Two types of amino acid substitutions in the succinate dehydrogenase complex subunit confer resistance to benzovindiflupyr in Colletotrichum sublineola.

BACKGROUND: Colletotrichum sublineola is the pathogenic fungus that causes sorghum anthracnose, which seriously threatens sorghum yield. Benzovindiflupyr is a succinate dehydrogenase inhibitor with good control effects on various crop diseases. However, the control of sorghum anthracnose by benzovindiflupyr and the risk of resistance to benzovindiflupyr in this pathogen are not well studied. Therefore, this study aimed to evaluate the benzovindiflupyr resistance and underlying mechanisms in C. sublineola. RESULTS: Analysis of the sensitivity of 126 C. sublineola strains to benzovindiflupyr revealed that the average EC50 of the fungicide was 0.0503 ± 0.0189 µg mL-1, with a unimodal normal distribution curve. The survival fitness of 10 benzovindiflupyr-resistant strains decreased to varying degrees compared with that of the wild-type parental strains. Additionally, a significant positive cross-resistance was observed between benzovindiflupyr and carboxin. Sequencing analyses identified two mutation sites, CsSdhBH249Y and CsSdhCG81V, in the resistant strains. Further molecular docking and site-directed mutagenesis experiments confirmed that the CsSdhBH249Y and CsSdhCG81V substitutions conferred resistance to benzovindiflupyr in C. sublineola. CONCLUSION: Colletotrichum sublineola is sensitive to benzovindiflupyr and shows a moderate resistance risk to benzovindiflupyr. Two specific point substitutions, CsSdhBH249Y and CsSdhCG81V, are responsible for the resistance of C. sublineola to benzovindiflupyr. These findings offer a theoretical foundation for strategic application of the fungicide in controlling sorghum anthracnose, and for potentially delaying the emergence and progression of resistance. © 2024 Society of Chemical Industry.

Amino acid mutation of succinate dehydrogenase complex induced resistance to benzovindiflupyr in Magnaporthe oryzae.

Magnaporthe oryzae is a rice blast pathogen that seriously threatens rice yield. Benzovindiflupyr is a succinate dehydrogenase inhibitor (SDHI) fungicide that effectively controls many crop diseases. Benzovindiflupyr has a strong inhibitory effect on M. oryzae; however, control of rice blast by benzovindiflupyr and risk of resistance to benzovindiflupyr are not well studied in this pathogen. In this study, six benzovindiflupyr-resistant strains were obtained by domestication induced in the laboratory. The MoSdhBH245D mutation was the cause of M. oryzae resistance to benzovindiflupyr, which was verified through succinate dehydrogenase (SDH) activity assays, molecular docking, and site-specific mutations. Survival fitness analysis showed no significant difference between the benzovindiflupyr-resistant and parent strains. Positive cross-resistance to benzovindiflupyr and other SDHIs and negative cross-resistance to azoxystrobin were observed. Therefore, the risk of benzovindiflupyr resistance in M. oryzae might be medium to high. It should be combined with other classes of fungicides (tebuconazole and azoxystrobin) to slow the development of resistance.

Adsorption-desorption and leaching behavior of benzovindiflupyr in different soil types.

Benzovindiflupyr is a succinate dehydrogenase inhibitor fungicide that targets mitochondrial function for disease control. In this study, we investigated the adsorption-desorption and leaching behavior of benzovindiflupyr in eight soil types using the batch equilibrium method and the soil column leaching method. A Freundlich model (r2 > 0.9959) was used to better characterize the adsorption-desorption process in eight soil types, with adsorption coefficients (KF-ads) ranging from 2.303 to 17.886. KF-ads was significantly and positively correlated (p < 0.05) with the organic carbon content. High temperatures and increased initial pH of aqueous solutions led to a decrease in benzovindiflupyr adsorption in the soil. The adsorption was also influenced by factors such as ionic strength, humic acid, surfactant type, microplastic type, and particle size and concentration. Moreover, benzovindiflupyr exhibited low leachability in all four soils selected, but different leaching solutions affected the risk of benzovindiflupyr migration to groundwater. Overall, this study provides insights into the adsorption characteristics of benzovindiflupyr in different soils and provides key information for environmental risk assessment.

Study on the enantioselective behaviors, activity, toxicity and mechanism of novel SDHI fungicide benzovindiflupyr to reduce the environmental risks.

