Diamide Insecticides Research Articles

Molecular characterization and expression patterns of a ryanodine receptor in soybean looper, Chrysodeixis includens.

Diamide insecticides, such as chlorantraniliprole, have been widely used to control insect pests by targeting the insect ryanodine receptor (RyR). Due to the efficacious insecticidal activity of diamides, as well as an increasing number of resistance cases, the molecular structure of RyR has been studied in many economically important insects. However, no research has been conducted on diamide resistance and RyR in the soybean looper, Chrysodeixis includens, a significant crop pest. In this study, we found moderate resistance to chlorantraniliprole in a field population from Puerto Rico and sequenced the full-length cDNA of the C. includens RyR gene, which encodes a 5124 amino acid-long protein. Genomic analysis revealed that the CincRyR gene consists of 113 exons, one of the largest exon numbers reported for RyR. Alternative splicing sites were detected in the cytosolic region. The protein sequence showed high similarity to other noctuid RyRs. Conserved structural features included the selectivity filter motif critical for ryanodine binding and ion conduction, as well as various domains involved in ion transport. Two mutation sites associated with diamide resistance in other insects were screened but not found in the Puerto Rico field populations or in the susceptible lab strain. Gene expression analysis indicated high expression of RyR in the third instar larval stage, particularly in muscle-containing tissues. Furthermore, exposure to a sublethal dose of chlorantraniliprole reduced RyR expression levels after 96 h. This study provides a molecular basis for understanding RyR structure and sheds light on potential mechanisms of diamide resistance in C. includens.

Exaptation of I4760M mutation in ryanodine receptor of Spodoptera exigua (Lepidoptera: Noctuidae): Lessons from museum and field samples

Since 2007, diamide insecticides have been widely used in Korea to control various types of lepidopteran pests including Spodoptera exigua. For nearly a decade, diamide resistance in field populations of S. exigua across 18 localities has been monitored using bioassays. Despite their short history of use, resistance to diamide insecticides has emerged. Based on the LC50 values, some field populations showed a higher level of resistance to chlorantraniliprole, a diamide insecticide, compared to that of the susceptible strain, although regional and temporal variations were observed. To investigate resistance at a molecular level, we examined three mutations (Y4701C, I4790M, and G4946E) in the ryanodine receptor (RyR), which is the primary mechanism underlying diamide insecticide resistance. DNA sequencing showed that only the I4790M mutation was found in most field populations. As resistance levels varied significantly despite the uniform presence of the I4790M mutation, we considered the presence of another resistance factor. Further, the I4790M mutation was also found in S. exigua specimens collected prior to the commercialization of diamide insecticides in Korea as well as in other countries, such as the USA. This finding led us to hypothesize that the I4790M mutation were predisposed in field populations owing to selection factors other than diamide use. For further clarification, we conducted whole-genome sequencing of S. exigua (449.83 Mb) and re-sequencing of 18 individual whole genomes. However, no additional non-synonymous mutations were detected in the RyR-coding region. Therefore, we concluded that the high level of diamide insecticide resistance in Korean S. exigua is not caused by mutations at the target site, RyR, but is attributed to other factors that need to be investigated in future studies.

Characterization of Six Diamide Insecticides on Ryanodine Receptor: Resistance and Species Selectivity.

Ryanodine receptor (RyR) has been used as an insecticide target to control many destructive agricultural pests. The effectiveness of these insecticides has been limited by the spread of resistance mutations identified in pest RyRs, but the detailed molecular impacts of the individual mutations on the activity of different diamide compounds have not been fully explored. We created five HEK293 cell lines stably expressing wild type rabbit RyR1, wild type Spodoptera frugiperda RyR (Sf RyR), or Sf RyR carrying different resistance mutations, including G4891E, G4891E/I4734M, and Y4867F, respectively. R-CEPIA1er, a genetically encoded fluorescent protein, was also introduced in these cell lines to report the Ca2+ concentration in the endoplasmic reticulum. We systematically characterized the activities of six commercial diamide insecticides against different RyRs using the time-lapse fluorescence assay. Among them, cyantraniliprole (CYAN) displayed the highest activity against all three resistant Sf RyRs. The good performance of CYAN was confirmed by the toxicity assay using gene-edited Drosophila expressing the mutant RyRs, in which CYAN showed the lowest LD50 value for the double resistant mutant. In addition, we compared their acitivty between mammalian and insect RyRs and found that flubendiamide has the best insect-selectivity. The mechanism of the anti-resistance property and selectivity of the compounds was proposed based on the structural models generated by homology modeling and molecular docking. Our findings provide insights into the mechanism of insect resistance and guidance for developing effective RyR agonists that can selectively target resistant pests.

