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International Journal of Molecular... Aug 2019Nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome P450 reductases (CPRs) function as redox partners of cytochrome P450 monooxygenases (P450s). CPRs and...
Nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome P450 reductases (CPRs) function as redox partners of cytochrome P450 monooxygenases (P450s). CPRs and P450s in insects have been found to participate in insecticide resistance. However, the CPR of the moth has not been well characterized yet. Based on previously obtained transcriptome information, a full-length CPR cDNA of () was PCR-cloned. The deduced amino acid sequence contains domains and residues predicted to be essential for CPR function. Phylogenetic analysis with insect CPR amino acid sequences showed that is closely related to CPRs of Lepidoptera. Quantitative reverse transcriptase PCR (RT-qPCR) was used to determine expression levels of in different developmental stages and tissues of . expression was strongest at the sixth-instar larvae stage and fifth-instar larvae showed highest expression in the midgut. Expression of in the midgut and fat body was strongly upregulated when fifth-instar larvae were exposed to phoxim at LC (4 μg/mL) and LC (20 μg/mL) doses. RNA interference (RNAi) mediated silencing of increased larval mortality by 34.6% (LC dose) and 53.5% (LC dose). Our results provide key information on the gene and indicate that expression levels in larvae influence their susceptibility to phoxim and possibly other insecticides.
Topics: Amino Acid Sequence; Animals; Gene Expression Regulation; Gene Silencing; Insecticide Resistance; Insecticides; Larva; NADPH-Ferrihemoprotein Reductase; Organothiophosphorus Compounds; Phylogeny; RNA Interference; Spodoptera
PubMed: 31390813
DOI: 10.3390/ijms20153839 -
Insect Molecular Biology Jun 2020Glutathione S-transferases (GSTs) are members of a multifunctional enzyme superfamily. Forty-one GSTs have been identified in Tribolium castaneum; however, none of the...
Glutathione S-transferases (GSTs) are members of a multifunctional enzyme superfamily. Forty-one GSTs have been identified in Tribolium castaneum; however, none of the 41 GSTs has been functionally characterized. Here, three delta-class GSTs, TcGSTd1, TcGSTd2 and TcGSTd3, of T. castaneum were successfully cloned and expressed in Escherichia coli. All of the studied GSTs catalysed the conjugation of reduced glutathione with 1-chloro-2,4-dinitrobenzene. Insecticide treatment showed that the expression levels of TcGSTd3 and TcGSTd2 were significantly increased after exposure to phoxim and lambda-cyhalothrin, whereas TcGSTd1 was slightly upregulated only in response to phoxim. A disc diffusion assay showed that overexpression of TcGSTD3, but not TcGSTD1 or TcGSTD2, in E. coli increased resistance to paraquat-induced oxidative stress. RNA interference knockdown of TcGSTd1 caused metamorphosis deficiencies and reduced fecundity by regulating insulin/target-of-rapamycin signalling pathway-mediated ecdysteroid biosynthesis, and knockdown of TcGSTd3 led to reduced fertility and a decreased hatch rate of the offspring, probably caused by the reduced antioxidative activity in the reproductive organs. These results indicate that TcGSTd3 and TcGSTd2 may play vital roles in cellular detoxification, whereas TcGSTd1 may play essential roles in normal development of T. castaneum. These delta-class GSTs in T. castaneum have obtained different functions during the evolution.
Topics: Amino Acid Sequence; Animals; Female; Glutathione Transferase; Inactivation, Metabolic; Insect Proteins; Larva; Male; Pupa; Tribolium
PubMed: 31999035
DOI: 10.1111/imb.12637 -
Insect Science Feb 2022The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but...
