Plant immunity against herbivorous insects is often triggered by insect-derived salivary elicitors, yet the molecular mechanisms underlying their perception and signaling remain limited understood. Here, we identified a protein disulfide isomerase, MsPDI3, from the salivary glands of the oriental armyworm (Mythimna separata), which functions as an elicitor of plant defense. Transient expression of MsPDI3 in tobacco induced hypersensitive response-like necrosis and elevated salicylic acid (SA) levels. Mechanistically, MsPDI3 is secreted into maize plant through feeding, silencing MsPDI3 in larvae attenuated maize defense and enhanced larval growth, indicating its role in activating plant immunity. Using yeast two-hybrid screening and bimolecular fluorescence complementation, we demonstrated that MsPDI3 interacts with a maize Src homology 3 domain-containing protein (ZmSH3), at the plasma membrane. Virus-induced gene silencing of ZmSH3 abolished MsPDI3-triggered SA accumulation and compromised plant resistance, suggesting ZmSH3 as a key component in the defense signaling cascade. These findings established a novel link between SH3-mediated signal transduction in mediating insect-derived elicitor perception and SA-dependent defense activation, providing new insights into the molecular dialogue between plants and chewing insects.

Terpinolene, a monoterpene volatile, has been demonstrated to exhibit repellent activity against Bemisia tabaci adults. However, molecular basis of B. tabaci identifying and responding to terpinolene was unclear. Chemosensory proteins (CSPs) family is one of the most important member in insect olfaction system which plays a crucial role in hosts selection. Here, we cloned 13 BtabCSP genes in B. tabaci MED and detected transcriptional response to terpinolene. Based on proteins structure prediction, molecular docking, protein expression and fluorescence competition binding assay, BtabCSP1and BtabCSP12 exhibited strong binding affinity to terpinolene. Moreover, RNA interference (RNAi) combined with Y-tube assays showed that silencing BtabCSP1 or BtabCSP12 genes weakened the repellency effect of terpinolene to B. tabaci. Our results not only revealed molecular mechanism of CSPs-mediated repellency of terpinolene against B. tabaci, but also provided a theoretical basis for terpinolene-based repellent for pest management.

Using gas chromatography-electroantennographic detection (GC-EAD), gas chromatography-mass spectrometry (GC-MS), and chiral capillary GC with synthetic stereoisomers, followed by laboratory and field validation, we identified the sex pheromone of the copper green chafer (Anomala corpulenta) as (R)-japonilure, with its antipode, (S)-japonilure, acting as a behavioral antagonist. Single sensillum recordings (SSR) revealed two types of pheromone-responsive sensilla, one narrowly tuned to the sex pheromone, and another that responded to both (R)- and (S)-japonilure, albeit with lower sensitivity to the latter. Functional characterization of A. corpulenta odorant receptors (ORs) by two-electrode voltage clamp (TEVC) led to the identification of two key receptors, AcorOR18, which was selective for (R)-japonilure, and AcorOR29, which was activated by both enantiomers, and 10 ORs sensitive to plant-derived compounds. SSR combined with fluorescence in situ hybridization (FISH) experiments suggested that AcorOR18 is expressed in pheromone-specific sensilla, whereas AcorOR29 is localized in sensilla responsive to both the pheromone and its antagonist. RNA interference (RNAi) knockdown of either receptor reduced male attraction to (R)-japonilure, while simultaneous knockdown of both genes abolished behavioral responses, confirming that both receptors are required for full pheromone detection. Three orthologous receptors were identified in the Japanese beetle (Popillia japonica) genome. Two of them, PjapOR42 and PjapOR37, exhibited strict enantioselectivity for (R)-japonilure, whereas PjapOR39 showed a relaxed enantiospecificity, responding to both enantiomers and preferentially to the behavioral antagonist. These results suggest that PjapOR39 may mediate detection of (S)-japonilure and contribute to behavioral antagonism, revealing a shared molecular mechanism of enantiomeric discrimination in scarab beetles.