Benzovindiflupyr (BEN) has emerged as one of the fastest-growing SDHI fungicides in recent years, but it is considered "very highly toxic" to aquatic fish, invertebrates and crustaceans (EC50 or LC50, 0.0035-0.056 mg/L, acute toxicity). The comprehensive study on bioactivity, toxicity, and degradation behaviors of BEN at the enantiomeric level would facilitate the development of a high-efficiency and low-risk application method. The bioactivities of 1S, 4R-(-)-BEN against five target pathogens (Alternaria alternata, Phoma multirostrata, Selerotium rolfsii, Magnaporthe oryzae, and Rhizoctonia solani) (EC50, 0.00562-0.329 mg/L, high-efficiency) were 6.7-1029 times higher than 1R, 4S-(+)-BEN, demonstrating significant enantioselectivity. For Danio rerio, 1S, 4R-(-)-BEN (LC50, 0.0360 mg/L, "very highly toxic") exhibited higher toxicity than 1 R, 4S-(+)-BEN, but the toxic interaction was concentration addition (TUrac, 0.94), indicating an enhanced toxicity in the presence of 1R, 4S-(+)-BEN. Molecular docking was employed to offer insights at the molecular level and elucidate the factors influencing enantioselectivity. The stronger binding affinity of 1S, 4R-(-)-BEN with SDH was in line with the quantitative experimental findings. The degradation of two BEN enantiomers in four different fruits followed the first-order degradation kinetics equation, and displayed enantioselectivity. The preferential degradation of 1R, 4S-(+)-BEN was found in pears and grapes, while varying enantioselectivity was found at different stages in tomatoes and watermelons. The residual concentrations of BEN in grapes were higher than the EU's MRL, which in the other three fruits were below the MRLs during the sampling. In conclusion, 1S, 4R-(-)-BEN proved to be the more effective monomer. Utilizing the pure monomer could not only reduce the dosage of racemate by about 44-59 %, but also mitigate the risk of introducing inefficient monomer into the environment (especially for fish).

Resistance risk assessment for benzovindiflupyr in Sclerotium rolfsii and transmission of resistance genes among population.

BACKGROUND: Sclerotium rolfsii is a destructive soil-borne fungal pathogen which is distributed worldwide. In previous study, the succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr has been identified for its great antifungal activity against Sclerotium rolfsii. This study is aimed to investigate the resistance risk and mechanism of benzovindiflupyr in Sclerotium rolfsii. RESULTS: Eight stable benzovindiflupyr-resistant isolates were generated by fungicide adaptation. Although the obtained eight resistant isolates have a stronger pathogenicity than the parental sensitive isolate, they have a fitness penalty in the mycelial growth and sclerotia formation compared to the parental isolate. A positive cross-resistance existed in the resistant isolates between benzovindiflupyr and thifluzamide, carboxin, boscalid and isopyrazam. Three-point mutations, including SdhBN180D, SdhCQ68E and SdhDH103Y, were identified in the benzovindiflupyr-resistant isolates. However, molecular docking analysis indicated that only SdhDH103Y could influence the sensitivity of Sclerotium rolfsii to benzovindiflupyr. After mycelial co-incubation of resistant isolates and the sensitive isolate, resistance genes may be transmitted to the sensitive isolate. The in vivo efficacy of benzovindiflupyr and thifluzamide against benzovindiflupyr-resistant isolates was a little lower than that against the sensitive isolate but with no significant difference. CONCLUSION: The results suggested a low to medium resistance risk of Sclerotium rolfsii to benzovindiflupyr. However, once resistance occurs, it is possible to spread in the population of Sclerotium rolfsii. This study is helpful to understanding the risk and mechanism of resistance to benzovindiflupyr in multinucleate pathogens such as Sclerotium rolfsii. © 2024 Society of Chemical Industry.

Activity of the Succinate Dehydrogenase Inhibitor Fungicide Benzovindiflupyr Against Clarireedia spp.

