Search Results (484)

Genetically modified crops that produce insecticidal proteins from Bacillus thuringiensis (Bt) are currently the most efficient and safest method of pest control worldwide. However, the prolonged planting period has led to a reduction in the efficacy of Bt crops due to the evolution of pest resistance in the field. This review paper examines the resistance status of lepidopteran pests to Bt crops under field conditions, elucidates the molecular mechanism underlying their resistance to Bt Cry toxins, and discusses resistance management strategies based on these mechanisms. Extensive research has demonstrated that mutations and alterations in expression patterns of midgut receptor genes are closely associated with Bt resistance. As our understanding of molecular mechanisms progresses, several innovative approaches such as DNA molecular detection techniques, engineering modified Cry toxins, and combining Bt toxin with RNAi technology have been developed for effective pest control measures. Future research will further unravel the intricate molecular mechanisms underlying this phenomenon to develop scientifically sound integrated pest management strategies. Full article

The primary mode of transmission for Chagas disease is vector-borne transmission, spread by hematophagous insects of the Triatominae subfamily. In Mexico, the triatomine Meccus pallidipennis is particularly significant in the transmission of Trypanosoma cruzi. This study focused on analyzing protein expression and modifications by glycosylation in different regions of the digestive tract of fifth-instar nymphs of M. pallidipennis. Two gut sections were dissected and extracted: the anterior midgut (AMG) and the proctodeum or rectum (RE). Proteins were extracted from each tissue sample and profiled by one- and two-dimensional electrophoresis; protein glycosylation was analyzed by lectin affinity. Our results showed significant differences in protein expression and glycosylation between both gut regions, with modifications being more frequent in the RE. The proteins HSP70, actin, and tubulin were analyzed, finding a differential expression of the latter two between AMG and RE. Understanding glycosylation patterns provides critical insights into vector–pathogen interactions that could eventually inform novel control approaches. Furthermore, the potential use of lectins as insecticidal agents highlights the broader implications of glycoprotein research in the future development of strategies on vector control to disrupt T. cruzi transmission. Full article

Public acceptance of genetically modified crops engineered with Bacillus thuringiensis (Bt) insecticidal protein genes (BT-GMCs), which confer resistance to various lepidopteran insect pests, is generally lacking. As a major concern over BT-GMCs is the allergenicity of insecticidal proteins, alleviating safety concerns should help increase public acceptance. In this study, three lepidopteran-specific Bt toxins, Cry1Aa, Cy1Ab, and Cry1Ac, were treated with simulated digestive fluids under various conditions. Western blotting using antiserum raised against individual segments (α-helices of domain I and β-sheets of domains II and III) of Cry1Aa showed that digestion produces a variety of polypeptides. In particular, the transmembrane α4–α5 of domain I, which may retain the ability to form pores, was the most resistant to digestion. Intact Cry1A toxins and these digests were then applied to RBL-2H3 cultured rat mast cells to determine whether the toxins directly induce histamine release. However, fluorescence microscopy revealed no specific binding of Cry1A toxins to RBL-2H3 cultured rat mast cells. In addition, neither the OPA method nor HPLC analysis detected significant histamine release from mast cells treated with Cry1A toxins and these digests. Our results provide important data supporting the safety of Cry1A toxins and potentially BT-GMCs. Full article

Cowpea (Vigna unguiculata) is a vital crop in sub-Saharan Africa, but the legume pod borer (LPB), Maruca vitrata, can cause over 80% yield losses. Natural resistance to this lepidopteran pest is absent in cowpea germplasm, and insecticides are ineffective due to the pest’s cryptic behavior. To address this, a genetically modified (GM) cowpea expressing the cry1Ab protein from Bacillus thuringiensis (Bt) was developed, providing complete LPB resistance. This Bt cowpea, commercialized as Sampea 20-T in Nigeria, was recently approved in Ghana as Songotra T. To evaluate its performance and the financial returns of its cultivation, field trials were conducted across multiple locations in northern Ghana to compare it to the non-transgenic Songotra control and two commercial cultivars, Kirkhouse-Benga and Wang-Kae. Songotra T exhibited protection against LPB infestations and damage, achieving a grain yield of 2534 kg/ha compared to 1414–1757 kg/ha for the other entries. As expected, non-LPB pest infestations and damage were similar across all entries. Economic analysis revealed that Songotra T had the highest return on investment (464%), outperforming the other tested cultivars (214%). These results demonstrate the potential of GM crops to enhance yields and profitability for resource-poor farmers, underscoring the value of biotechnology for addressing critical agricultural challenges. Full article