The tomato pinworm, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a globally invasive and highly destructive pest that primarily targets solanaceous plants. This species has developed resistance to most currently used insecticides. Broflanilide, a novel meta-diamide insecticide that binds to a unique site on the gamma-aminobutyric acid receptor, has been effectively used for managing Lepidopteran pest species. This study aimed to explore the development of resistance, fitness effects, and underlying biochemical mechanisms. A laboratory strain of T. absoluta was exposed to broflanilide for 17 generations, showing no apparent resistance development. The low realized heritability of 0.0804 indicated a minimal risk of resistance. Fifteen field-collected populations of T. absoluta were surveyed and exhibited high susceptibility to broflanilide, with resistance ratios ranging from 0.63 to 3.53. The activities of major detoxification enzymes (cytochrome P450 monooxygenase, carboxylesterase, and glutathione S-transferase) remained unchanged after broflanilide selection. Life table analysis revealed extended larval stages and reduced adult longevity, fecundity, and oviposition duration in the broflanilide-selected strain, but without significantly affecting overall fitness. These findings provide valuable insights for formulating practical guidelines for the effective use of broflanilide in managing insecticide resistance in T. absoluta.

Bemisia tabaci is a major agricultural pest, and its widespread resistance to multiple insecticides poses a significant challenge for effective control. Dimpropyridaz, a new pyridazine-pyrazolecarboxamide insecticide, demonstrates high toxicity against sap-sucking pests, including those resistant to other class insecticides. However, the mechanisms underlying B. tabaci detoxification of dimpropyridaz remain unclear. In this study, transcriptome sequencing was performed on B. tabaci adults exposed to a lethal dose of dimpropyridaz for 48 h, identifying 973 differentially expressed genes (DEGs). GO and KEGG enrichment analyses revealed significant involvement of cytochrome P450 monooxygenases (P450s) in the detoxification pathway. Among the DEGs, a novel P450 gene, CYP402C18, was upregulated by 4.05-fold in response to dimpropyridaz and classified within the Clan3 CYP6 subfamily. Expression profiling revealed that CYP402C18 expression peaked in adults, with thoracic and head tissues exhibiting 5.18- and 1.64-fold higher levels, respectively, compared to the abdomen. Silencing CYP402C18 via dsRNA feeding significantly increased B. tabaci mortality to dimpropyridaz by 11.8 %. Molecular docking and dynamic simulations indicated stable binding between dimpropyridaz and CYP402C18, with a binding free energy of -35.89 kcal/mol. Notably, the alanine scanning results demonstrated that mutating the residues of Phe502 and Thr116 to alanine in CYP402C18 significantly reduced the binding energy with dimpropyridaz, indicating that these amino acids play a crucial role in the interaction with dimpropyridaz. These findings demonstrate that CYP402C18 contributes to dimpropyridaz detoxification and may be involved in resistance development, providing a theoretical basis for more targeted and sustainable use of dimpropyridaz in field management.

The tomato leafminer (Tuta absoluta), a globally invasive pest, poses a major economic threat to tomato production. Although chemical control remains the primary management method, sustainable alternatives are urgently needed. Sex pheromone communication is critical for moth courtship and mating, with pheromone-binding proteins (PBPs) playing a key role in this process. In this study, we identified a PBP gene, TabsPBP2, from the T. absoluta transcriptome. Real-time quantitative PCR (RT-qPCR) revealed that TabsPBP2 is highly expressed in the antennae, with a strong male-biased expression pattern. Ligand-binding assays demonstrated that TabsPBP2 has the highest affinity for the sex pheromones (3E, 8Z, 11Z)-tetradecatrienyl acetate (TDTA) and (3E, 8Z)-tetradecadienyl acetate (TDDA). It also demonstrated a moderate-to-strong binding affinity to several tomato volatiles, including 2-carene, myrcene, α-pinene, cis-3-hexen-l-ol, methyl salicylate, sabinene, and α-terpinene. Molecular docking suggested that hydrophobic interactions predominantly stabilize the TabsPBP2–ligand complexes, with PHE118, PHE12, LEU90, LEU68, and ALA73 identified as key interacting residues. Electroantennogram (EAG) and Y-tube olfactometer assays confirmed that TDTA and TDDA act as strong attractants for male T. absoluta. This study enhances our understanding of the pheromone recognition in T. absoluta and provides a foundation for developing novel, pheromone-based pest control strategies.