Dollar spot (DS), caused by Clarireedia spp. (formerly Sclerotinia homoeocarpa), is one of the most important diseases of turfgrasses worldwide. Benzovindiflupyr, a pyrazole carboxamide fungicide belonging to succinate dehydrogenase inhibitors, was recently registered for DS control. In this study, baseline sensitivity, toxicity, and control efficacy of benzovindiflupyr against Clarireedia spp. were evaluated. The frequency of sensitivities had a unimodal distribution (Kolmogorov-Smirnov, P > 0.10). The mean EC50 value was 1.109 ± 0.555 µg/ml, with individual values ranging from 0.160 to 2.548 µg/ml. Benzovindiflupyr increased the number of hyphal offshoots and cell membrane permeability and inhibited oxalic acid production. Positive cross-resistance was observed between benzovindiflupyr and boscalid, but not between benzovindiflupyr and thiophanate-methyl, propiconazole, or iprodione. Benzovindiflupyr showed high protective and curative control efficacies in vivo and in field applications. Both protective and curative control efficacies of benzovindiflupyr were significantly better than propiconazole, and equivalent to boscalid, over 2 years of field research. The results have important implications for managing DS and fungicide resistance problems in Clarireedia spp.

Absolute Configuration, Enantioselective Bioactivity, and Mechanism Study of the Novel Chiral Fungicide Benzovindiflupyr.

Benzovindiflupyr has gained increasing attention as a new chiral succinate dehydrogenase inhibitor fungicide; however, its determination, bioactivity, and mechanism at the enantiomeric level are very limited. In the present study, optical rotation determination and X-ray single-crystal diffraction results identified that the absolute configurations were (+)-(1R,4S)-benzovindiflupyr and (-)-(1S,4R)-benzovindiflupyr. A quantitative determination method for enantiomers was established using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) for pesticide detection. The stereoselective bioactivity assay indicated that (-)-(1S,4R)-benzovindiflupyr exhibited greater potency than (+)-(1R,4S)-benzovindiflupyr against seven phytopathogenic fungi. Molecular docking analysis showed that (-)-(1S,4R)-benzovindiflupyr possessed a stronger binding affinity to succinate dehydrogenase than (+)-(1R,4S)-benzovindiflupyr. The binding modes between enantiomers and the mutant with H272(B) predicted that the phytopathogenic fungi with H272(B) of succinate dehydrogenase mutation would not be resistant to benzovindiflupyr enantiomers. This study provides a basis for residue evaluation, risk assessment, and the safe application of benzovindiflupyr at the enantiomer level.

Bioactivity and Control Efficacy of Benzovindiflupyr Against Athelia rolfsii in China.

Athelia rolfsii is a devastating soilborne pathogen that causes stem rot of peanut and severely restricts peanut production. The new generation of succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr has been registered in the United States and Brazil for managing multiple plant diseases. However, it is not registered in China to control peanut stem rot. In this study, 246 isolates from major peanut production areas in Shandong, Henan, and Hebei Provinces of China were used to determine the baseline sensitivity of A. rolfsii to benzovindiflupyr. The frequency of EC50 values of benzovindiflupyr was unimodally distributed with an average EC50 of 0.12 ± 0.05 mg/liter and a range of 0.01 to 0.57 mg/liter. Benzovindiflupyr can also strongly inhibit the germination of sclerotia, with an average EC50 of 2.38 ± 1.04 mg/liter (n = 23). In addition, benzovindiflupyr exhibited great in vivo efficacy against A. rolfsii; the protective or curative efficacy (89.87%, 20.39%) of benzovindiflupyr at a concentration of 50 mg/liter was equivalent to that of the control fungicide thifluzamide at 100 mg/liter (86.39%, 16.21%). At the same concentration (e.g., 100 mg/liter), the protective efficacy (93.99%) of benzovindiflupyr was more than twice as high as the curative efficacy (45.07%). A positive correlation existed between benzovindiflupyr and isopyrazam or mefentrifluconazole, which possibly resulted from similar chemical structures or damage to the cell membrane. Our findings provide valuable information for the application of benzovindiflupyr, and the established baseline sensitivity could facilitate the monitoring and assessment of benzovindiflupyr resistance risk.

Identification and Fungicide Sensitivity of Colletotrichum spp. from Apple Flowers and Fruitlets in Brazil.