Acephate is an organophosphate foliar and soil insecticide that is used worldwide. In this study, the transgenerational ovarian developmental toxicity caused by acephate, along with its in utero reprogramming mechanisms, were explored. Thirty female virgin Wistar albino rats were randomly assigned to three groups: one control group and two acephate treatment groups. The treatment groups received daily low or high doses of acephate (34.2 mg/kg or 68.5 mg/kg body weight, respectively) from gestational day 6 until spontaneous labor, resulting in F1 offspring. At 28 days, a subgroup of F1 females were euthanized. The ovaries were extracted, thoroughly cleaned, and weighed before being fixed for further analysis. The remaining F1 females were mated with normal males to produce the F2 generation. The F1 female offspring presented reduced fertility and body weight, whereas the ovarian weight index and sex ratio increased in a dose-dependent manner. Structural analysis revealed altered follicular abnormalities with ovarian cells displaying pyknotic nuclei. Additionally, the gene and protein expression of Cyp19 decreased, whereas that of Gdf-9 increased in the high-dose treatment group (68.5 mg/kg). We also observed significantly increased expression levels of ovarian estrogen receptor 1 (Esr1) and insulin-like growth factor 1 (Igf1), whereas Insl3 expression was significantly decreased. The F2 female offspring presented reproductive phenotype alterations similar to those of F1 females including decreased fertility, reduced Cyp19 gene and protein expression, and structural ovarian abnormalities similar to those of polycystic ovary syndrome (PCOS). In conclusion, acephate induced ovarian developmental toxicity across two generations of rats, which may be linked to changes in the ovarian Cyp19, Gdf9, Insl3, and Igf1 levels. Full article

Developing simple and efficient multi-gene expression systems is crucial for multi-trait improvement or bioproduction in transgenic plants. In previous research, an IGG6-based bicistronic system from the nonpathogenic fungus Glarea lozoyensis efficiently expressed multiple enzyme proteins in yeast and maize, and the heterologous enzymes successfully performed their catalytic activity to reconstruct the biosynthetic pathway in the host organism. Unlike enzyme proteins, some heterologous functional proteins (such as insecticidal proteins) are dose-dependent and they need to express sufficient levels to perform their biological functions. It remains unclear whether the IGG6-based bicistronic system can achieve high expression of the functional proteins for practical applications in crops. In this study, two Bacillus thuringiensis (Bt) insecticidal genes, vip3Aa and cry1Ab, were linked via IGG6 to form a bicistron, while two glyphosate resistance genes, gr79epsps and gat, served as monocistronic selectable marker genes. Regenerated maize plants were produced through genetic transformation. RNA and immunoblot analyses revealed that the vip3Aa-IGG6-cry1Ab bicistron was transcribed as a single transcript, which was then translated into two separate proteins. Notably, the transcription and translation of cry1Ab were significantly positively correlated with those of vip3Aa. Through ELISA and leaf bioassay, we identified two transgenic maize lines, VICGG-15 and VICGG-20, that exhibited high insecticidal activity against fall armyworm (FAW; Spodoptera frugiperda) and Asian corn borer (ACB; Ostrinia furnacalis), both of which had high expression of Vip3Aa and Cry1Ab proteins. Subsequent evaluations, including silk, ear, and field bioassays, as well as glyphosate tolerance assessments, indicated that the VICGG-15 plants displayed high resistance to FAW and ACB, and could tolerate up to 3600 g acid equivalent (a.e.) glyphosate per hectare without adversely affecting phenotype or yield. Our finding established that the IGG6-based bicistronic system can achieve high expression of functional proteins in maize, and it is a potential candidate for multi-gene assembly and expression in plants. Full article

