Porous hollow silica microcapsules enable stable, sustainable pesticide formulations, but their synthesis is complex and solvent-dependent. In this study, a green method was developed using a choline-based ionic liquid as both a solvent and soft template. The resulting choline-chlorantraniliprole-porous hollow silica (CHO-CHL-PHS) microcapsules achieved a chlorantraniliprole loading of 47.17%, exhibited a spherical morphology, and were synthesized without organic solvents or additional surfactants. The microcapsules improved the photostability of chlorantraniliprole, extending its UV half-life from 0.89 to 1.27 h, and showed sustained diffusion release at pH 5.0-10.0. Laboratory bioassays demonstrated that CHO-CHL-PHS possessed significantly higher virulence against Locusta migratoria manilensis nymphs than technical-grade CHL. Persistence trials confirmed that its efficacy within 7 days was superior to that of a commercial suspension concentrate. Safety assessments showed no phytotoxicity on ryegrass or corn and reduced toxicity to zebrafish. This study offers a promising strategy for developing effective and environmentally compatible agrochemical formulations.

Apoptosis is regulated by a conserved gene network across species for maintaining homeostasis and stress response. While intracellular apoptotic pathways are well characterized, the extracellular mechanisms governing apoptosis remain largely unexplored, probably due to divergence in species-specific gene sets involved in extracellular regulation. Here, we uncover a novel extracellular apoptotic mechanism mediated by acetylcholine-binding protein 5 (AChBP5) in the wolf spider Pardosa pseudoannulata. AChBP5 is highly expressed in the spider fat-body (midgut diverticula), while other four AChBP genes are abundantly expressed in the tissue brain. Among five AChBP genes, only AChBP5 exhibited broad transcriptional induction upon exposure to various insecticides, including neonicotinoids and other classes. AChBP5 expression was also upregulated by reactive oxygen species (ROS) including H2O2, key triggers of apoptosis. Functional assays demonstrated that, in cultured cells, AChBP5 acted as an extracellular sacrifice buffer against oxidative stress to maintain cell viability, becoming progressively inactivated by ROS in a concentration-dependent manner. RNAi-mediated silencing of AChBP5 significantly increased spider susceptibility to both-induced and direct oxidative stress, underscoring its critical protective function. Collectively, these findings support a model in which lineage-specific genes, AChBP5 may contribute to extracellular modulation of apoptosis and provide a mechanism by which spiders could respond to chemical stressors.

BACKGROUND:In recent years, Monolepta hieroglyphica has become a destructive pest threatening food security in northeastern China, where M. hieroglyphica adults inflict substantial yield losses through feeding on young leaves, silks and sometimes on developing kernels from maize. Plant-mediated RNA interference (RNAi) has proven to be a promising tool for sustainable pest management. Thus, specific and effective RNAi target genes for M. hieroglyphica management are needed. RESULTS:Using homology-based searching of the M. hieroglyphica genome, we identified two chitin synthase genes, and named them MhCHS1 and MhCHS2, respectively. MhCHS2 was found to be expressed in the midgut of M. hieroglyphica. We found that after feeding on transgenic maize expressing MhCHS2 double-stranded RNA (dsRNA) for 2 days, the expression of MhCHS2 was significantly reduced, and the formation of peritrophic membrane was inhibited in M. hieroglyphica. Notably, after feeding on maize expressing MhCHS2 dsRNA, half of M. hieroglyphica died within 4 days, with the majority succumbing by 7 days. Maize expressing MhCHS2 dsRNA exhibited a high degree of resistance to M. hieroglyphica. CONCLUSION:These findings further validate the essential role of MhCHS2 in maintaining formation of the peritrophic membrane and ensuring organism survival in M. hieroglyphica. This study suggests that MhCHS2 might be an ideal RNAi target for pest management, and maize expressing MhCHS2 dsRNA could be a potential and effective strategy for M. hieroglyphica control. © 2026 Society of Chemical Industry.