Spodoptera litura is a globally important agricultural pest whose increasing resistance to insecticides threatens effective control. Tetraniliprole, a novel diamide insecticide targeting the ryanodine receptor, was developed recently, yet high levels of resistance have already emerged. Understanding the mechanisms of resistance and any other associated fitness costs is essential for resistance management. In this study, we established a near-isogenic line (NIL) of S. litura through backcrossing a laboratory-susceptible strain and a field-resistant population. The NIL strain showed 157.4-fold resistance to tetraniliprole and 32.7-fold resistance to chlorantraniliprole. Mutational analysis confirmed the absence of the I4723 M/K mutations commonly linked to diamide resistance. Synergist and enzyme activity assays revealed that piperonyl butoxide (PBO) produced synergistic ratios of 1.48 and 1.42 against tetraniliprole and chlorantraniliprole, respectively. Additionally, cytochrome P450 activity in the NIL strain was 1.6 times higher than in the susceptible strain, suggesting a role of P450-mediated metabolism in resistance to both compounds. Life table analysis further demonstrated that compared with the susceptible strain, the NIL strain exhibited a mean generation time shortened by 1.16 days, a net reproductive rate increased by 1.14-fold, and a relative fitness of 1.13, indicating no fitness cost associated with tetraniliprole resistance. These findings highlight the rapid development of tetraniliprole resistance in S. litura driven by enhanced P450 activity and the absence of detectable fitness penalties. The results provide a theoretical basis for the rational rotation of tetraniliprole with other insecticides and support resistance management strategies for S. litura.

Lycium barb arum,known as wolfberry or goji berry,is consumed by humans as a medicine and a food homology product.Conventionally grown wolfberry is often treated extensively with pesticides,which could pose a hazard to humans.Here,the degradation dynamics of dinotefuran and its 2 metabolites(1-methyl-3-（tetrahydro-3-furylmethyl） urea（UF） and 1-methyl-3-（tetrahydro-3-furylmethyl） guanidine（DN）),during wolfberry cultivation and processing was investigated.The half-life(T 1/2 ) of dinotefuran was 11.36 and 9.76 days,respectively,under the recommended dosage and double the recommended dosage.During the oven and sun drying processes,processing factors（PFs） of dinotefuran were 1.07-1.34,implying the enrichment of pesticide residues.Decoction process made the removal rate of dinotefuran reach 87.48%,which is higher than that of the brewing process（14.7%）,while dinotefuran remained in the wine with high ethanol content in the alcohol soaking process.The hazard quotient（HQ） of dinotefuran,as determined via dietary risk assessment combined with PFs,was <1,indicating an acceptable risk for human consumption.Bioaccessibility of dinotefuran in the three digestive stages were intestinal（18.20%-88.08%）> gastric（5.45%-86.72%）> oral（23.18%） via in vitro simulated digestive system.These findings provide scientific evidence for reasonable application and risk assessment of dinotefuran residues in wolfberry.

Huangjiu, a traditional Chinese rice wine, faces safety issues from contaminants like pesticide residues, mycotoxins, toxic ions, and sediments. These contaminants not only affect Huangjiu quality but also threaten consumers health. Pesticide residues and mycotoxins primarily originate from raw materials, whereas toxic ions, including heavy metals, mainly result from environmental pollution. Effective control of these contaminants requires enhanced agricultural practices, stricter raw materials regulation, and optimized processing techniques. Precipitation in Huangjiu, caused by protein-polyphenol complexes and oxidation, can be mitigated by optimizing the storage conditions. Despite advances in detection and control methodologies, however, challenges remain due to the lack of standardized contaminant limits. Future research should focus on developing more sensitive detection techniques, enhancing control strategies, and evaluating long-term health effects of these contaminants to ensure the sustained safety and quality of Huangjiu.