Comparative analyses of the effects of sublethal doses of emamectin benzoate and tetrachlorantraniliprole on the gut microbiota of Spodoptera frugiperda (Lepidoptera: Noctuidae).

Spodoptera frugiperda (J. E. Smith) is an important invasive pest that poses a serious threat to global crop production. Both emamectin benzoate (EB) and diamide insecticides are effective insecticides used to protect against S. frugiperda. Here, 16S rRNA sequencing was used to characterize the gut microbiota in S. frugiperda larvae exposed to EB or tetrachlorantraniliprole (TE). Firmicutes and Proteobacteria were found to be the dominant bacterial phyla present in the intestines of S. frugiperda. Following insecticide treatment, larvae were enriched for species involved in the process of insecticide degradation. High-level alpha and beta diversity indices suggested that exposure to TE and EB significantly altered the composition and diversity of the gastrointestinal microbiota in S. frugiperda. At 24 h post-EB treatment, Burkholderia-Caballeronia-Paraburkholderia abundance was significantly increased relative to the control group, with significant increases in Stenotrophobacter, Nitrospira, Blastocatella, Sulfurifustis, and Flavobacterium also being evident in these larvae. These microbes may play a role in the degradation or detoxification of EB and TE, although further work will be needed to explore the mechanisms underlying such activity. Overall, these findings will serve as a theoretical foundation for subsequent studies of the relationship between the gut microbiota and insecticide resistance in S. frugiperda (J. E. Smith) (Lepidoptera: Noctuidae).

Overexpression of the UDP-glycosyltransferase UGT34A23 confers resistance to the diamide insecticide chlorantraniliprole in the tomato leafminer, Tuta absoluta

The tomato leafminer, Tuta absoluta, is an invasive crop pest that has evolved resistance to many of the insecticides used for its control. To facilitate the investigation of the underpinning mechanisms of resistance in this species we generated a contiguous genome assembly using long-read sequencing data. We leveraged this genomic resource to investigate the genetic basis of resistance to the diamide insecticide chlorantraniliprole in Spanish strains of T. absoluta that exhibit high levels of resistance to this insecticide. Transcriptomic analyses revealed that, in these strains, resistance is not associated with previously reported target-site mutations in the diamide target-site, the ryanodine receptor, but rather is associated with the marked overexpression (20- to >100-fold) of a gene encoding a UDP-glycosyltransferase (UGT). Functional expression of this UGT, UGT34A23, via ectopic expression in Drosophila melanogaster demonstrated that it confers strong and significant resistance in vivo. The genomic resources generated in this study provide a powerful resource for further research on T. absoluta. Our findings on the mechanisms underpinning resistance to chlorantraniliprole will inform the development of sustainable management strategies for this important pest.

Toxicity and effects on anuran tadpole metamorphosis of the anthranilic diamide insecticides chlorantraniliprole and cyantraniliprole

ABSTRACT The present study examined the acute and chronic toxicity attributed to commercial formulations of the anthranilic diamide insecticides chlorantraniliprole (CHLO) and cyantraniliprole (CYAN) on the neotropical amphibian species Rhinella arenarum, Rhinella fernandezae and Scinax granulatus. The median lethal concentrations obtained after 96 hr exposure (96 hr-LC50) were generally greater than 100 mg/L, except for stage 25 S. Granulatus, which were the most sensitive animals tested with a 96 hr-LC50 value of 46.78 mg/L. In subchronic exposures of R. arenarum, the 21day-LC50 were 151.4 mg/L for CHLO and >160 mg/L for CYAN, the weight gain of the tadpoles during this period not being markedly affected in both cases. Finally, when tadpoles of R. arenarum were exposed to CHLO throughout the metamorphic process, an inverted U-shaped non-monotonic dose-response relationship was observed between exposure concentrations and both % of individuals transiting between stage 39 and 42 and the time required to accomplish this. Data obtained raise the hypothesis of an effect of CHLO on the hypothalamic-pituitary-thyroid (HPT) axis, either directly or through an interaction with the stress-hormone system, as metamorphic progression from stage 39 to S42 occurs under the strict control of thyroid hormones. These observations are important as the anthranilic diamide insecticides are not currently known as endocrine disruptors. Further investigations are needed to clarify the pathways leading to these effects and examine whether environmentally-relevant aquatic concentrations of anthranilic diamides might be impacting amphibian populations in the wild.

