Search Results (100)

The insect resistance expression of Bacillus thuringiensis (Bt) cotton (Gossypium hirsutum L.) is unstable due to temporal and spatial variations in the Bt protein content in different organs and growth stages. The aim of this study was to improve the Bt protein content in cotton flowers and investigate the underlying physiological mechanism using biochemical analytical methods. In this study, a split-plot design with three replications was used. The main plots included two Bt cotton cultivars (a conventional cultivar, Sikang1 (S1), and a hybrid cultivar, Sikang3 (S3)), while five soil nitrogen application levels (CK (control check): normal level; N1: 125% of the CK; N2: 150% of the CK; N3: 175% of the CK; N4: 200% of the CK) constituted the subplots. The Bt protein content and related nitrogen metabolism parameters were measured. We found that the Bt protein content increased and then decreased with increasing nitrogen rates. It reached its maximum at N3, with significant increases of 71.86% in 2021 and 39.36% in 2022 compared to the CK. Correlation analysis indicated that the Bt protein content was significantly positively related to the soluble protein and free amino acid contents, as well as the GPT (glutamic pyruvic transaminase), GOT (glutamic oxaloacetic transaminase), GS (glutamine synthetase) and GOGAT (glutamate synthetase) activities. On the other hand, negative correlations were found between the Bt protein content and protease and peptidase activities. In addition, stepwise regression and path analysis indicated that the increased Bt protein content was mainly due to the enhanced GS and GOGAT activities. In summary, appropriately increasing nitrogen fertilizer application is a practical way to increase flower Bt protein content and insecticidal efficacy of Bt cotton. These findings provide an actionable agronomic strategy for sustaining Bt expression during the critical flowering period. Full article

Globally, insect pests adversely affect approximately 75% of the most important crops. However, the widespread use of chemical insecticides has significant drawbacks, including non-specific biological activity, toxicity to humans, detrimental effects on beneficial insects, and the rapid development of resistance. In this context, prodigiosin—a tripyrrolic secondary metabolite produced by various microorganisms—emerges as a promising alternative due to its favourable properties, such as being non-toxic, environmentally safe, non-irritant, and non-allergenic, and having non-carcinogenic potential. Prodigiosin has demonstrated insecticidal efficiency against pests at various developmental stages. Studies suggest that prodigiosin inhibits enzymes like acetylcholine esterase, protease, and acid phosphatase and induces oxidative stress. This review explores the potential of prodigiosin as an eco-friendly insecticide, discussing its production, extraction, and purification processes and its advantages, disadvantages, and mechanism of action, and future perspectives. Special emphasis is given to using non-pathogenic strains to mitigate biosafety concerns. Full article

Serine proteases are widely distributed in both invertebrates and vertebrates, playing critical roles in the regulation of innate immunity. In the insect innate immune system, two pivotal pathways—the prophenoloxidase (PPO) activation cascade and Toll pathway-mediated antimicrobial peptide (AMP) synthesis—are both tightly regulated by serine protease cascades. This study focuses on serine protease–hemolymph protease 6 of A. pernyi (Ap-HP6). Following immune stimulation, the expression of Ap-proHP6 was significantly induced, primarily observed in hemocytes and the fat body. After suppressing Ap-proHP6 expression via RNA interference (RNAi) and infecting larvae with different microbes, the expression levels of AMPs showed a downward trend. When endogenous Ap-proHP6 content in hemolymph was reduced using RNAi technology or anti-rAp-proHP6-His₆ polyclonal antibodies, PAMPs/microbe-mediated phenoloxidase (PO) activity significantly decreased. These results suggest that Ap-HP6 has a positive regulatory effect on PPO activation and AMP synthesis. Additionally, the in vitro hydrolysis of rAp-proHP6-Tb-His₆ yielded rAp-HP6 with serine protease activity, which exhibited optimal reaction conditions for S-2288 at pH 8.0, 50 °C, and 15 min. Full article