Background Galeruca daurica (Joannis), a notorious pest in Northern China, seriously threatens Inner Mongolia grassland ecosystems and economy, due to its strong cold hardiness and huge outbreak in a short time. Despite previous studies emphasized molecular mechanisms in response to low-temperature stress, especially based on studies of key gene functions on cold hardiness, there is a lack of in-depth research on transcriptional regulation mechanisms. Results The G . daurica eggs, collected from the grassland of Xianghuang Banner of Xilin Gol League in Inner Mongolia, were reared to the 1st and 2nd instar larvae at different temperatures. RNA-Seq analyzed differentially expressed genes (DEGs) in larvae reared in different states. Weighted gene co-expression network analysis (WGCNA) identified cold hardiness-related gene modules, and regulatory network analyses screened key transcription factors (TFs). The Gene Transcription Regulation Database (GTRD) predicted HSF1 target genes. The genes of HSF1 , HSP26 , and HSP68 were knocked down to evaluate the impacts on expression and cold hardiness. The blue module showed the highest cold hardiness correlation, with HSF1 as a key regulator. Silencing HSF1 decreased HSP26 and HSP68 expression, and knocking down these HSPs reduced the expression of HSF1 . Silencing HSF1 , HSP28 , and HSP86 can increase super-cooling points (SCPs) and freezing points (FPs) significantly. HSF1 regulates HSP26 and HSP68 expression to enhance the cold hardiness of G. daurica . These results elucidate the molecular mechanisms underlying cold hardiness and provide new insights into the adaptability of insects to extreme environments. Conclusions The RNA-Seq analysis of G. daurica has revealed a potential regulatory relationship among HSF1 , HSP26 , and HSP68 . The increased SCPs and FPs after RNAi support their roles in low-temperature tolerance. These findings provide preliminary insights into the molecular basis of cold hardiness in G. daurica.

RNA interference (RNAi) represents a promising pest control strategy, applicable to both insect-resistant genetically modified (IRGM) crops and sprayable RNAi insecticides. These products can achieve sequence-specific gene silencing and require rigorous environmental risk assessment (ERA) prior to approval. However, current environmental safety assessments of RNAi products and other RNAi experiments frequently use double-stranded EGFP (dsEGFP) as a negative control, while suitable RNAi-based positive controls are lacking. Sometimes conventional chemical toxins (e.g., chlorpyrifos) or protein inhibitors (e.g., trypsin inhibitors) are used as substitutes, but their distinct mechanisms, persistence, and metabolism make them inappropriate for RNAi-specific evaluations. In this study, we evaluated the suitability of RNAi-based positive controls for assessing non-target effects on Harmonia axyridis, a widely distributed predatory beetle used as a bioindicator in biosafety assessments. Under laboratory conditions, we tested one microRNA (miR-92a) and two double-stranded RNAs (dsHaSnf7 and dsHaDiap1) for their effects on H. axyridis. Injection of miR-92a showed no significant difference in mortality compared to controls, whereas dsHaSnf7 and dsHaDiap1 significantly reduced survival rates and target gene expression, as confirmed by qPCR. These findings suggest that HaSnf7 and HaDiap1 are suitable candidate genes for establishing RNAi-specific positive controls in environmental risk assessments of RNAi-based products.

Hermonassa cecilia is a Lepidoptera pest primarily distributed in East Asia, belonging to the subfamily Noctuinae, which includes species that typically target and damage the underground parts of plants. However, there is limited information available on the life history and genomic resources of H. cecilia to date. In this study, we present a high-quality reference genome of H. cecilia generated using PacBio sequencing and Hi-C methods. The assembled genome size is 626.10 Mb, with a N50 of 21.00 Mb, and the contigs were mapped onto 31 chromosomes. BUSCO analysis indicated high genome completeness, with a score of 99.40%. We identified 281.45 Mb of repetitive sequences, which account for 44.21% of the genome, and annotated 22,662 protein-coding genes, 89.29% of which had functional annotations. This study represents the first assembly and annotation of the H. cecilia genome, providing a valuable resource for understanding its biological characteristics and offering significant potential for comparative genomics within the Noctuinae subfamily.