The Selective Detection of Cyantraniliprole Insecticides Using Molecularly Imprinted Polymers Coupled with an Acetylcholinesterase Inhibition-Based Biosensor.

Cyantraniliprole is one of the anthranilic diamide insecticides widely used in the agriculture sector. Due to its low toxicity and relatively fast degradation, there is need for a sensitive determination method for its residues. Nowadays, there is growing interest in the development of enzyme-based biosensors. The major drawback is the non-specific binding of many insecticides to the enzyme. This work employs Molecularly imprinted polymers (MIPs) to increase enzyme specificity and eliminate the organic solvent effect on the enzyme activity. The synthesized Cyan-Molecularly imprinted polymers (Cyan-MIP) possesses high affinity and selectivity toward cyantraniliprole. Acetylcholinesterase assay characteristics including enzyme concentration, substrate concentration, DTNB concentration, and acetonitrile concentration were optimized. Under optimal experimental conditions, the developed MIP-Acetylcholinesterase (MIP-AchE) inhibition-based sensor provides better precision than the AchE inhibition-based sensor with a wide linear range (15-50 ppm), limit of detection (LOD) 4.1 ppm, and limit of quantitation (LOQ) 12.6 ppm. The sensor was successfully applied for cyantraniliprole determination in spiked melon, giving satisfactory recoveries.

Monitoring and characterization of field‐evolved resistance to diamide insecticides in Spodoptera litura collected from eastern China

AbstractSpodoptera litura is a notoriously destructive pest, which infests horticultural plants and economically important crops worldwide. Several generations of diamide insecticides have been used to combat S. litura throughout the world, causing various levels of resistance to this class of insecticides among wild S. litura populations. In this work, we monitored resistance to five diamide insecticides among 10 S. litura populations sampled from eastern China in 2022. In comparison with Lab‐S the susceptible strain, five of the tested populations displayed high susceptibility to three of the five diamide insecticides (cyantraniliprole, broflanilide and tetraniliprole). In contrast, there was significant resistance to the other two tested diamide insecticides, flubendiamide and chlorantraniliprole, in seven of the field‐collected populations. The Shanghai (SH) population showed high resistance to chlorantraniliprole (77.0‐fold), flubendiamide (25.5‐fold) and cyantraniliprole (27.8‐fold), respectively, than the strain of Lab‐S. Results of synergism tests in the SH population revealed that diethyl maleate (DEM) and piperonyl butoxide (PBO) greatly inhibited chlorantraniliprole resistance. The progeny of reciprocal crosses between the Lab‐S and SH populations presented dominance degrees of F1 progenies, suggesting that the chlorantraniliprole resistance was autosomal and incompletely recessive in SH. Above results of the current work offer useful knowledge for future formulation of field pest management strategies, allowing farmers to delay the development of resistance to diamide insecticides and thus promote sustainable management of S. litura.

The ryanodine receptor mutation I4728M confers moderate-level resistance to diamide insecticides in Spodoptera litura.

