 5 years. Formulations and active ingredients (AIs) have been demonstrated to cause acute mortality to amphibian life stages. However, there has been little to no discrimination between the toxicity of fungicide formulations and their AIs. Therefore, we compared the acute toxicity of the active ingredients and formulations of the fungicides Headline®, Stratego®, and Quilt® using Bufo cognatus tadpoles exposed to four concentrations and a control. All fungicides, including AIs and formulations, demonstrated toxicity to tadpoles, with Headline® and Stratego® causing 100 % mortality at the highest concentrations. Exposure to Quilt® formulation and its AIs resulted in 50-60 % tadpole mortality. Overall, toxicity was comparable between AIs and formulations for all fungicides and concentrations, with the exception of Headline® at 5 μg/L, where formulation exposure resulted in 79 % mortality versus no mortality from exposure to the AI. Results suggest the AIs are responsible for most mortality for Quilt® and Stratego®. Results for Headline® however suggest that although the AI is toxic to tadpoles at environmentally relevant concentrations, adjuvant(s) in the Headline® formulation also contribute to mortality, making it the most toxic of the fungicides studied.