Glomerella leaf spot (GLS) and bitter rot (BR), caused by Colletotrichum spp., are major diseases on apple in southern Brazil. Among integrated pest management tools for disease management in commercial orchards, fungicides remain an important component. This study aimed to identify Colletotrichum spp. from cultivar Eva in Paraná state orchards; evaluate their in vitro sensitivity to cyprodinil, tebuconazole, iprodione, and fluazinam; and determine the baseline in vitro sensitivity of these isolates to benzovindiflupyr and natamycin. Most isolates belonged to Colletotrichum melonis and C. nymphaeae of the C. acutatum species complex. The two species varied in sensitivity to fluazinam and tebuconazole, but no variability was found for any other fungicide. The lowest 50% effective concentration (EC50) values of Colletotrichum spp. were observed for cyprodinil (mean EC50 < 0.02) and benzovindiflupyr (mean EC50 < 0.05); EC50 values were intermediate for fluazinam (mean EC50 < 0.33) and tebuconazole (mean EC50 < 0.14), and they were highest for natamycin (mean EC50 < 5.56) and iprodione (mean EC50 > 12). Cyprodinil and fluazinam are registered for use in Brazil for apple disease management but not specifically for GLS and BR. Tebuconazole is one of the few products registered for Colletotrichum spp. control in apples. In conclusion, flowers and fruitlets can serve as sources of inoculum for GLS and BR disease; C. acutatum was the predominant species complex in these tissues; cyprodinil and fluazinam applications may suppress GLS and BR; and benzovindiflupyr and natamycin warrant further investigation for GLS and BR disease control of apple due to comparably high in vitro sensitivity.

Structural Elucidation of Agrochemicals and Related Derivatives Using Infrared Ion Spectroscopy.

Agrochemicals frequently undergo various chemical and metabolic transformation reactions in the environment that often result in a wide range of derivates that must be comprehensively characterized to understand their toxicity profiles and their persistence and outcome in the environment. In the development phase, this typically involves a major effort in qualitatively identifying the correct chemical isomer(s) of these derivatives from the many isomers that could potentially be formed. Liquid chromatography-mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy are often used in attempts to characterize such environment transformation products. However, challenges in confidently correlating chemical structures to detected compounds in mass spectrometry data and sensitivity/selectivity limitations of NMR frequently lead to bottlenecks in identification. In this study, we use an alternative approach, infrared ion spectroscopy, to demonstrate the identification of hydroxylated derivatives of two plant protection compounds (azoxystrobin and benzovindiflupyr) contained at low levels in tomato and spinach matrices. Infrared ion spectroscopy is an orthogonal tandem mass spectrometry technique that combines the sensitivity and selectivity of mass spectrometry with structural information obtained by infrared spectroscopy. Furthermore, IR spectra can be computationally predicted for candidate molecular structures, enabling the tentative identification of agrochemical derivatives and other unknowns in the environment without using physical reference standards.

Sensitivity to fungicides in isolates of Colletotrichum gloeosporioides and C. acutatum species complexes and efficacy against anthracnose diseases.

Colletotrichum species cause diseases on many plants and are among the 'top 10' fungal plant pathogens. Species of the C. gloeosporioides and C. acutatum complexes are particularly important because they infect temperate fruit crops, but their control relies largely on chemical fungicides. In this study, differences in intrinsic fungicide sensitivity were determined in vitro using isolates of the C. gloeosporioides sp. complex (C. fructicola, C. siamense, and C. tropicale) and the C. acutatum sp. complex (C. fioriniae and C. nymphaeae), which had never been exposed to fungicides. Mycelial growth of all isolates was sensitive to the QoI azoxystrobin, the SDHI benzovindiflupyr, and the new DMI fungicide mefentrifluconazole. The isolates of C. nymphaeae were highly sensitive to the phenylpyrrole fungicide fludioxonil. The isolates of C. gloeosporioides sp. complex were sensitive to the bis-guanidine fungicide iminoctadine-albesilate, whereas those of C. acutatum sp. complex were inherently insensitive. These results are valuable when sensitivity of field populations is monitored in resistance management. Although SDHI fungicides are largely not effective against diseases caused by Colletotrichum species, benzovindiflupyr controlled anthracnose disease of various crops such as kidney bean, garland chrysanthemum, and strawberry, caused by C. lindemuthianum, C. chrysanthemi, and C. siamense, respectively, demonstrating this fungicide to be unique among SDHIs and having a broad control spectrum against anthracnose. To help understanding the reason for differential activity of benzovindiflupyr and boscalid, sdhB gene sequences were analyzed but those of C. lindemuthianum, C. chrysanthemi, and C. scovillei revealed no known mutations reported to be responsible for SDHI resistance in other fungi, indicating that other mechanism(s) than target-site modification may be involved in differential sensitivity to benzovindiflupyr and boscalid, found in Colletotrichum species.