The common cutworm, Spodoptera litura (Fabricius), is one of the most widespread and destructive polyphagous pests in tropical and subtropical Asia. S. litura has evolved resistance to different insecticides, including diamide insecticides. Here, we identified a ryanodine receptor (RyR) mutation (I4728M) associated with target site resistance to diamides in a field-collected population of S. litura. The contribution of this mutation to diamide resistance was investigated through establishing a near-isogenic resistant strain of S. litura. The ND21 population of S. litura, collected from Ningde, Fujian province of China in 2021, exhibited 130.6-fold resistance to chlorantraniliprole compared to the susceptible NJ-S strain. S. litura RyR mutation I4728M, corresponding to Plutella xylostella RyR I4790M, was identified in the ND21 population. SlRyR I4728M mutation of ND21 was introgressed into a susceptible background strain (NJ-S) with marker-assisted backcrossing. The introgressed strain named ND21-R, which was homozygous for the mutant 4728M allele, shared about 94% of the genetic background with the NJ-S strain. ND21-R strain showed moderate levels of resistance to two anthranilic diamides (19.1-fold to chlorantraniliprole, 19.7-fold to cyantraniliprole) and the phthalic diamide flubendiamide (23.4-fold). Genetic analysis showed that chlorantraniliprole resistance was autosomal, incompletely recessive and tightly linked with SlRyR I4728M mutation in the introgressed ND21-R strain of S. litura. Identification of the I4728M mutation and its contribution to diamide resistance in S. litura will help develop allelic discrimination assays for resistance monitoring and guide resistance management practices for diamides in S. litura. © 2023 Society of Chemical Industry.

Diamide insecticides targeting insect ryanodine receptors: Mechanism and application prospect

As a Lepidoptera pest, Spodoptera frugiperda has become one of the major migratory pests causing significant damage to crops. It should prevent and control Spodoptera frugiperda with strong reproductive ability, adaptability, and migration ability, and reduce economic losses as much as possible. Chemical insecticides are mainly used in the emergency control of Spodoptera frugiperda. Diamide insecticide is a kind of pesticide that specifically targets the ryanodine receptor of Lepidopteran pests, which makes it safe, effective, targeted, and low toxicity to mammals. So, it is one of the most concerned and fastest-growing pesticide products after neonicotinoid pesticides. Intracellular Ca2+ concentration can be regulated by ryanodine receptors, and the continuous release of Ca2+ eventually leads to the death of pests and achieve the insecticidal effect. This review introduces in detail diamide insecticides that mainly play roles in stomach toxicity, as well as its specific target-ryanodine receptor, and analyzes how the diamide insecticide acts on the ryanodine receptor and how its mechanism of action can provide a theoretical basis for the rational use of highly effective insecticides and solve the resistance problem. Moreover, we also propose several recommendations for reducing resistance to diamide insecticides, and provide a reference for chemical control and resistance studies of Spodoptera frugiperda, which has broad development prospects in today's increasingly concerned about the ecological environment and advocating green environmental protection.

Evaluation of the Two Typical Diamide Insecticide-Induced Oxidative Damages and the Molecular Mechanism Underlying Their Toxicity in Triticum aestivum.

As the typical representatives of diamide insecticides, excessive exposure to flubendiamide and chlorantraniliprole for plants may inevitably pose threats to plant growth and food safety. However, the underlying toxic mechanisms remain unclear. Here, glutathione S-transferase Phi1 from Triticum aestivum was employed as the biomarker to assess oxidative damages. First, flubendiamide displayed much stronger binding affinity with TaGSTF1 than chlorantraniliprole in consistent with molecular docking results, and flubendiamide also exerted more evident effects on the structure of TaGSTF1. Then, glutathione S-transferase activities of TaGSTF1 declined after interaction with these two insecticides, especially for flubendiamide with more hazardous influence. At last, the adverse impacts on the germination and growth of wheat seedlings were further evaluated with more apparent inhibition of flubendiamide. Hence, this study may illustrate the detailed binding mechanisms of TaGSTF1 with these two typical insecticides, evaluate the destructive impacts on plant growth, and further assess the threat to agriculture.

Design, Synthesis, and Properties of Silicon-Containing meta-Diamide Insecticides.