The red palm weevil, Rhynchophorus ferrugineus, is a destructive, invasive pest to a diverse range of palm plantations globally. Commonly used broad-range chemical insecticides for insect control pose high risks to non-target organisms, humans, and the environment. A bio-rational approach of screening natural small-molecule inhibitors that specifically target R. ferrugineus proteins critical to its life processes can pave the way for developing novel bioinsecticides. Digestive enzymes (DEs), which impair feeding on plants (herbivory), are promising targets. We generated de novo transcriptomes, annotated DE-related genes from the R. ferrugineus gut and abdomen, manually annotated the DE gene family from the recently available genome and our transcriptome data, and reported 34 glycosidases, 85 lipases, and 201 proteases. We identified several tandem duplicates and allelic variants from the lipase and protease families, notably, 10 RferLip and 21 RferPro alleles, which emerged primarily through indels and single-site substitution. These alleles may confer enhanced digestive lipolysis and proteolysis. Phylogenetic analyses identified and classified different subfamilies of DEs and revealed close evolutionary relationships with other coleopterans. We assessed select candidate DEs’ activity and the potential for inhibition in silico to better understand the herbivory arsenal. In silico analysis revealed that the selected enzymes exhibited similar ligand-binding affinity to their corresponding substrate, except for protease aminopeptidase N, RferPro40, which exhibited poorer affinity to the inhibitor bestatin. Overall, our study serves as a foundation for further functional analysis and offers a novel target for the development of a novel bio-rational insecticide for R. ferrugineus. Full article

This review provides an in-depth examination of the intricate interactions between plant metabolites and the digestive and antioxidative enzymes in insects, highlighting their essential roles in shaping insect herbivory and adaptation strategies. Plants have evolved a diverse arsenal of secondary metabolites to defend against herbivorous insects, which, in response, have developed sophisticated adaptations to overcome these defenses and efficiently exploit plant resources. We outline the importance of digestive enzymes, such as proteases and amylases, which allow insects to break down complex plant compounds and access vital nutrients. Additionally, the review focuses on antioxidative enzymes in the insect midgut, including superoxide dismutase and catalase, which play a crucial role in mitigating oxidative stress generated during digestion and other metabolic processes. Synthesizing findings from various studies, this review also considers how environmental factors, such as heavy metal exposure and temperature changes, influence these enzymes’ activity levels. It highlights the dual function of antioxidative enzymes in detoxifying harmful plant-derived compounds while preserving cellular stability. The implications of these biochemical interactions for pest management are discussed, with an emphasis on the potential for developing biopesticides that target specific enzymatic pathways to disrupt insect feeding and growth. By elucidating the biochemical mechanisms that underlie plant-insect interactions, this review enhances our understanding of co-evolutionary dynamics and offers insights into sustainable agricultural practices that could leverage these interactions for effective pest control. Finally, the review proposes future research directions aimed at identifying novel plant metabolites with enzyme-modulating properties and exploring the ecological impacts of enzyme-targeted pest management approaches. Full article

The red palm weevil (RPW; Coleoptera: Curculionidae) is a destructive pest affecting palms worldwide, capable of causing significant economic losses and ecological damage in managed palm ecosystems. Current management heavily relies on synthetic insecticides, but their overuse fosters resistance. Bacillus thuringiensis (Bt) offers a promising alternative, producing toxins selective against various insect orders, including Coleoptera. However, no specific Bt toxin has yet been identified for RPW. This study investigates the toxicity against RPW larvae of the Bt Cry1Ia protoxin, known for its dual activity against Lepidoptera and Coleoptera. A laboratory RPW colony was reared for two generations, ensuring a reliable insect source for bioassays. Cry1Ia was expressed as a 6xHis-tagged fusion protein in Escherichia coli and purified using nickel affinity. Incubation with RPW larval gut proteases for 24 h produced a stable core of ~65 kDa. Diet-incorporation bioassays revealed high Cry1Ia toxicity in neonate larvae. In contrast, the lepidopteran-active Cry1Ac protoxin, used as a robust negative control, was completely degraded after 24 h of in vitro proteolysis and showed no toxicity in bioassays. Cry1Ia-fed larvae exhibited significant midgut cell damage, characteristic of Bt intoxication. These findings highlight Cry1Ia’s strong potential for integration into RPW management programs. Full article