Fungicide Sensitivity of Colletotrichum Species Causing Bitter Rot of Apple in the Mid-Atlantic U.S.A.

Apple growers in the Mid-Atlantic region of the U.S.A. have reported increased losses to bitter rot of apple. We tested the hypothesis that this increase is because the Colletotrichum population has developed resistance to commonly used single-mode-of-action (single-MoA) fungicides. We screened 220 Colletotrichum isolates obtained from 38 apple orchards in the Mid-Atlantic region for resistance to 11 fungicides in Fungicide Resistance Action Committee (FRAC) groups 1, 7, 9, 11, 12, and 29. Eleven (5%) of these isolates were resistant to FRAC group 1 with confirmed ß-tubulin E198A mutations, and two (<1%) were also resistant to FRAC group 11 with confirmed cytochrome-b G143A mutations. Such low frequencies of resistant isolates indicate that fungicide resistance is unlikely to be the cause of any regional increase in bitter rot. A subsample of isolates was subsequently tested in vitro for sensitivity to every single-MoA fungicide registered for apple in the Mid-Atlantic U.S.A. (22 fungicides; FRAC groups 1, 3, 7, 9, 11, 12, and 29), and 13 fungicides were tested in field trials. These fungicides varied widely in efficacy both within and between FRAC groups. Comparisons of results from our in vitro tests with results from our field trials and other field trials conducted across the eastern U.S.A. suggested that EC25 values (concentrations that reduce growth by 25%) are better predictors of fungicide efficacy in normal field conditions than EC50 values. We present these results as a guideline for choosing single-MoA fungicides for bitter rot control in the Mid-Atlantic U.S.A.

Comparative Transcriptome Analyses Reveal Conserved and Distinct Mechanisms of the SDHI Fungicide Benzovindiflupyr Inhibiting Colletotrichum.

Colletotrichum leaf disease (CLD) is an annual production concern for commercial growers worldwide. The succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr shows higher bioactivity against CLD than other SDHIs. However, the mechanism underlying such difference remains unclear. In this study, benzovindiflupyr exhibits good inhibitory activity against Colletotrichum siamense and C. nymphaeae in vitro and in vivo. To reveal its mechanism for inhibiting Colletotrichum, we compared transcriptomes of C. siamense and C. nymphaeae under treatment with benzovindiflupyr and boscalid. Benzovindiflupyr exhibited higher inhibitory activity against SDH enzyme than boscalid, resulting in a greater reduction in the ATP content of Colletotrichum isolates. Most of the metabolic pathways induced in these fungicide-treated isolates were similar, indicating that benzovindiflupyr exhibited a conserved mechanism of SDHIs inhibiting Colletotrichum. At the same level of suppressive SDH activity, benzovindiflupyr activated more than three times greater gene numbers of Colletotrichum than boscalid, suggesting that benzovindiflupyr could activate distinct mechanisms against Colletotrichum. Membrane-related gene ontology terms, mainly including intrinsic components of membrane, were highly abundant for the benzovindiflupyr-treated isolates rather than boscalid-treated isolates. Only benzovindiflupyr increased the relative conductivities of hyphae, indicating that it could damage the cell membrane and increase mycelial electrolyte leakage. Thus, we proposed that the high bioactivity of benzovindiflupyr against Colletotrichum occurred by inhibiting SDH activity and damaging the cell membrane at the same time. The research improves our understanding the mode of action of SDHI fungicides against Colletotrichum.

Modification of the existing maximum residue levels for benzovindiflupyr in fresh herbs and edible flowers.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta Agro GmbH submitted a request to the competent national authority in Germany to modify the existing maximum residue levels (MRLs) for the active substance benzovindiflupyr in fresh herbs and edible flowers. The data submitted in support of the request were found to be sufficient to derive MRL proposals for the crops under assessment. Adequate analytical methods for enforcement are available to control the residues of benzovindiflupyr on the commodities under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the short-term and long-term intake of residues resulting from the use of benzovindiflupyr according to the reported agricultural practices is unlikely to present a risk to consumer health.

Modification of the existing maximum residue levels for benzovindiflupyr in leeks and spring onions/green onions/Welsh onions.