Insecticides are used commonly in agricultural production to defend plants, including legumes, from insect pests. It is a known fact that insecticides can have a harmful effect on the legume–rhizobial symbiosis. In this study, the effects of systemic seed treatment insecticide Imidor Pro (imidacloprid) and foliar insecticide Faskord (alpha-cypermethrin) on the structural organization of pea (Pisum sativum L.) nodules and their transcriptomic activity were investigated. The plants were treated as recommended by the manufacturer (10 mg/mL for Imidor Pro and 50 µg/mL for Faskord) and twofold concentrations were used for both insecticides. Insecticides had no visible effect on the growth of pea plants. The nodules also showed no visible changes, except for the variant treated with twofold concentration of Imidor Pro. However, the dry weight of shoots and roots differed significantly in insecticide-treated plants compared to untreated plants in almost all treatments. The number of nodules decreased in variants with Imidor Pro treatment. At the ultrastructural level, both insecticides caused cell wall deformation, poly-β-hydroxybutyrate accumulation in bacteroids, expansion of the peribacteroid space in symbiosomes, and inclusions in vacuoles. Treatment with Faskord caused chromatin condensation in nucleus. Imidor Pro treatment caused hypertrophy of infection droplets by increasing the amount of matrix, as confirmed by immunofluorescence analysis of extensins. Transcriptome analysis revealed upregulation of expression of a number of extensin-like protein-coding genes in nodules after the Imidor Pro treatment. Overall, both insecticides caused some minor changes in the legume–rhizobial system when used at recommended doses, but Faskord, an enteric contact insecticide, has fewer negative effects on symbiotic nodules and legume plants; of these two insecticides, it is preferred in pea agricultural production. Full article

Chrysodeixis includens and Rachiplusia nu are two species belonging to the Plusiinae subfamily within the Noctuidae family. Due to their morphological similarity, the identification of their larvae is difficult and time-consuming. A rapid and accurate identification of these two species is essential for their management as these species exhibit differential susceptibilities to insecticides and crops engineered to express Bacillus thuringiensis (Bt) proteins, and a molecular tool can easily provide this differentiation. Currently, molecular analysis can identify these species through genetic sequencing, an expensive and time-consuming process. In our study, after sequencing part of the mtDNA cytochrome c oxidase I (COI) gene and based on the differences found in the gene of each species, a set of species-specific primers was developed: one reverse primer common to both species and two forward primers, specific to each species, amplifying fragments of 199 base pairs (bp) for C. includens and 299 bp for R. nu. Our results indicate that the primers were specific for these species, enabling the identification of individuals directly through agarose gel. The new methodology proved to be accurate, rapid, and reliable for the correct identification of these two species of loopers. Full article

The growing emphasis on food safety and healthier lifestyles, driven by industrial expansion and scientific priorities, has highlighted the necessity of managing harmful microorganisms to guarantee food quality. A significant challenge in this domain is the control of pathogens that are capable of forming biofilms, entering a sessile state that enhances their resistance to broad-spectrum antibiotics. Essential oils, renowned for their antibacterial properties, present a promising natural alternative for food preservation. In this study, we analyzed the chemical composition of Santalum album essential oil (SAEO) using GC-MS, identifying (Z)-α-santalol (57.1%) as the primary constituent. Antimicrobial activity was confirmed through disc diffusion and minimum inhibitory concentration (MIC) assays against Gram-positive and Gram-negative bacteria and yeast from the genus Candida. Additionally, in situ experiments demonstrated that vapor-phase SAEO effectively inhibited Serratia marcescens on the food model, supporting its potential as a natural preservative. MBIC assays, crystal violet staining, and MALDI-TOF MS analysis on S. enterica biofilms were used to further evaluate the antibiofilm effects of SAEO. The crystal violet assay revealed a strong antibiofilm effect, while the MALDI-TOF MS analysis showed changes in the bacterial protein profiles on both glass and plastic surfaces. SAEO also showed significant anti-Salmonella activity on vacuum-packed carrot slices. SAEO outperformed the control samples. The insecticidal activity against Megabruchidius dorsalis was also studied in this work, and the best insecticidal activity was found at the highest concentrations. These findings indicate that SAEO could serve as a valuable component in food preservation, with notable antibacterial and antibiofilm benefits. Full article