The sigma glutathione S-transferases (GSTSs) are a class of cytosolic glutathione S transferases (GSTs) that play important roles in antioxidant defense in insects, but the mechanisms by which GSTSs contribute to antioxidant activity remain unclear. Here, we isolated a GSTS (GSTS6) from Tribolium castaneum and explored its function. Homology and phylogenetic analysis revealed that TcGSTS6 shared high identity with other evolutionarily conserved GSTSs. The recombinant TcGSTS6 protein had strong activity toward cumene hydroperoxide and 4-hydroxynonenal but low activity toward the universal substrate 1-chloro-2,4-dinitrobenzene. Exposure to various types of oxidative stress, including heat, cold, UV and pathogenic microbes, significantly induced TcGSTs6 expression, which indicates that it is involved in antioxidant defense. Knockdown TcGSTs6 by using RNA interference (RNAi) caused reduced antioxidant capacity, which was accomplished by cooperating with other antioxidant genes. Moreover, treatment with various insecticides such as phoxim, lambda-cyhalothrin, dichlorvos and carbofuran revealed that TcGSTS6 plays an important role in insecticide detoxification. The RNAi results showed that TcGSTS6 is essential for embryogenesis in T. castaneum. Our study elucidates the mechanism by which a GSTS contributes to antioxidant activity and enhances our understanding of the functional diversity of GSTSs in insects.
Topics: Animals; Embryonic Development; Glutathione Transferase; Inactivation, Metabolic; Insecticides; Phylogeny; Tribolium
PubMed: 34048152
DOI: 10.1111/1744-7917.12930 -
Environmental Pollution (Barking, Essex... Aug 2021Various chemicals containing pesticides can induce adipogenesis and cause obesity. Organophosphorus pesticides have been used for pest control. Here, we investigated the...
Organophosphorus pesticides exert estrogen receptor agonistic effect determined using Organization for Economic Cooperation and Development PBTG455, and induce estrogen receptor-dependent adipogenesis of 3T3-L1 adipocytes.
Various chemicals containing pesticides can induce adipogenesis and cause obesity. Organophosphorus pesticides have been used for pest control. Here, we investigated the estrogen receptor (ER)-dependent adipogenesis-inducing effect of representative organophosphorus pesticides (OPs), diazinon, phoxim, terbufos and tolclofos-methyl in 3T3-L1 adipocytes. Four OPs exhibited ER agonistic effect, determined using the OECD Performance Based Test Guideline No. 455; in vitro ER stably transfected transactivation assay using ERα-HeLa-9903 cell line, through binding affinity to ERα. Additionally, they increased lipid droplet accumulation in a dose-dependent manner, which was suppressed by ICI182,780, a well-known ER antagonist. Four OPs treatment induced peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein alpha (C/EBPα), and perilipin expression. Furthermore, PPARγ, C/EBPα and perilipin expression was inhibited by co-treatment with ICI182,780. The increased mRNA expression of lipoprotein lipase and fatty acid synthase by four OPs was suppressed by co-treatment with ICI182,780. These results indicated that diazinon, phoxim, terbufos, and tolclofos-methyl might have adipogenesis-inducing effect mediated by interacting with ER.
Topics: 3T3-L1 Cells; Adipocytes; Adipogenesis; Animals; Estrogens; Mice; Organisation for Economic Co-Operation and Development; Pesticides; Receptors, Estrogen
PubMed: 33872936
DOI: 10.1016/j.envpol.2021.117090 -
The Analyst Feb 2024Edible chrysanthemum is widely cultivated and used as an important ingredient of medicine, tea and multifunctional food. During the planting of chrysanthemum, pesticides...