Protease inhibitors are biomolecules with growing biotechnological and biomedical relevance, including those derived from plants. This study investigated strong trypsin inhibitors in quinoa, amaranth, and lupine seeds, plant grains traditionally used in Andean South America. Amaranth seeds displayed the highest trypsin inhibitory activity, despite having the lowest content of aqueous soluble and thermostable protein material. This activity, directly identified by enzymatic assay, HPLC, intensity-fading mass spectrometry (IF-MS), and MS/MS, was attributed to a single protein of 7889.1 Da, identified as identical in Amaranthus caudatus and A. hybridus, with a K_i of 1.2 nM for the canonical bovine trypsin. This form of the inhibitor, which is highly homogeneous and scalable, was selected, purified, and structurally–functionally characterized due to the high nutritional quality of amaranth seeds as well as its promising agriculture–biotech–biomed applicability. The protein was crystallized in complex with bovine trypsin, and its 3D crystal structure resolved at 2.85 Å, revealing a substrate-like transition state interaction. This verified its classification within the potato I inhibitor family. It also evidenced that the single disulfide bond of the inhibitor constrains its binding loop, which is a key feature. Cell culture assays showed that the inhibitor did not affect the growth of distinct plant microbial pathogen models, including diverse bacteria, fungi, and parasite models, such as Mycoplasma genitalium and Plasmodium falciparum. These findings disfavour the notion that the inhibitor plays an antimicrobial role, favouring its potential as an agricultural insect deterrent and prompting a redirection of its functional research. Full article

This study explores the potential of Tenebrio molitor protein hydrolysates as functional food ingredients, evaluating their bioactivity and consumer acceptance of the incorporation of edible insects into food across Poland and Spain. By aligning technical advancements with consumer preferences, this research bridges the gap between laboratory innovation and market feasibility, contributing to the development of sustainable functional foods. The study optimized the process of enzyme hydrolysis using serine protease from Cucurbita ficifolia, thereby enhancing DPPH scavenging capacity increased from 3.15 ± 0.53 to 8.17 ± 0.62 µM Trolox/mL and ABTS decolorization capacity increased from 4.29 ± 0.01 to 10.29 ± 0.01 µM Trolox/mL after 5 h of hydrolysis. Consumer surveys incorporating the Food Neophobia, Insect Phobia, and Entomophagy Scales revealed demographic and cultural influences on entomophagy acceptance. Among respondents, 27.1% in Poland and 25.7% in Spain had previously consumed insect-based products, while Polish participants showed a higher willingness to adopt insect-enriched foods. The study confirmed that hydrolysis enhances the antioxidant activity of T. molitor protein hydrolysates and that demographic and cultural factors significantly influence consumer acceptance of insect-based foods. Full article

Adipocytes secrete adipokines, bioactive molecules crucial for various physiological processes, such as enhancing insulin sensitivity, promoting wound healing, supporting hair growth, and exhibiting anti-aging effects on the skin. With the growing global demand for sustainable and alternative protein sources, insect-derived proteins, particularly from Tenebrio molitor (mealworms), have gained attention due to their high nutritional value and functional bioactivities. This study aims to explore the potential of mealworm-derived protein hydrolysates as novel bioactive materials for promoting adipogenesis and improving adipokine expression, with applications in metabolic health and skin regeneration. Protein hydrolysates (<1 kDa) were prepared using enzymatic hydrolysis with three proteases (alcalase, flavourzyme, and neutrase) and evaluated for their adipogenic activity in 3T3-L1 preadipocytes. Among them, the flavourzyme-derived hydrolysate (Fh-T) exhibited the most significant effects, enhancing adipogenic differentiation and lipid accumulation. Fh-T facilitated adipogenesis by promoting mitotic clonal expansion (MCE) during the early stage of differentiation, which was associated with the upregulation of C/EBPδ and the downregulation of p27. These findings underscore the potential of mealworm-derived protein hydrolysates, particularly Fh-T, as sustainable and functional ingredients for use in glycemic control, skin health, and tissue regeneration. This study provides valuable insights into the innovative use of alternative protein sources in functional foods and cosmeceuticals. Full article