Spodoptera litura, a globally destructive agricultural pest, has evolved significant resistance to many popular insecticides. Broflanilide, a new meta-diamide pesticide that targets the specific site of γ-aminobutyric acid (GABA) receptor, provides broad-spectrum control against pests in the orders Lepidoptera and Thysanoptera. However, its potential resistance risk in S. litura remains unclear. In this study, a laboratory-selected strain was generated through ten consecutive generations of selection with broflanilide, revealing a low overall risk for resistance development. Concurrently, field resistance monitoring across China indicated that most S. litura populations remained highly susceptible, while the Sanya (SY) population exhibited moderate resistance (26.8-fold). Following seven additional generations of selection, the SY population evolved high resistance (56.2-fold) and was designated the SY-R strain. Using SY-R, we investigated inheritance, cross-resistance, and mechanism of detoxification underlying field-evolved resistance. Broflanilide showed minimal cross-resistance to the tested pesticides. Genetic and synergistic analyses indicated that resistance was incompletely dominant, autosomal, and polygenic, primarily associated with detoxification of glutathione S-transferase (GST) and cytochrome P450 (P450). These findings provide a comprehensive assessment of broflanilide resistance risk and provide valuable information and data for developing effective strategies of pesticide resistance management.

The pharmacological effects of Ophiopogon japonicus are influenced by its active substances like saponins and flavonoids, which are crucial for assessing its quality. An ultra-high performance liquid chromatography tandem triple quadrupole mass spectrometry (UHPLC-MS/MS) method was developed for the qualitative and quantitative assessment of 20 saponins and flavonoids in O. japonicus. The average recovery of these compounds ranged from 70 % to 130 %, with relative standard deviations (RSD) between 0.11 % and 16.46 %. Planting O. japonicus in Santai County (Sichuan, China) and investigated the impact of 0.6, 1.3, 2.8, 5.6, and 11.3 g/L concentrations of paclobutrazol on both root yield and the levels of saponins and flavonoids in O. japonicus. Lower paclobutrazol concentrations (1.3 g/L) led to increased levels of saponins and flavonoids in O. japonicus such as ophiopogonin D ', ruscogenin and ophiopogonanone F, with paclobutrazol residues measuring 77.17 mu g/kg DW. This preliminary insight into the influence of paclobutrazol and production of saponin flavonoids in O. japonicus bears significance for its larger industrial production.

Invasive pest species of economic importance often require intensive pesticide use, leading to recurrent problems of pesticide resistance. Such is the case with the tomato pinworm, Phthorimaea (=Tuta) absoluta (Meyrick) (Lepidoptera: Gelechiidae), requiring the use of new insecticidal compounds for their management. Tetraniliprole, a novel insecticide effective against lepidopteran, coleopteran, and dipteran pests, is a potential alternative against the tomato pinworm. This study investigates the sublethal effects of tetraniliprole on the development and reproduction of the parental (F0) and progeny (F1) generations of P. absoluta while exploring the underlying mechanisms. Sublethal tetraniliprole exposure extended larval duration and reduced pupation rate, emergence rate, hatchability, and fecundity in both the F0 and F1 of P. absoluta. Additionally, the life-table parameters of the F1 generation were significantly altered, with decreases in net reproductive rate (R0) and increases in the mean generation time (T), leading to decreases in the intrinsic rate of population growth (r). The relative fitness of F1 insects exposed to tetraniloprole was reduced compared to unexposed insects (0.63 and 0.49, respectively). Furthermore, the mRNA expression levels of reproduction-related genes such as TaVg and TaVgR were down-regulated in the tetraniliprole-exposed insects. Finally, insect exposure to tetraniloprole increases the activity of the antioxidant enzymes such as glutathione S-transferase (GSTs), catalase (CAT), and superoxide dismutase (SOD). Taken together, these findings suggest that sublethal concentrations of tetraniloprole adversely affect both the development and reproduction of P. absoluta, providing a foundation for optimizing pest control strategies.