Livestock grazing is a predominant land use in grasslands globally, primarily for economic purposes. It can also be a management practice to maintain ecosystem health when implemented sustainably, but overgrazing can lead to degradation. Grasshoppers are integral to grassland ecosystems and can indicate ecosystem functioning and services. Understanding insect responses to human activities, particularly land-use changes, is crucial for ecology and conservation as it impacts biodiversity. This study analysed data from a grazing experiment in Inner Mongolia, China, to examine the effects of grazing intensity on grasshopper communities. Total grasshopper abundance increased with grazing intensity, while Calliptamus abbreviatus abundance decreased. Overall grasshopper diversity declined with increasing grazing intensity. Dasyhippus barbipes abundance was regulated by plant diversity, Oedaleus asiaticus by the above-ground nitrogen pool of dominant plants, Myrmeleotettix palpalis by perennial grass biomass and C. abbreviatus by shrub and semi-shrub biomass. Grasshopper diversity was significantly affected by perennial grass biomass. Light grazing intensity can maintain grasshopper diversity while keeping abundance low, providing a basis for effective grazing management strategies in desert steppe.

Mosquitoes are important vectors of human diseases, with Anopheles sinensis posing substantial health risks across Asia. Although pyrethroids remain widely deployed, their effectiveness is compromised by resistance associated with sodium channel mutations. Spider peptide neurotoxins are potential biopesticides. Here, we evaluated the insecticidal efficacy of the Pardosa pseudoannulata neurotoxin PPTX-04, both alone and in combination with pyrethroids, against A. sinensis. Bioassays were conducted on laboratory and field populations, and functional analyses were performed on wild-type and mutant sodium channels (AsNav1-1). The co-toxicity of PPTX-04 with etofenprox was assessed across different mixture ratios. To enhance applicability, a nano-formulated version (Nano-PPTX-04) was developed and tested. PPTX-04 displayed potent activity against A. sinensis and retained efficacy against the L1014 sodium channel mutation that significantly reduced pyrethroid susceptibility. Mixtures of PPTX-04 and etofenprox produced synergistic toxicity at specific ratios (e.g., 3:1, 2:1). Notably, Nano-PPTX-04 exhibited superior potency relative to the native peptide, and its combinations with etofenprox yielded pronounced synergistic effects, particularly in pyrethroid-resistant field populations. Collectively, our laboratory-scale findings highlight the potential of PPTX-04, especially in nano-formulated form and in synergy with pyrethroids, as a potential strategy for mosquito control. This approach warrants further investigation as a viable solution to pyrethroid resistance.

The prolonged use of pyrethroid insecticides for controlling the plant bug Lygus pratensis has led to upward resistance. This study aims to elucidate the molecular mechanisms and potential regulatory pathways associated with lambda-cyhalothrin resistance in L. pratensis. In this study, we constructed a regulatory network by integrating transcriptome RNA-Seq and proteome iTRAQ sequencing analyses of one lambda-cyhalothrin-susceptible strain and two resistant strains, annotating key gene families associated with detoxification, identifying differentially expressed genes and proteins, screening for transcription factors involved in the regulation of detoxification metabolism, and examining the metabolic pathways involved in resistance. A total of 82,919 unigenes were generated following the assembly of transcriptome data. Of these, 24,859 unigenes received functional annotations, while 1064 differential proteins were functionally annotated, and 1499 transcription factors belonging to 64 distinct transcription factor families were identified. Notably, 66 transcription factors associated with the regulation of detoxification metabolism were classified within the zf-C2H2, Homeobox, THAP, MYB, bHLH, HTH, HMG, and bZIP families. Co-analysis revealed that the CYP6A13 gene was significantly up-regulated at both transcriptional and translational levels. The GO and KEGG enrichment analyses revealed that the co-up-regulated DEGs and DEPs were significantly enriched in pathways related to sphingolipid metabolism, Terpenoid backbone biosynthesis, ABC transporters, RNA transport, and peroxisome function, as well as other signaling pathways involved in detoxification metabolism. Conversely, the co-down-regulated DEGs and DEPs were primarily enriched in pathways associated with Oxidative phosphorylation, Fatty acid biosynthesis, Neuroactive ligand-receptor interactions, and other pathways pertinent to growth and development. The results revealed a series of physiological and biochemical adaptations exhibited by L. pratensis during the detoxification metabolism related to lambda-cyhalothrin resistance. This work provided a theoretical basis for further analysis of the molecular regulation mechanism underlying this resistance.