Silicon-containing compounds are sporadically used in crop protection and drug discovery and have demonstrated to increase the biological efficacy as well as to reduce toxicity, improve physicochemical properties, and favorably impact the environmental profile. As part of our research, we have investigated the application of bioisosteric silicon replacements in meta-diamide insecticides and studied the biological activity and molecular properties of the corresponding novel compounds. At all meaningful structural elements of the meta-diamides, silicon-containing substituents were introduced and synthetic methodology was developed for their syntheses. As the most promising compound, silicon-containing meta-diamide II-18 emerged, which exhibits a very low LC50 value of 2.00 mg/L against Mythimna separata and compares well to the reference compounds 28 (LC50 = 0.17 mg/L) and II-20 (LC50 = 0.27 mg/L). Our research on silicon-containing crop protection compounds once again confirmed that the biological activity can be beneficially affected by the insertion of silicone substituents and that the introduction of well-chosen silicone motifs is an excellent strategy for agrochemical research.

Flubendiamide induced genetic and cellular damages directly influence the life cycle of the oriental leaf worm, Spodoptera litura

Indiscriminate uses of insecticide greatly damage the environment as well as non-target organisms. Thus, multiple levels of bioassays can help better management of our environment. Flubendiamide is a phthalic acid diamide insecticide that ceases the function of insect muscle leading to paralysis and death. Here, we aimed to explore the effects of Flubendiamide on the life cycle of Spodoptera litura vis-a-vis the mode of action. Fourth instar larvae of the same age (120 ± 2 h) and size were fed with different concentrations (20–80 μg/mL) of Flubendiamide for 12–72 h. We performed a pharmacokinetics study, different biochemical assays, p450, Ecdysone receptor (EcR) and other genes expression analyses by Real-Time PCR and gross damages by Dye exclusion assay and histopathology. Our results demonstrate that the mean concentration of Flubendiamide after 48 h is 9.907 μg/mL and (i) altered the molting, metamorphosis, and reproduction at 80 μg/mL (24 h) (ii) increases all oxidative stress parameters (ROS/RNS, MDA, 8OHdG), decreases oxidative defense mechanisms (SOD, CAT, GST) at 80 μg/mL (48 h) and p450 in a time and concentration-dependent manner, (iii) activates CncC/Maf apoptotic pathways at 80 μg/mL concentration at 24 h while the expression declined from 48 h onwards, (iii) downregulates the EcR expression in a time and concentration-dependent manner, which might be responsible for disturbed molting, metamorphosis, and reproduction, and (iv) increase the expression of apoptotic genes (Caspase 1, −3, and − 5), in time and concentration-dependent manner causing gross morphological and histological damages. In conclusion, indiscriminate use of this insecticide can affect the ecosystem and have the capacity to cause multiple hazardous effects on experimental organisms. Thus, it warrants further investigations to improve and optimize the integrated pest management packages, including Flubendiamide for better management.

Toxicity and Sublethal Effects of Diamide Insecticides on Key Non-Target Natural Predators, the Larvae of Coccinella septempunctata L. (Coleoptera: Coccinellidae).

Coccinella septempunctata (ladybird) is an extremely important natural predator that feeds on aphids. An assessment of the toxicity of pesticides on environmental organisms is an essential component of Integrated Pest Management (IPM) strategies. This study evaluated diamide insecticides' toxicity at lethal and 30% lethal doses (LR30) against C. septempunctata larvae. The pre-imaginal median lethal doses (LR50) of chlorantraniliprole 10% SC, tetrachlorantraniliprole 10% SC, and broflanilide 10% SC were calculated to be 42.078, 289.516, and 0.0943 g active ingredient (a.i.)/ha, respectively. The mortality tests demonstrated that chlorantraniliprole and tetrachlorantraniliprole are comparatively less toxic to C. septempunctata than broflanilide, which were detected to be highly toxic to C. septempunctata. The mortality rates of the groups treated with the three diamide insecticides tended to stabilize after 96 h, extending to the pre-imaginal stage. Furthermore, when compared to broflanilide, which had a much higher potential risk, the hazard quotient (HQ) values indicated that chlorantraniliprole and tetrachlorantraniliprole have a lower risk potential for C. septempunctata in farmland and off farmland. The LR30 dose induces abnormalities in the development phase 4th-instar larvae weight, pupal weight, and adult weight of treated C. septempunctata. The study emphasizes the importance of assessing the adverse effects of diamide insecticides on natural predator species that serve as biological control agents in agricultural IPM strategies.