Bemisia tabaci is a formidable insect pest worldwide, and exhibits significant resistance to various insecticides. Flupyradifurone is one novel butenolide insecticide and has emerged as a new weapon against B. tabaci , but fieldevolved resistance to this insecticide has become a widespread concern. To unravel the mechanisms of fieldevolved flupyradifurone resistance, we conducted a comprehensive investigation into susceptibility of twentyone field populations within the Beijing -Tianjin -Hebei Region of China. Alarmingly, thirteen of these populations displayed varying degrees of resistance, ranging from low to medium levels, and building upon our prior findings, we meticulously cloned and characterized the CYP6CX4 gene in B. tabaci . Our investigations unequivocally confirmed the association between CYP6CX4 overexpression and flupyradifurone resistance in three of the thirteen resistant strains via RNA interference. To further validate our findings, we introduced CYP6CX4 overexpression into a transgenic Drosophila melanogaster line, resulting in a significant development of resistance to flupyradifurone in D. melanogaster . Additionally, homology modeling and molecular docking analyses showed the stable binding of flupyradifurone to CYP6CX4 , with binding free energy of -6.72 kcal mol - 1 . Collectively, our findings indicate that the induction of CYP6CX4 exerts one important role in detoxification of flupyradifurone, thereby promoting development of resistance in B. tabaci.

Insecticides have been instrumental in the prevention and management of key agricultural insect pests, thereby contributing to increased food production. However, the effectiveness of insecticides diminishes when targeted pests develop resistance, a phenomenon linked to species evolution and survival instincts. The Arthropod Pesticide Resistance Database, maintained by Michigan State University, has documented 18,934 cases of resistance to insecticidal active ingredients worldwide from 1914 to the present. This special collection comprises eight original research articles that delve into the monitoring, mechanism and management of insecticide resistance. Two papers present multi-year resistance monitoring results in Chilo suppressalis and Spodoptera litura. Two studies employ transcriptome data to examine the candidate genes implicated in solanine-induced response in Phthorimaea operculella and chlorantraniliprole-induced response in Galeruca daurica, respectively. Additionally, two papers describe the genetic and physiological impacts of insecticides, while others explore the potential mechanisms underlying chlorfenapyr resistance in Glyphodes pyloalis and pyrethroids resistance in Laodelphax striatellus. The new results, conclusions and suggestions presented in this collection are anticipated to contribute to the advancement of knowledge on insecticide resistance and facilitate the development of a sustainable approach to managing insect pests.

Bemisia tabaci is one of the most destructive agricultural insect pests around the world, and it has developed high levels of resistance to most pesticides. Dimpropyridaz, a novel insecticide developed by BASF, displays excellent activity against piercing-sucking insect pests. In this study, baseline of susceptibility showed all tested field populations of B. tabaci are susceptible to dimpropyridaz. After continuous selection with dimpropyridaz in the lab, a B. tabaci strain (F12) developed 2.2-fold higher level of resistance compared with a susceptible MED-S strain, and the realized heritability (h2) was estimated as 0.0518. The F12 strain displayed little cross-resistance to afidopyropen, cyantraniliprole, sulfoxaflor, or abamectin, and significantly increased activity of cytochrome P450 monooxygenase (P450). The fitness cost of dimpropyridaz resistance was evident in F12 strain, which had a relative fitness of 0.95 and significantly lower fecundity per female compared with MED-S strain. Taken together, B. tabaci displays high susceptibility to dimpropyridaz in the field, and low risk of developing resistance to dimpropyridaz under successive selection pressure. Little cross-resistance to popular insecticides was found, and fitness cost associated dimpropyridaz resistance was observed. Higher activity of cytochrome P450 in the F12 strain, may be involved in the process of detoxifying dimpropyridaz in whitefly.