Exogenous methyl jasmonate is widely acknowledged for its role in triggering plants’ defense systems against pest invasions. Nonetheless, there has been a dearth of research exploring the elicitation of defense mechanisms by jasmonic acid in alfalfa. In order to investigate the effect of methyl jasmonate on thrips resistance in alfalfa, Medicago sativa L.cv. Caoyuan No. 4 was exogenously sprayed with different concentrations of methyl jasmonate, and thrips and Orius strigicollis (natural enemies) behavioral choice, physiological and transcriptomic analyses were performed. The results revealed a concentration-dependent inducible effect of methyl jasmonate on the behavioral choice, feeding and oviposition of thrips mediated by volatile organic compounds. Moreover, methyl jasmonate treatment at varying concentrations significantly influenced the activity levels of defense enzymes and secondary metabolites in alfalfa. Notably, the most pronounced induction effect of methyl jasmonate was observed at a concentration of 0.1 mmol/L, particularly evident in the enhanced activity of peroxidase, polyphenol oxidase, lipoxygenase and tannins. Transcriptome analysis showed that differentially expressed genes between methyl jasmonate treatment and control group (methyl jasmonate-free treatment) were mainly enriched in metabolic pathways and plant hormone signal transduction pathways such as terpenoid biosynthesis, linoleic acid metabolism and jasmonate signal transduction. Subsequent pathway analysis elucidated the potential of methyl jasmonate treatment to elevate endogenous jasmonic acid levels and instigate the activation of the jasmonate signaling pathway.

Orthopteran insects, which serve a wide range of biogeographic and ecological functions, play multiple roles in maintaining the health and stability of grassland ecosystems. Consequently, they are considered important indicators of ecological changes in grasslands. To assess and maintain ecosystem functionality, it is essential to understand the ecological niches of Orthopteran insects. This includes analyzing niche breadth, interspecific relationships, biological requirements, tolerances, and niche overlap among sympatric species. However, how these ecological dynamics respond to grazing in desert steppe ecosystems remains largely unexplored. Through a grazing experiment initiated in 2004 on desert steppes, with sampling conducted in 2021 and 2022, this study analyzed the response of grasshopper niche width, niche overlap, and interspecific connectivity under different grazing intensities, providing insights into the effects of long-term grazing on the dominant insect group. Results showed that the increase in grazing intensity significantly expanded the niche widths of Myrmeleotettix palpalis and Oedaleus decorus asiaticus, while significantly reducing the niche width of Calliptamus abbreviates. The overlap of grasshopper niches significantly decreased with increasing grazing intensities, suggesting a strategy to mitigate competition amidst reduced plant resources. Concurrently, interspecific associations shifted from non-significant to significantly positive, indicating a potential for ecological facilitation and increased species interdependence under higher grazing pressures. Increases in grazing intensities resulted in grasshoppers adapting their strategies for plant resource utilization in response to environmental changes, which helped to mitigate direct interspecific competition and may have contributed to the development of a more stable ecosystem state.

Megalurothrips usitatus (Bagnall) is one of the most important pests harming alfalfa (Medicago sativa L.) in Ningxia. In the field, Orius strigicollis (Poppius) is abundant. It has a high search efficiency and consumption rate of prey on a wide range of thrips and is often used as a biological control agent for these insects. To understand the predatory function and biological control potential of O. strigicollis on M. usitatus, the predatory ability of adults of O. strigicollis on second-instar larvae and adults of M. usitatus was measured under indoor conditions, and the Holling II and Hassell–Varley models were fitted to calculate the predatory function response, search efficiency, intraspecific interference, and predation preference of O. strigicollis on second-instar larvae and adults of M. usitatus. Our results showed that the predatory ability of O. strigicollis on second-instar larvae of M. usitatus was 78.62, and the theoretical maximum daily predation was 76.92. The predatory ability of O. strigicollis on adults of M. usitatus was 52.79, and the theoretical maximum daily predation was 52.62. The daily predation of O. strigicollis was positively correlated with prey density and negatively correlated with search efficiency. The rate of predatory action on prey decreased with an increasing O. strigicollis density, while the intensity of apportioned competition increased with an increasing O. strigicollis density. The results of the prey preference experiment showed that the preference of O. strigicollis for second-instar larvae of M. usitatus was significantly higher than for adults of M. usitatus. In conclusion, the indoor experiment data can provide an effective reference for the use of O. strigicollis to control M. usitatus infestation, providing an important biological control agent for controlling M. sativa thrips.