Oulema melanopus L., 1758 (Coleoptera: Chrysomelidae) is one of the significant pests affecting cereal crops in Europe. Its damage is evident in the destruction of leaves during the spring growing season, leading to substantial impacts on both the quantity and quality of the harvested yields. The study aimed to evaluate the extent of leaf surface damage, changes in chlorophyll content caused by this pest, and the subsequent effects on yield quality. To achieve this, two experimental parcels were established, each subjected to different pesticide treatments during the spring vegetation cycle, but notably, with the difference that one parcel did not receive insecticide applications. The phytosanitary status, yield quantity, and quality parameters of thes parcels were compared. Chlorophyll content in damaged and undamaged plants was monitored in vivo using SPAD measurements, while the extent of leaf surface damage was assessed through image analysis using GIMP software 2.10.32. Harvested grain underwent milling and baking analysis, with milling and baking-quality indicators measured using a NIR grain analyzer. The results revealed that omitting springtime insecticide treatments during the emergence of O. melanopus led to significant reductions in leaf area and yield quality. In untreated parcels, leaf decession followed linear progression, reaching 35–40% within 20 days. This damage correlated with the decline in SPAD index values, indicating a 40–50% reduction in chlorophyll content dependent photosynthetic activity. Consequently, there were substantial decreases in milling and baking qualities, including nearly 30% reductional protein-content indicators and 10% in the Hagberg falling number. In summary, our large-scale field experiments demonstrated that persistent O. melanopus damage in wheat fields significantly reduced both the quantity and quality of yields, particularly protein content. These facts underscore the economic importance of timely pest-control measures to mitigate damage and preserve crop value. Full article

The objective of this study was to identify the Hsp70s in Paracoccus marginatus and explore their roles in P. marginatus’s resistance to temperature and insecticide stress. The full-length cDNA sequences of PmHsp70s were obtained by PCR cloning and sequencing. The physicochemical and structural characteristics of PmHsp70s were analyzed, and a phylogenetic tree was constructed. The gene expressions of PmHsp70s were detected using qRT-PCR to explore the impacts of temperature and insecticide stress on P. marginatus. A total of 12 PmHsp70s were identified and cloned. The amino acids encoded by PmHsp70s were found to contain highly conserved regions characteristic of the Hsp70 family. The subcellular localization results showed that the majority of PmHsp70s were located in the cytoplasm. A total of 13 unique conserved motifs were identified for the PmHsp70s, of which 9 were shared motifs. The phylogenetic tree showed that the 12 PmHsp70s could be clustered into five branches, with the closest evolutionary relationship observed with the Phenacoccus solenopsis. The expression of the majority of PmHsp70s was up-regulated in P. marginatus when subjected to heat stress, with the higher expression fold change observed for PmHsp70-9, PmHsp70-11, and PmHsp70-12. The expression of specific PmHsp70s was notably suppressed under cold stress, whereas the expression of others was markedly enhanced. Upon exposure to chlorfenapyr and lambda-cyhalothrin, the expressions of PmHsp70-11 and PmHsp70-12 were significantly up-regulated with the highest expression fold change, respectively. The results revealed the significance of specific PmHsp70s in the resistance of P. marginatus to temperature and insecticide stress. This study improved our understanding of the mechanisms underlying P. marginatus’s adaptive responses to unfavorable environmental conditions. Full article

Enhanced expression of carboxylesterase (CarE) genes is an important mechanism of insecticide resistance in pests. However, their roles in multi-insecticide resistance have rarely been reported. Herein, two CarE genes (PxαE6 and PxαE9) were identified; their relative expression levels in three multi-insecticide-resistant populations of P. xylostella (HN, GD-2017 and GD-2019) were 2.69- to 15.32-fold higher than those in the sensitive population, and they were considerably overexpressed at the larval stage and in the midgut of the 4th instar. PxαE6 and PxαE9 knockdown increased the susceptibility of GD-2019 larvae to phoxim or/and beta-cypermethrin. The recombinant PxαE6 and PxαE9 expressed in Escherichia coli exhibited high hydrolysis activity towards α-NA. GC–MS and LC–MS/MS assays revealed that PxαE9 could metabolize beta-cypermethrin and phoxim with efficiency determinations of 51.6% and 21.1%, respectively, while PxαE6 could metabolize phoxim with an efficiency of 12.0%. Homology modelling, molecular docking and molecular-dynamics simulation analyses demonstrated that beta-cypermethrin or/and phoxim could fit well into the active pocket and stably bind to PxαE6 or PxαE9. These results show that PxαE6 and PxαE9 overexpression were involved in resistance to beta-cypermethrin or/and phoxim in multi-insecticide-resistant P. xylostella populations, a finding which sheds light on the molecular mechanisms of multi-insecticide resistance in insect pests. Full article