Insect foliar herbivory is ubiquitous in terrestrial ecosystems, yet its impacts on soil nitrogen cycling processes remain not yet well known. To examine the impacts of insect foliar herbivory on soil N₂O emission flux and available nitrogen (N), we conducted a pot experiment to measure soil available N content and soil N₂O emission flux among three treatments (i.e., leaf herbivory, artificial defoliation, and control,) in two broad-leaved trees (Cinnamomum camphora and Liquidambar formosana) and two conifer trees (Pinus massonianna and Cryptomeria fortunei). Our results showed that insect foliar herbivory significantly increased soil inorganic N (i.e., NH₄⁺–N and NO₃⁻–N), dissolved organic nitrogen (DON) and microbial biomass nitrogen (MBN) contents, and urease activity compared to control treatment. However, there were no differences in soil available N contents and urease activity between artificial defoliation and control treatments, implying that insect foliar herbivory had greater impacts on soil available N contents compared to physical damage of leaves. Moreover, soil N₂O emission fluxes were increased by insect foliar herbivory in Cinnamomum camphora and Pinus massonianna, but not for the other two tree species, indicating various effect of insect foliar herbivory on soil N₂O emission among tree species. Furthermore, our results showed the positive correlations between soil N₂O emission flux and soil NO₃⁻–N, DON, MBN, and acid protease activity, and soil inorganic N, pH, and MBN mainly explained soil N₂O emission. Our results implied that insect foliar herbivory can speed up soil nitrogen availability in subtropical forests, but the impacts on soil N₂O emission are related to tree species. Full article

In this study, we selected Sitona callosus, one of the primary insect pests of alfalfa, as the experimental insect and infected it with Beauveria bassiana. Transcriptomic and metabolomic analyses were conducted to explore alterations in gene expression and metabolic processes in S. callosus at 48, 96, and 144 h post infection with B. bassiana. The transcriptomic analysis unveiled that B. bassiana infection boosted immune responses in tubercula, affecting carbohydrate metabolism, cytochrome P450 activity, lysosome function, apoptosis regulation, phagosome formation, glutathione metabolism, amino acid metabolism, and pathogen response pathways. Subsequent metabolomics analysis confirmed that glycerophospholipids, carboxylic acids and derivatives, organooxygen compounds, keto acids and derivatives, and azane immune metabolites were significantly upregulated in response to B. bassiana infection. Additionally, we utilized the Pearson correlation coefficient method to examine the relationships between differentially expressed immune-related genes and metabolites, revealing notably strong correlations between these two sets of variables. By leveraging the WGCNA method to analyze immune metabolite data for immune-related genes, we identified hub genes crucial at various stages of immune activation. These central genes predominantly included C-type lectin receptors for pattern recognition, cytochrome P450 enzymes linked to detoxification processes, and cathepsin proteases. By combining transcriptome and metabolome analyses, it was determined that autophagy and arachidonic acid metabolism play significant roles in the response of S. callosus to infection by B. bassiana. This research will facilitate the understanding of the immune response to B. bassiana infection in adult S. callosus, laying a theoretical groundwork for future biological control strategies targeting S. callosus. Full article

Aphids are small, notorious insect pests that negatively impact plant health and agricultural productivity through direct damage, such as sap-sucking, and indirectly as vectors of plant viruses. Plants respond to aphid feeding with a variety of molecular mechanisms to mitigate damage. These responses are diverse and highly dynamic, functioning either independently or in combination. Understanding plant–aphid interactions is crucial for revealing the full range of plant defenses against aphids. When aphids infest, plants detect the damage via specific receptor proteins, initiating a signaling cascade that activates defense mechanisms. These defenses include a complex interaction of phytohormones that trigger defense pathways, secondary metabolites that deter aphid feeding and reproduction, lectins and protease inhibitors that disrupt aphid physiology, and elicitors that activate further defense responses. Meanwhile, aphids counteract plant defenses with salivary effectors and proteins that suppress plant defenses, aiding in their successful colonization. This review offers a detailed overview of the molecular mechanisms involved in plant–aphid interactions, emphasizing both established and emerging plant defense strategies. Its uniqueness lies in synthesizing the recent progress made in plant defense responses to aphids, along with aphids’ countermeasures to evade such defenses. By consolidating current knowledge, this review provides key insights for developing sustainable strategies to achieve crop protection and minimize dependence on chemical pesticides. Full article