Bio-efficacy of chlorantraniliprole 0.4% gr against Cnaphalocrocis medinalis and Scirpophaga incertulas in rice (Oryza sativa)

With the widespread adoption of high yielding rice (Oryza sativa L.) varieties and accompanying changes in the cultural practices, rice leaf folder [Cnaphalocrocis medinalis (Guenee)] and yellow stem borer [Scirpophaga incertulas (Walker)] has attained the status of major pests and influence the yield if proper timely management practices are not adopted. For effective management of these pests, identification of new molecules with novel mode of action, low toxicity to non-target insects and environmental safety is required for sustainable IPM. Chlorantraniliprole 0.4% gr (Ferterra 0.4% gr) is an anthranillic diamide insecticide in granular form for the control of lepidopterous pests in rice. A field experiment was conducted to evaluate the efficacy of chlorantraniliprole 0.4% gr (Ferterra 0.4% gr) against rice leaf folder and yellow stem borer during rainy (kharif) seasons 2017–19. The study revealed that broadcast application of chlorantraniliprole 0.4% gr @40 g a.i./ha at tillering stage was found effective and best insecticidal treatment in reducing rice leaf folder and yellow stem borer infestation with no phytotoxicity symptoms and increased grain yield. Chlorantraniliprole 0.4% gr @40 g a.i./ha was also safer to the natural enemies of the rice ecosystem. Post-harvest residual analysis of chlorantraniliprole 0.4% gr was found below the limit of quantification, i.e. 0.01 ppm in rice grains, soil, paddy straw and husk at recommended and double of the recommended dose indicating its safety to human, animals and environment.

Comprehensive Overview of Diamide Derivatives Acting as Ryanodine Receptor Activators.

The world's hunger is continuously rising due to conflicts, climate change, pandemics (such as the recent COVID-19), and crop pests and diseases. It is widely accepted that zero hunger is impossible without using agrochemicals to control crop pests and diseases. Diamide insecticides are one of the widely used green insecticides developed in recent years and play important roles in controlling lepidopteran pests. Currently, eight diamine insecticides have been commercialized, which target the insect ryanodine receptors. This review summarizes the development and optimization processes of diamide derivatives acting as ryanodine receptor activators. The review also discusses pest resistance to diamide derivatives and possible solutions to overcome the limitations posed by the resistance. Thus, with reference to structural biology, this study provides an impetus for designing and developing diamide insecticides with improved insecticidal activities.

Residue behavior of cyantraniliprole and its ecological effects on Procambarus clarkii associated with the rice-crayfish integrated system.

Cyantraniliprole, a second-generation diamide insecticide, was recently introduced in China, where the rice-crayfish integrated system (RCIS) is practiced to control rice pest infestations. The aim of this study was to investigate the residue behavior of cyantraniliprole in RCIS and its potential ecological effects on nontarget Procambarus clarkii in order to recommend safe pesticide application strategies. Cyantraniliprole dissipated in rice plants according to first-order kinetics, with an average half-life of 5.25 days and a dissipation rate of >95% over 28 days. The terminal cyantraniliprole residue levels in rice straw, paddy hull and brown rice were all within 0.2mg kg-1 , which is the China-recommended maximum residue limit. The tissues of P.clarkii accumulated and distributed cyantraniliprole in the descending order gill > hepatopancreas > intestine > muscles. Procambarus clarkii exposed to cyantraniliprole exhibited a moderate decrease in weight gain, specific growth rate and condition factor compared to the control group. Exposure of P.clarkii to cyantraniliprole caused histopathological alterations to the hepatopancreas, but the alterations were not statistically significant in the 60 g ai ha-1 cyantraniliprole group when compared with the control group. We suggest that 10% cyantraniliprole oil dispersion be sprayed twice at an interval of 14 days and dosage of 60 g ai ha-1 during the growth stage of rice in RCIS. © 2023 Society of Chemical Industry.

Anthranilic Diamides Containing Monofluoroalkene Amide Linkers as Potential Insect RyR Activators: Design, Synthesis, Bio-evaluation, and Computational Study.