Grasshoppers are integral to grassland ecosystems, influencing vegetation structure and productivity while participating in energy flow and material cycling. However, increased stocking rates can alter plant community composition and structure, impacting grasshoppers’ food resources and trophic niche breadth. Little is known about how grasshopper trophic niche breadth changes under different stocking rates. To address this research gap, we conducted a field experiment in the Inner Mongolian desert steppe with four stocking rate treatments (no grazing, light, moderate, and heavy grazing). We analyzed the trophic positions of three dominant grasshopper species using stable carbon (δ13C) and nitrogen (δ15N) isotopes, and determined their dietary composition via high-throughput gut content DNA barcoding sequencing. Our results showed that moderate to heavy grazing significantly altered the isotopic values of grasshoppers, thereby affecting their trophic niche breadth and dietary preferences. Oedaleus asiaticus and Myrmeleotettix palpalis increased in abundance with higher stocking rates, despite reduced dietary diversity, likely due to their ability to exploit altered plant resources via expanded trophic niches. In contrast, Calliptamus abbreviatus experienced a decline in abundance, showing both reduced dietary diversity and no compensatory niche expansion. These findings indicate that the survival impacts of reduced dietary diversity and limited preferred resources are context-dependent, varying with species-specific adaptive capacities to grazing-induced changes. Our findings highlight that stocking rates differentially affect the dietary diversity of the three grasshopper species, with specific implications for grassland management: by optimizing stocking rates, we can regulate grasshopper population dynamics through their dietary preferences, thereby contributing to the sustainable management of desert steppe ecosystems.

BACKGROUND:Grasshoppers play a keystone role in health and stability in grassland ecosystems, which are critical for biodiversity and ecological services. Despite this, in the context of the desert steppe, there remains a paucity of knowledge regarding the response of Calliptamus abbreviatus to variations in grazing intensity and the subsequent impact on its gut microbiome and dietary diversity. The present study evaluated the impact of variations in sheep grazing intensity on the composition of plant species, which in turn influences the diet and intestinal microbiota of the grasshopper C. abbreviatus, an abundant species in these ecosystems. RESULTS:We found that increased grazing intensity has significant impacts on plant species composition and diversity, reduces grasshopper dietary diversity, increases reliance on Allium tenuissimum, and alters gut microbial community structure. Notably, plant species composition changes associated with high grazing intensity decrease Actinobacteria and Frankia but increase Proteobacteria, potentially affecting grasshopper adaptability and nutrient absorption. Procrustes analysis revealed a significant congruence between diet and gut microbiota, and the plant species changes associated with grazing serve as a pivotal point in the enterotype differentiation of grasshoppers, with enterotype 2 potentially endowing grasshoppers with enhanced nutrient absorption and transport capabilities under grazing, helping them adapt to environmental changes. CONCLUSION:Our findings emphasize that the diet and structure and function of grasshopper gut microbial communities are altered by the quality and quantity of food resources. This study provides insights into the long-term interactions between insects and their environment, and these insights are crucial for sustainable grassland management and pest control strategies. © 2025 Society of Chemical Industry.

Allium mongolicum Regel leaf beetle, Galeruca daurica (Coleoptera: Chrysomelidae) is rampantly harmful in Inner Mongolia grassland. However, the current management strategies for this pest still heavily rely on chemical control using traditional insecticides or those with novel action. In the study, we conducted an indoor bioassay to evaluate the sublethal effects of chlorantraniliprole on the biological characteristics. Additionally, we assessed the activity of detoxification enzymes, specifically the primary enzymes carboxylesterase (CarE) and cytochrome P450, and insect's secondary metabolic enzymes, namely glutathione S-transferase (GST) and uridine diphosphate glycosyltransferase (UGT), as well as its gene expression profile. The developmental period of the 3rd instar larvae of G. daurica was significantly prolonged after treatment with chlorantraniliprole, which negatively affected the hatching, pupation and diapause rates as well as their body weight. The larvae showed different dynamics of the enzyme activities within 24 h of chlorantraniliprole treatment. Different sublethal concentrations of chlorantraniliprole showed an inducing effect on the activities, while no significant difference in activity was observed for P450. Expression profiling of detoxifying enzyme genes screened by transcriptome data has revealed that 6 CarE, 2 GST, 7 UGT and 31 P450 genes were up-regulated. In conclusion, chlorantraniliprole's sublethal effect on 3rd instar G. daurica larvae resulted in a deceleration of their growth and developmental processes and significantly increased the activity levels of CarE, GST and UGT. The detoxifying proteins encoded by up-regulation genes may involve in the detoxification metabolism of chlorantraniliprole. Our results provide a basis for understanding the molecular mechanisms of chlorantraniliprole action and detoxification in this key grassland insect pest.

