Increasing Cyetpyrafen Spray Volume and Ozone Spray Improves the Control Effects against Two-Spotted Spider Mite (Tetranychus urticae) in Strawberries.

The two-spotted spider mite (Tetranychus urticae) is a constant threat to greenhouse strawberry production. The application of synthetic acaricides is the main method of controlling T. urticae. However, resistance development to traditional acaricides reduces their efficacy and eventually leads to control failure. It is important for strawberry growers to look for new acaricides and application technologies that can limit the harmfulness of T. urticae in environmentally friendly ways. In the current study, laboratory toxicity tests and field trials were performed to screen high-efficiency acaricides, and then application technologies were improved to enhance the management of T. urticae. In the laboratory toxicity tests, the results showed that the LC50 (median lethal concentration) value of cyetpyrafen, cyenopyrafen, cyflumetofen, bifenazate, abamectin, azocyclotin, pyridaben, spirodiclofen, and etoxazole against adult T. urticae was 0.226, 0.240, 0.415, 3.583, 5.531, 25.58, 39.69, 140.3, and 267.7 mg/L, respectively. In addition, the LC50 value of the nine acaricides against eggs of T. urticae was 0.082, 0.097, 0.931, 18.56, 25.52, 45.61, 36.32, 1.954, and 0.040 mg/L, respectively. The field trial results showed that the best control effect was obtained in cyetpyrafen at 300 mL/ha treatment. Cyetpyrafen was chosen for further application technology tests. In the spray volume tests, the results showed that increasing the spray volume from 900 to 1050 L/ha significantly improved the control of T. urticae. In addition, the results from the spray instrument tests demonstrated that the control effects on T. urticae in the ozone spray treatments were significantly higher than those of the conventional and electrostatic sprays 1 and 3 days after treatment (DAT). Therefore, this study suggested that cyetpyrafen effectively controlled T. urticae both in the laboratory tests and in the field trials. Increasing the spray volume and application of ozone spray significantly improved T. urticae management.

A systemic study of cyenopyrafen in strawberry cultivation system: Efficacy, residue behavior, and impact on honeybees (Apis mellifera L.).

The pesticide application method is one of the important factors affecting its effectiveness and residues, and the risk of pesticides to non-target organisms. To elucidate the effect of application methods on the efficacy and residue of cyenopyrafen, and the toxic effects on pollinators honeybees in strawberry cultivation, the efficacy and residual behavior of cyenopyrafen were investigated using foliar spray and backward leaf spray in field trials. The results showed that the initial deposition of cyenopyrafen using backward leaf spray on target leaves reached 5.06-9.81 mg/kg at the dose of 67.5-101.25 g a.i./ha, which was higher than that using foliar spray (2.62-3.71 mg/kg). The half-lives of cyenopyrafen in leaves for foliar and backward leaf spray was 2.3-3.3 and 5.3-5.9 d, respectively. The residues (10 d) of cyenopyrafen in leaves after backward leaf spray was 1.41-3.02 mg/kg, which was higher than that after foliar spraying (0.25-0.37 mg/kg). It is the main reason for the better efficacy after backward leaf spray. However, the residues (10 d) in strawberry after backward leaf spray and foliar spray was 0.04-0.10 and < 0.01 mg/kg, which were well below the established maximum residue levels of cyenopyrafen in Japan and South Korea for food safety. To further investigate the effects of cyenopyrafen residues after backward leaf spray application on pollinator honeybees, sublethal effects of cyenopyrafen on honeybees were studied. The results indicated a significant inhibition in the detoxification metabolic enzymes of honeybees under continuous exposure of cyenopyrafen (0.54 and 5.4 mg/L) over 8 d. The cyenopyrafen exposure also alters the composition of honeybee gut microbiota, such as increasing the relative abundance of Rhizobiales and decreasing the relative abundance of Acetobacterales. The comprehensive data on cyenopyrafen provide basic theoretical for environmental and ecological risk assessment, while backward leaf spray proved to be effective and safe for strawberry cultivation.

Novel Acaricidal Silico-Containing Pyrazolyl Acrylonitrile Derivatives Identified through Rational Carbon-Silicon Bioisosteric Replacement Strategy.