In order to develop anthranilic diamides with novel chemotypes, a series of anthranilic diamides with acrylamide linkers were designed and synthesized. The results of preliminary bioassays indicated that compounds with a monofluoroalkene amide linker (Z-isomer) exhibited good larvicidal activity against lepidopteran pests. The LC50 values of compound A23 against Mythimna separata and Plutella xylostella were 1.44 and 3.48 mg·L-1, respectively, while those of chlorantraniliprole were 0.08 and 0.06 mg·L-1, respectively. Compound A23 also exhibited the same level of lethal potency against resistant and susceptible strains of Spodoptera frugiperda at 50 mg·L-1. Compound A23 exhibited similar symptoms as chlorantraniliprole in test larvae. Comparative molecular field analysis was conducted to demonstrate the structure-activity relationship. Central neuron calcium imaging experiments indicated that monofluoroalkene compounds were potential ryanodine receptor (RyR) activators and activated calcium channels in both the endoplasmic reticulum and the cell membrane. Molecular docking suggested that A23 had a better binding potency to P. xylostella RyR than chlorantraniliprole. The MM|GBSA dG bind value of A23 with P. xylostella RyR was 117.611 kcal·mol-1. Monofluoroalkene was introduced into anthranilic diamide insecticides for the first time and brought a novel chemotype for insect RyR activators. The feasibility of fluoroalkenes as insecticide fragments was explored.

Mechanisms underlying the effects of low concentrations of chlorantraniliprole on development and reproduction of the fall armyworm, Spodoptera frugiperda

It is well known that sublethal dose of insecticides induces life history trait changes of both target and non-target insect species, however, the underlying mechanisms remain not well understood. In this study, the effects of low concentrations of the anthranilic diamide insecticide chlorantraniliprole on the development and reproduction of the fall armyworm (FAW), Spodoptera frugiperda, were evaluated, and the underlying mechanisms were explored. The results showed that exposure of FAW to LC10 and LC30 chlorantraniliprole prolonged the larvae duration, decreased the mean weight of the larvae and pupae, and lowered the pupation rate as well as emergence rate. The fecundity of female adults was also negatively affected by treatment with low concentrations of chlorantraniliprole. Consistently, we found that exposure of FAW to LC30 chlorantraniliprole downregulated the mRNA expression of juvenile hormone (JH) esterase (SfJHE), leading to the increase of JH titer in larvae. We also found that treatment with low concentrations of chlorantraniliprole suppressed the expression of ribosomal protein S6 kinase1 (SfS6K1) in female adults, resulting in the downregulation of the gene encoding vitellogenin (SfVg). These results provided insights into the mechanisms underlying the effects of low concentrations of insecticides on insect pests, and had applied implications for the control of FAW.

Effect of different exposure times and doses of cyantraniliprole on oxidative stress and genotoxicity in earthworms (Eisenia fetida)

Cyantraniliprole, the second generation of diamide insecticides, is widely used to control various pests, which will certainly result in adverse effects on earthworms in soil. In this study, after exposure with six doses of cyantraniliprole (0, 0.5, 1, 2.5, 5, and 10 mg kg−1) by artificial soil method, six biomarkers, four functional genes, and histopathological changes of Eisenia fetida were measured on the 7th, 14th, 21st, and 28th days. The comprehensive toxicity was assessed by the IBR version 2 (IBRv2) method. The results showed that the reactive oxygen species (ROS) level was induced significantly. The superoxide dismutase (SOD) activity was activated in 7–28 days. The catalase (CAT) and glutathione S-transferases (GST) activities were also activated in the initial 14 days. The 8-hydroxy-2′-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) contents in the high treatment increased until the late stage of exposure. On the 28th day, the metallothionein (MT) and calreticulin (CRT) genes were up-regulated, the transcriptionally controlled tumor protein (TCTP) gene was down-regulated. The SOD gene showed a good correlation with SOD activity. Extensive histopathological damage was found in the endoderm and ectoderm of E. fetida. The 5 and 10 mg kg−1 treatments showed higher comprehensive toxicity than the 0.5, 1, and 2.5 mg kg−1 treatments on the 28th day. These results suggest that cyantraniliprole exerted certain subchronic toxic effects of oxidative stress, DNA damage, and histopathological changes to E. fetida, which provided theoretical basis for rational use of cyantraniliprole and evaluation of its safety to soil environment.