Cry2Ab is a significant alternative Bacillus thuringiensis (Bt) protein utilized for managing insect resistance to Cry1 toxins and broadening the insecticidal spectrum of crops containing two or more Bt genes. Unfortunately, the identified receptors fail to fully elucidate the mechanism of action underlying Cry2Ab. Previous studies have demonstrated the involvement of vacuolar H⁺-ATPase subunits A, B, and E (V-ATPase A, B, and E) in Bt insecticidal activities. The present study aims to investigate the contribution of V-ATPase C to the toxicities of Cry2Ab against Helicoverpa armigera. The feeding of Cry2Ab in H. armigera larvae resulted in a significant decrease in the expression of V-ATPase C. Further investigations confirmed the interaction between V-ATPase C and activated Cry2Ab protein according to Ligand blot and homologous and heterologous competition assays. Expressing endogenous HaV-ATPase C in Sf9 cells resulted in an increase in Cry2Ab cytotoxicity, while the knockdown of V-ATPase C by double-stranded RNAs (dsRNA) in midgut cells decreased Cry2Ab cytotoxicity. Importantly, a higher toxicity of the mixture containing Cry2Ab and V-ATPase C against insects was also observed. These findings demonstrate that V-ATPase C acts as a binding receptor for Cry2Ab and is involved in its toxicity to H. armigera. Furthermore, the synergy between V-ATPase C protein and Cry2Ab protoxins provides a potential strategy for enhancing Cry2Ab toxicity or managing insect resistance. Full article

Cry toxins from Bacillus thuringiensis are effective biopesticides that kill lepidopteran pests, replacing chemical pesticides that indiscriminately attack both target and non-target organisms. However, resistance in susceptible pests is an emerging problem. B. thuringiensis also produces vegetative insecticidal protein (Vip3A), which can kill insect targets in the same group as Cry toxins but using different host receptors, making the combined application of Cry and Vip3A an exciting possibility. Vip3A toxicity requires the formation of a homotetramer. Hence, screening of Vip3A mutants for increased stability requires orthogonal biophysical assays that can test both tetrameric integrity and monomeric robustness. For this purpose, we have used herein for the first time a combination of analytical ultracentrifugation (AUC), mass photometry (MP), differential static light scattering (DSLS) and differential scanning fluorimetry (DSF) to test five mutants at domains I and II. Although all mutants appeared more stable than the wild type (WT) in DSLS, mutants that showed more dissociation into dimers in MP and AUC experiments also showed earlier thermal unfolding by DSF at domains IV–V. All of the mutants were less toxic than the WT, but toxicity was highest for domain II mutations N242C and F229Y. Activation of the protoxin was complete and resulted in a form with a lower sedimentation coefficient. Future high-resolution structural data may lead to a deeper understanding of the increased stability that will help with rational design while retaining native toxicity. Full article

The use of egg parasitoids in Augmentative Biological Control (ABC) is a highly effective strategy within the integrated pest management (IPM) of lepidopteran defoliators. Safer than chemical insecticides, these natural antagonists have demonstrated significant efficacy. Trichogramma pretiosum and Telenomus remus, known for their high parasitism rates, are the most extensively used and studied parasitoids for controlling economically important lepidopterous in crops such as soybean and maize. Brazil, a leading adopter of crops expressing Bacillus thuringiensis (Bt) proteins, faces growing field-evolved resistance to Cry proteins in soybean and maize. This resistance, particularly of Rachiplusia nu in soybean and Spodoptera frugiperda in maize, has become more prominent in recent years, increasing insecticide use. Therefore, this article reviews the current status of egg parasitoids adoption in ABC against lepidopteran pests, emphasizing the role of Tr. pretiosum and the potential of Te. remus as sustainable alternatives to chemical insecticides to manage pests in both non-Bt and Bt crops. Additionally, we provide recommendations for using these parasitoids in ABC programs and discuss the challenges that must be addressed to optimize the adoption of biocontrol agents in ABC programs for maximum benefit. Full article