The identification of novel pyrazolyl acrylonitrile acaricides with improved properties is of great value for the control of phytophagous mites. A series of innovative silicon-containing pyrazolyl acrylonitriles were rationally designed by applying a bioisosteric carbon-silicon replacement strategy and prepared based on novel synthetic methodology. As a result of our research, we discovered compound A25 which possesses outstanding acaricidal activity. With an LC50 value of 0.062 mg/L, compound A25 was found to be 2.3-fold and 1.9-fold more potent than the commercial acaricides cyenopyrafen and cyetpyrafen, respectively. Enzymatic inhibitory assay indicated that the active principle M1 of compound A25 possesses an IC50 value of 2.32 µM against Tetranychus cinnabarinus SDH, which was about twofold superior compared to the active metabolites of cyenopyrafen (IC50 = 4.72 µM). Molecular docking study showed that the active metabolites 2 and 3 and their corresponding silicon counterparts form H-bonds and cation-π interaction with the residues of Trp165, Tyr433, and Arg279.

The dissipation behavior, household processing factor and risk assessment for cyenopyrafen residues in strawberry and mandarin fruits.

A modified QuEChERS method for determining cyenopyrafen in strawberries, mandarins and their processed products was established with a good linearity (R2 > 0.9981), accuracy (recoveries of 83% to 111%) and precision (relative standard deviations of 0.9% to 14%). The limit of quantification (LOQ) was 0.01 mg/kg. Field results showed that the half-lives of cyenopyrafen were 6.8 and 11.8 d in strawberry and mandarin respectively, and that the final residues were within established maximum residue limits (MRLs). The household processing factors (PFs) for cyenopyrafen residues in strawberry and mandarin fruits were also studied: residues increased in strawberry jam (PF 1.51) and mandarin juice (1.31) but decreased in strawberries (0.58) and mandarin pulp (<0.17) after washing and peeling, respectively. A risk assessment showed that the risk from long-term dietary exposures to cyenopyrafen was 73.73%, indicating that consuming these products was unlikely to present a public health concern.

Decline pattern and risk assessment of cyenopyrafen in different varieties of Asian pear using liquid chromatography and tandem mass spectrometry.

The dissipation pattern of a commercial cyenopyrafen formulation sprayed at the recommended dose on Asian pears (two different species) grown at two different sites was investigated using liquid chromatography-ultraviolet detection. Samples collected randomly over 14 days were extracted using acetone, partitioned using n-hexane/dichloromethane (8/2, v/v), and purified using a Florisil solidphase extraction cartridge. The residues in field-incurred samples were confirmed via liquid chromatography-tandem mass spectrometry. The method was validated in terms of excellent linearity in the solvent (R 2=1); moreover, satisfactory recoveries (89.0-107.3%) were obtained at three fortification levels with a relative standard deviation (RSD)≤5.0% and the limits of detection and quantification of 0.0033 and 0.01 mg/kg, respectively. Although the residual levels at both sites were lower than the maximum residue limit (MRL=1 mg/kg), the dissipation at Site 2 was faster than that at Site 1. Consequently, the half-life (t1/2) in Site 2 (5.2 d) was shorter than that in Site 1 (9.8 d). Risk assessment at zero days showed acceptable daily intakes (%) of 27.25% and 24.52% at Sites 1 and 2, respectively, indicating that these fruit species are safe for consumption.

Functional characterization of the Tetranychus urticae CYP392A11, a cytochrome P450 that hydroxylates the METI acaricides cyenopyrafen and fenpyroximate.

Cyenopyrafen is a Mitochondrial Electron Transport Inhibitor (METI) acaricide with a novel mode of action at complex II, which has been recently developed for the control of the spider mite Tetranychus urticae, a pest of eminent importance globally. However, some populations of T. urticae are cross-resistant to this molecule, and cyenopyrafen resistance can be readily selected in the lab. The cytochrome P450s genes CYP392A11 and CYP392A12 have been strongly associated with the phenotype. We expressed the CYP392A11 and the CYP392A12 genes with T. urticae cytochrome P450 reductase (CPR) in Escherichia coli. CYP392A12 was expressed predominately as an inactive form, witnessed by a peak at P420, despite optimization efforts on expression conditions. However, expression of CYP392A11 produced a functional enzyme, with high activity and preference for the substrates Luciferin-ME EGE and ethoxycoumarin. CYP392A11 catalyses the conversion of cyenopyrafen to a hydroxylated analogue (kcat = 2.37 pmol/min/pmol P450), as well as the hydroxylation of fenpyroximate (kcat = 1.85 pmol/min/pmol P450). In addition, transgenic expression of CYP392A11 in Drosophila melanogaster, in conjunction with TuCPR, confers significant levels of fenpyroximate resistance. The overexpression of CYP392A11 in multi-resistant T. urticae strains, not previously exposed to cyenopyrafen, which had been indicated by microarray studies, was confirmed by qPCR, and it was correlated with significant levels of cyenopyrafen and fenpyroximate cross-resistance. The implications of our findings for insecticide resistance management strategies are discussed.