The spread of invasive pests exacerbates the direct damage to host plants and the potential threat to the environment. Silicon has the potential to enhance host plant resistance to insects while also increasing plant yield. This study evaluated changes in Italian ryegrass biological yield and resistance to fall armyworm (Spodoptera frugiperda) larvae after silicon supplementation (sodium silicate and potassium silicate at 6 mmol·L⁻¹ were denoted as groups T1 and T2, respectively). Silicon supplementation significantly increased the shoot biological yield (T1 by 30.26%, T2 by 23.05%) and silicon content (T1 by 22.61% and T2 by 12.43%) of Italian ryegrass. At the same time, silicon supplementation increased the protein, soluble sugar, and vitamin contents of Italian ryegrass, while also stimulating the improvement of its physical and chemical defenses. Therefore, even though the nutrient intake of fall armyworm increased, the synergistic physical-chemical defense formed by silica deposition, flavonoid content, and increased protease inhibitor activity in the Italian ryegrass still weakened the antioxidant capacity of the larvae and inhibited larval feeding and protein accumulation. The larval body weight of the T1 and T2 groups decreased by 20.32% and 15.16%, respectively. The comprehensive scores showed that sodium silicate and potassium silicate of the same concentration had similar effects on the growth and insect resistance of Italian ryegrass. These findings suggest that both sodium and potassium silicate are effective silicon supplements for host plants. Therefore, reasonable supplementation of silicon fertilizer may become an important alternative plan for optimizing the comprehensive pest control strategy in agricultural production areas in the future, but this still needs further field research verification. Full article

Rice planthoppers, including Nilaparvata lugens, Sogatella furcifera, and Laodelphax striatellus, are major agricultural pests. Serpins, which function as serine protease inhibitors, play a pivotal role in the immune systems of these insects, especially within the Toll signaling pathway and the prophenoloxidase (PPO) cascade. This study presents a comparative analysis of serpin genes among these species, highlighting their roles in immunity and development. Utilizing genomic and bioinformatics approaches, we identified 11, 11, and 14 serpin genes in N. lugens, S. furcifera, and L. striatellus, respectively. Phylogenetic analysis revealed a close evolutionary relationship between these serpin genes and Bombyx mori BmSerpins, emphasizing the functional diversity of the serpin family. Structural analysis confirmed the presence of the reactive center loop (RCL) in all serpin proteins, with the Serpin7 subfamily showing a unique dual RCL configuration. Expression profiling showed species-specific serpin expression patterns across different life stages and adult tissues. Moreover, transcriptional analysis of serpin genes in the three planthoppers following Metarhizium infection uncovered distinct immune regulatory patterns two days post-infection. Notably, the expression of NlSerpin2-2/6, SfSerpin4/6/7-1, and LsSerpin4/5-2/6 was upregulated post-infection, potentially enhancing antifungal capabilities. In contrast, the expressions of NlSerpin1/7-1/9 and LsSerpin1/2/3/8/13 were downregulated, possibly suppressing immune responses. Moreover, Serpin6s, which share a conserved phylogenetic lineage, exhibited enhanced immune activity in response to fungal invasion. These insights into serpin-mediated immune regulation could contribute to the development of novel pest-control strategies. Full article

Jasmonic acid (JA) is a regulator of plant resistance to phytophagous insects, and exogenous JA treatment induces plant insect resistance. This study investigated the mechanism of exogenous JA-induced resistance of Hemerocallis citrina Baroni (daylily) to Thrips palmi at the biochemical and molecular levels. Daylily leaves sprayed with JA showed significantly higher levels of secondary metabolites—tannins, flavonoids, and total phenols, and activity of defense enzymes—peroxidase, phenylalanine ammonia lyase, polyphenol oxidase, and protease inhibitor (PI) than control leaves; the most significant effects were observed with 1 mmol L⁻¹ JA. Owing to an improved defense system, significantly fewer T. palmi were present on the JA-treated plants than control plants. The JA-treated leaves had a smoother wax layer and fewer stomata, which was unfavorable for insect egg attachment. The differentially expressed genes (DEGs) were significantly enriched in insect resistance pathways such as lignin and wax biosynthesis, cell wall thickening, antioxidant enzyme synthesis, PI synthesis, secondary metabolite synthesis, and defense hormone signaling. A total of 466 DEGs were predicted to be transcription factors, mainly bHLH and WRKY family members. Weighted gene co-expression network analysis identified 13 key genes; TRINITY_DN16412_c0_g1 and TRINITY_DN6953_c0_g1 are associated with stomatal regulation and lipid barrier polymer synthesis, TRINITY_DN7582_c0_g1 and TRINITY_DN11770_c0_g1 regulate alkaloid synthesis, and TRINITY_DN7597_c1_g3 and TRINITY_DN1899_c0_g1 regulate salicylic acid and ethylene biosynthesis. These results indicate that JA treatment of daylily improved its resistance to T. palmi. These findings provide a scientific basis for the utilization of JA as an antagonist to control T. palmi in daylily. Full article