In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta Crop Protection AG submitted a request to the competent national authority in France to modify the existing maximum residue levels (MRLs) for the active substance benzovindiflupyr in leeks and spring onions, green onions and Welsh onions. The data submitted in support of the request were found to be sufficient to derive MRL proposals for the crops under assessment. Adequate analytical methods for enforcement are available to control the residues of benzovindiflupyr on the commodities under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the short-term and long-term intake of residues resulting from the use of benzovindiflupyr according to the reported agricultural practices is unlikely to present a risk to consumer health.

Applying a Hybrid Modeling Approach to Evaluate Potential Pesticide Effects and Mitigation Effectiveness for an Endangered Fish in Simulated Oxbow Habitats.

The occurrence of some species listed under the United States' Endangered Species Act in agricultural landscapes suggests that their habitats could potentially be exposed to pesticides. However, the potential effects from such exposures on populations are difficult to estimate. Mechanistic models can provide an avenue to estimating the potential impacts on populations, considering realistic assumptions about the ecology of the species, the ecosystem it is part of, and the potential exposures within the habitat. In the present study, we applied a hybrid model of the Topeka shiner (Notropis topeka), a small endangered cyprinid fish endemic to the US Midwest, to assess the potential population-level effects of realistic exposures to a fungicide (benzovindiflupyr). The Topeka shiner populations were simulated in the context of the food web found in oxbow habitats that are the focus of ongoing habitat restoration efforts for the species. We applied realistic, time-variable exposure scenarios and represented lethal and sublethal effects to individual Topeka shiners using toxicokinetic-toxicodynamic models. With fish in general showing the highest sensitivity to the compound, direct effects on simulated Topeka shiner populations governed the population-level effects. We characterized the population-level effects of different exposure scenarios with exposure multiplication factors (EMFs) applied. The introduction of a vegetative filter strip (VFS; 15 ft; 4.6 m) between the treated area and the oxbow habitat was shown to be effective as mitigation because EMFs were 2 to 3 times higher than for the exposure scenario without VFS. Environ Toxicol Chem 2021;40:2615-2628. © 2021 SETAC.

Effects of a novel fungicide benzovindiflupyr in Eisenia fetida: Evaluation through different levels of biological organization.

Although benzovindiflupyr (BZF), which is a novel succinate dehydrogenase inhibitor fungicide, has considerable application potential worldwide, its extensive use is toxic to non-targeted soil organisms. Therefore, this study aimed to evaluate the acute and subchronic toxicity of BZF to earthworms (Eisenia fetida). The acute toxicity of BZF to adult and larval earthworms was measured, as indicated by the following LC50 values obtained after 14 days of exposure: 416 mg/kg for adult earthworms and 341 mg/kg for juveniles. Subchronic toxicity tests were conducted using only adult earthworms. The earthworms' weight gain was slower on days 14 and 28 after commencing the BZF T100 treatment (50 mg/kg of soil). Following 14 days of BZF exposure, enzymes and gene expressions associated with the mitochondrial respiratory chain and energy metabolism were activated to some extent, and the reactive oxygen species level and malondialdehyde content also increased. Antioxidant and detoxifying enzymes and metallothionein gene, Heat shock protein 70 gene associated with resistance to oxidative damage were also activated to varying degrees. Increased BZF concentrations corresponded to increased genotoxicity. Integrated biological response (IBR) values were calculated at the biochemical and molecular levels to show increased toxicity with increased BZF concentration. Although a series of biomarkers changes occurred after initiating BZF treatment, these changes were all likely to have been resisted by the earthworms' own antioxidant defense system and only showed phenotypic (weight-related) changes with treatments of 50 mg/kg. In conclusion, reasonable levels of BZF application may have little impact on earthworms. Our findings provide insights on the toxic effects of BZF on earthworms and may prove useful for risk assessments relating to BZF's impacts on soil ecosystems.

Favorable Bioactivity of the SDHI Fungicide Benzovindiflupyr Against Sclerotinia sclerotiorum Mycelial Growth, Sclerotial Production, and Myceliogenic and Carpogenic Germination of Sclerotia.