The economic importance of acaricides in the control of phytophagous mites and an update on recent acaricide mode of action research.

Acaricides are one of the cornerstones of an efficient control program for phytophagous mites. An analysis of the global acaricide market reveals that spider mites such as Tetranychus urticae, Panonychus citri and Panonychus ulmi are by far the most economically important species, representing more than 80% of the market. Other relevant mite groups are false spider mites (mainly Brevipalpus), rust and gall mites and tarsonemid mites. Acaricides are most frequently used in vegetables and fruits (74% of the market), including grape vines and citrus. However, their use is increasing in major crops where spider mites are becoming more important, such as soybean, cotton and corn. As revealed by a detailed case study of the Japanese market, major shifts in acaricide use are partially driven by resistance development and the commercial availability of compounds with novel mode of action. The importance of the latter cannot be underestimated, although some compounds are successfully used for more than 30 years. A review of recent developments in mode of action research is presented, as such knowledge is important for devising resistance management programs. This includes spirocyclic keto-enols as inhibitors of acetyl-CoA carboxylase, the carbazate bifenazate as a mitochondrial complex III inhibitor, a novel class of complex II inhibitors, and the mite growth inhibitors hexythiazox, clofentezine and etoxazole that interact with chitin synthase I.

Cross-resistance between cyenopyrafen and pyridaben in the twospotted spider mite Tetranychus urticae (Acari: Tetranychidae).

BACKGROUND: Cyenopyrafen is an inhibitor of complex II of the mitochondrial electron transport chain. It has a molecular structure that shares some common features with frequently used complex I inhibitors such as pyridaben. To evaluate whether this similarity in structure poses a cross-resistance risk that might complicate resistance management, we selected for pyridaben and cyenopyrafen resistance in the laboratory and characterized resistance. RESULTS: The selection for cyenopyrafen conferred cross-resistance to pyridaben and vice versa. Resistance towards these both acaricides was incompletely dominant in adult females. However, in eggs maternal effects were observed in pyridaben resistance, but not in the cyenopyrafen-resistance (completely dominant). In the cyenopyrafen resistant strain, the LC50 of eggs remained lower than the commercially recommended concentration. The common detoxification mechanisms by cytochrome P450 was involved in resistance to these acaricides. Carboxyl esterases were also involved in cyenopyrafen resistance as a major factor. CONCLUSIONS: Although cross-resistance suggests that pyridaben resistance would confer cyenopyrafen cross-resistance, susceptibility in eggs functions to delay the development of cyenopyrafen resistance.

Cross-resistance risk of the novel complex II inhibitors cyenopyrafen and cyflumetofen in resistant strains of the two-spotted spider mite Tetranychus urticae.

BACKGROUND: Cyflumetofen and cyenopyrafen are novel acaricides acting as complex II inhibitors. This new mode of action is extremely useful for devising efficient resistance management strategies for mite control. The authors determined the cross-resistance risk of both compounds, using a collection of well-characterised resistant strains of Tetranychus urticae, and also selected for cyflumetofen resistance in the laboratory. RESULTS: Cross-resistance to cyflumetofen and cyenopyrafen was detected in field strains, with LC50 values exceeding the registered field dose. Synergism experiments suggested that P450 monooxygenases are involved in resistance, and that the activation mechanism of the two compounds most likely differs. Laboratory selection with cyflumetofen resulted in a highly resistant T. urticae strain that displayed negative cross-resistance to cyenopyrafen. CONCLUSIONS: The cross-resistance risk of cyflumetofen and cyenopyrafen documented in this study needs to be integrated in resistance management strategies, especially in regions or crops with a history of frequent acaricide applications, in order to safeguard the efficacy of these compounds with a valuable new mode of action.