Spodoptera 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 F-1 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.

Bemisia tabaci is a formidable insect pest worldwide, and it exhibits significant resistance to various insecticides. Dimpropyridaz is a novel pyridazine pyrazolecarboxamide insecticide used against sucking insect pests, but there is little information regarding its metabolic detoxification in arthropods or cross-resistance with other insecticides. In this study, we found that dimpropyridaz shows no cross-resistance with three other popular insecticides, namely abamectin, cyantraniliprole, and flupyradifurone. After treatment of B. tabaci adults with a high dose of dimpropyridaz, higher cytochrome P450 monooxygenase (P450) activity was detected in the survivors, and the expression of the P450 gene CYP6DW4 was highly induced. Cloning and characterization of the full-length amino acid sequence of CYP6DW4 indicated that it contains conserved domains typical of P450 genes, phylogenetic analysis revealed that it was closely related to a B. tabaci protein, CYP6DW3, known to be involved in detoxification of imidacloprid. Silencing of CYP6DW4 by feeding insects with dsRNA significantly increased the susceptibility of B. tabaci to dimpropyridaz. In addition, homology modeling and molecular docking analyses showed the stable binding of dimpropyridaz to CYP6DW4, with binding free energy of -6.65 kcal/mol. Our findings indicate that CYP6DW4 plays an important role in detoxification of dimpropyridaz and possibly promotes development of resistance in B. tabaci.

Tuta absoluta is one of the most devastating and invasive insect pests throughout the world. It feeds on numerous solanaceous plant species and has evolved resistance to various types of popular chemical agents. Tetraniliprole is a novel diamide chemical agent that acts as a modulator of the ryanodine receptor, and it has been considered as a promising insecticide against T. absoluta. In this study, we subjected a T. absoluta strain to 17 generations of continuous selection with tetraniliprole in the lab. The resulting tetraniliprole-selected T. absoluta strain (YN-Tet) exhibited a 12.9-fold higher resistance to tetraniliprole compared with a susceptible strain (YN-S), and the realized heritability (h2) was estimated to be 0.1437. The YN-Tet strain exhibited a low level of cross-resistance to chlorantraniliprole and significantly higher cytochrome P450 monooxygenase activity. There was a fitness cost of tetraniliprole resistance in the YN-Tet strain, which had a relative fitness of 0.83 compared with the YN-S strain and a significantly lower fecundity per female. These results enhance our knowledge of the risk and mechanism of tetraniliprole resistance, enabling future optimization of resistance management strategies.

The tomato pinworm, Phthorimaea (=Tuta) absoluta, is considered one of the most destructive and invasive insect pests worldwide, having developed significant resistance to many popular insecticides. In this study, we monitored the field resistance of P. absoluta populations from China to three diamide insecticides: flubendiamide, chlorantraniliprole, and cyantraniliprole. We found that one field population from Wuzhong City (WZ) exhibited high level of resistance to chlorantraniliprole. Using the WZ population and a susceptible reference strain (YN-S), we established a near-isogenic line (WZ-NIL) of P. absoluta with resistance to chlorantraniliprole. This strain also showed substantial cross-resistance to flubendiamide, and cyantraniliprole. Genetic analysis revealed that the inheritance of resistance to chlorantraniliprole in the WZ-NIL strain was autosomal and incompletely dominant. Additionally, the pesticide synergist piperonyl butoxide significantly inhibited chlorantraniliprole resistance by compromising P450 monooxygenase activity, which was significantly higher in the resistant strain. Furthermore, WZ-NIL had significantly prolonged developmental stages, lower pupation rates, reduced female fecundity, and lower egg hatchability than YN-S individuals. The fitness of WZ-NIL relative to YN-S was estimated to be 0.73, indicating significant fitness cost associated with chlorantraniliprole resistance. Rotating chlorantraniliprole with other insecticides that have different modes of action and degradation may be particularly useful for managing chlorantraniliprole resistance in P. absoluta.