This study identified genetic mutations linked to resistance to pyrethroid insecticides in the plant pest Lygus pratensis. The voltage-gated sodium channel (VGSC) gene was cloned, revealing two mutations (Met918Thr and Leu1014Phe) in laboratory strains and field populations from Inner Mongolia, resulting in variable pyrethroid resistance. A 3D model of LpVGSC was created using homology modeling, and pyrethroid binding patterns were analyzed via molecular docking. Molecular dynamics simulations confirmed structural stability changes and binding stability of pyrethroids to VGSC sites. Mutation frequencies of homozygous and heterozygous genotypes did not exceed 40 and 20%, respectively. Toxicity tests showed high resistance to λ-cyhalothrin (LC50:401.31 ng/cm2). The kdr (L1014F) and superkdr (M918T) mutations weakened interaction forces, reducing pyrethroid binding. M918T and L1014F mutations are predicted to reduce Type I pyrethroid affinity, suggesting Type II pyrethroids may be more effective against resistant strains. These findings aid in resistance management and insecticide design.

NADPH-cytochrome P450 reductase (CPR) is crucial for the detoxification process catalysed by cytochrome P450, which targets various exogenous xenobiotics, as well as pesticides. In our research, we successfully obtained the complete cDNA sequence of Apolygus lucorum's CPR (AlCPR) using reverse transcription PCR along with rapid amplification of cDNA ends technology. Bioinformatics analysis exhibited that the inferred amino acid sequence of AlCPR is characteristic of standard CPRs, featuring an N-terminal membrane anchor and three conserved FMN, FAD and NADP binding sites. Phylogenetic result revealed that AlCPR was positioned within the Hemiptera cluster, showing a close evolutionary relationship with the CPR of Cimex lectularius. The real-time quantitative PCR results demonstrated widespread expression of AlCPR across various life stages and tissues of A. lucorum, with the most prominent expression in adults and the abdominal region. Injecting double-stranded RNA of AlCPR only significantly increased the lambda-cyhalothrin susceptibility in lambda-cyhalothrin-resistant strain rather than the susceptible strain. These findings suggest a potential link between AlCPR and the P450-dependent defence mechanism against lambda-cyhalothrin in A. lucorum.

The aim of the study was to clarify the trapping effects of the sticky trap on Lygus pratenss in alfalfa field. Here, eight different color sticky boards, two different hanging heights of 20 cm and 50 cm, two different hanging locations of the edge and the central of alfalfa field, and four different hanging times in the morning（7:00-11:00）,noon（11:00-15:00）,afternoon（15:00-19:00） and evening（19:00-7:00 next day） were set in the growing alfalfa field in 2021 and 2022,respectively, to investigate the attracting effects of different color sticky boards and hanging methods on collecting number of the tarnished L. pratensis in alfalfa field. The results showed that the green sticky board had the best trapping effect towards L. pratensis,the average trapping number were 9.41 and 10.79 individuals per board in 2021 and 2022 respectively, significantly higher than those from the other seven color boards. The trapping effect of green board hanged at the 50 cm height was significantly higher than that at the 20 cm（P ＜ 0.05）. The trapping effect was better when the sticky traps were set at the margin of the alfalfa field than that in the central section（P＜0.05）. The number of L. pratensis individuals trapped at afternoon was more than those at other times within the same one trapping day. In the year, it was found out that the damage of L. pratensis appeared to its peak in late May and late June; the most L. pratensis attracted in the second crop of alfalfa; and the number of L. pratensis population were the highest in the budding stage of the first and second crop.