Sclerotinia sclerotiorum, which can cause Sclerotinia stem rot, is a prevalent plant pathogen. This study aims to evaluate the application potential of benzovindiflupyr, a new generation of succinate dehydrogenase inhibitor (SDHI), against S. sclerotiorum. In our study, 181 isolates collected from different crops (including eggplant [n = 34], cucumber [n = 27], tomato [n = 29], pepper [n = 35], pumpkin [n = 32], and kidney bean [n = 25]) in China were used to establish baseline sensitivity to benzovindiflupyr. The frequency distribution of the 50% effective concentration (EC50) values of benzovindiflupyr was a unimodal curve, with mean EC50 values of 0.0260 ± 0.011 µg/ml, and no significant differences in mean EC50 existed among the various crops (P > 0.99). Benzovindiflupyr can effectively inhibit mycelial growth, sclerotial production, sclerotial shape, and myceliogenic and carpogenic germination of the sclerotia of S. sclerotiorum. In addition, benzovindiflupyr showed good systemic translocation in eggplant. Using benzovindiflupyr at 100 µg/ml yielded efficacies of 71.3 and 80.5% for transverse activity and cross-layer activity, respectively, which were higher than those of acropetal and basipetal treatments (43.6 and 44.7%, respectively). Greenhouse experiments were then carried out at two experimental sites for verification. Applying benzovindiflupyr at 200 g a.i. ha-1 significantly reduced the disease incidence and severity of Sclerotinia stem rot. Overall, the results demonstrated that benzovindiflupyr is a potential alternative product to control Sclerotinia stem rot.

Baseline sensitivity of Bipolaris maydis to the novel succinate dehydrogenase inhibitor benzovindiflupyr and its efficacy.

Benzovindiflupyr is a novel member of succinate dehydrogenase inhibitor (SDHI) fungicides. The filamentous fungus Bipolaris maydis Nisik. et Miyake was the causal agent of southern corn leaf blight (SCLB). Here, baseline sensitivity of B. maydis to benzovindiflupyr was established by mycelial growth and conidium germination methods using 96 B. maydis isolates collected from various places of Jiangsu Province of China, and EC50 values ranged from 0.0321 to 0.9149 µg/ml with the mean value of 0.3446 (±0.2248) µg/ml for mycelial growth, and 0.1864 to 0.964 µg/ml with the mean value of 0.5060 (±0.2094) µg/ml for conidium germination respectively. Treated with benzovindiflupyr, the distribution of nuclei and septum of hyphae did not change, but hyphae of offshoot and conidial production of B. maydis decreased significantly, the cell membrane permeability increased. The result of transmission electron microscope showed that the cross section of hypha was out of shape, the cell wall became thin and sparse, the cell membrane were distinctly damaged, organelles dissolved and vacuolated, and the cell nearly broke up. The results suggested that benzovindiflupyr had strong activity against mycelial growth and conidial production of B. maydis by damaging cell wall, membrane and organelles. The protective and curative activity assays for benzovindiflupyr indicated that benzovindiflupyr exhibited excellent suppression of B. maydis development on detached corn leaves. In protective activity assay with application of benzovindiflupyr at 10 µg/ml, the control efficacy reached to 100%. In curative activity assay with application of benzovindiflupyr at 50 µg/ml, the control efficacy reached to 90.72%. This is the first report of baseline sensitivity of B. maydis to benzovindiflupyr and its biological activity against B. maydis. It is recommended that benzovindiflupyr is a excellent candidate for controlling SCLB.

High-Efficiency Control of Gray Mold by the Novel SDHI Fungicide Benzovindiflupyr Combined with a Reasonable Application Approach of Dipping Flower.

In this study, a novel succinate dehydrogenase inhibitor (SDHI) fungicide benzovindiflupyr was found to have strong inhibitory activity against gray mold caused by Botrytis cinerea. The sensitivity of B. cinerea to benzovindiflupyr was determined by testing 103 pathogen isolates with mean values of 2.15 ± 0.19 mg L-1 and 0.89 ± 0.14 mg L-1 for mycelial growth and spore germination inhibition, respectively. Furthermore, benzovindiflupyr had excellent long-lasting protective activity. Unfortunately, there were positive correlations between benzovindiflupyr and boscalid ( r = 0.3, P = 0.04) and between benzovindiflupyr and isopyrazam ( r = 0.31, P = 0.04). In the field, cucumber flowers are susceptible to infection by B. cinerea. Benzovindiflupyr applied at 20 mg L-1 by dipping flowers could successfully control cucumber gray mold, with the benzovindiflupyr dose of dipping flower application less than 1% of that of spraying application. Benzovindiflupyr combined with dipping flower application showed significant control of gray mold.