Edible chrysanthemum is widely cultivated and used as an important ingredient of medicine, tea and multifunctional food. During the planting of chrysanthemum, pesticides are extensively used for preventing plant diseases and insect pests. To ensure the food safety of edible chrysanthemum, rapid detection methods are urgently needed for on-site inspection. In this study, a graphene oxide/Au nanoparticle (GO/Au NP) cellulose substrate was prepared through layer-by-layer assembly of GO and Au NPs on a mixed cellulose ester membrane. Surface-enhanced Raman spectroscopy (SERS) detection of four types of organophosphorus and organosulfur pesticides was achieved by filtering the extracting solution through the substrate and analysing SERS spectra. Qualitative and semi-quantitative detection of fenthion, phoxim, isocarbophos and thiram was accomplished with the detection limits of 38.01, 8.13, 48.97 and 8.74 ng mL, respectively. A spiking experiment further demonstrated the feasibility of this method for rapid and on-site detection of mixed pesticides in chrysanthemum. This study provides a new approach for rapid detection of multiple hazardous substances in flowering and herbal plants.
Topics: Pesticide Residues; Gold; Metal Nanoparticles; Pesticides; Spectrum Analysis, Raman; Cellulose; Graphite
PubMed: 38259149
DOI: 10.1039/d3an02030d -
International Journal of Molecular... Jan 2020The existence of a temperature effect of insecticides frustrated the control of the green plant bug (Meyer-Dür). Previous studies mostly focused on the application of... (Comparative Study)
Comparative Study
The existence of a temperature effect of insecticides frustrated the control of the green plant bug (Meyer-Dür). Previous studies mostly focused on the application of insecticides, but the underlying mechanism remains incompletely understood. Here, we report a transcriptome profiling of treated by three kinds of temperature coefficient insecticides (TCIs) (positive TCI: imidacloprid, negative TCI: b-cypermethrin and non-effect TCI: phoxim) at 15 °C, 25 °C and 35 °C by using next- and third-generation RNA-Seq methods. A total of 34,739 transcripts were annotated from 277.74 Gb of clean data. There were more up-regulated transcripts than down-regulated transcripts in all three kinds of TCI treatments. Further Venn diagrams indicate the regulatory transcripts and regulatory modes were different at the three temperatures. The responses to imidacloprid involved more detox and stress response transcripts such as cytochrome P450 (CYP450), carboxylesterase (CarE) and catalase (CAT) at 35 °C, which was the case for beta-cypermethrin at 15 °C. UDP-glucuronyltransferase (UGT) and heat shock protein (HSP) transcripts were heavily involved, and thus deserve particular note in the temperature effect of insecticides. This high-confidence transcriptome atlas provides improved gene information for further study on the insecticide temperature effect related physiological and biochemical processes of .
Topics: Animals; Gene Expression Profiling; Gene Expression Regulation; Heteroptera; High-Throughput Nucleotide Sequencing; Insect Proteins; Insecticides; Molecular Sequence Annotation; Neonicotinoids; Nitro Compounds; Organothiophosphorus Compounds; Pyrethrins; Sequence Analysis, RNA; Temperature
PubMed: 31963875
DOI: 10.3390/ijms21020658 -
Ecotoxicology and Environmental Safety Nov 2023Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is one of the most destructive insect pests owned strong resistance to different insecticides. Indoxacarb as a...
DIMBOA-induced gene expression, activity profiles of detoxification enzymes, multi-resistance mechanisms, and increased resistance to indoxacarb in tobacco cutworm, Spodoptera litura (Fabricius).
Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is one of the most destructive insect pests owned strong resistance to different insecticides. Indoxacarb as a novel oxadiazine insecticide becomes the main pesticide against S. litura. DIMBOA [2,4-dihydroxy-7-methoxy-2 H-1,4-benz-oxazin-3(4 H)-one] is involved in important chemical defense processes in corn plants. However, the insects' adaptation mechanism to insecticides when exposed to defensive allelochemicals in their host plants remains unclear. Here, we assessed multi-resistance, and resistance mechanisms based on S. litura life history traits. After 18 generations of selection, indoxacarb resistance was increased by 61.95-fold (Ind-Sel) and 86.06-fold (Dim-Sel) as compared to the Lab-Sus. Also, DIMBOA-pretreated larvae developed high resistance to beta-cypermethrin, chlorpyrifos, phoxim, chlorantraniliprole, and emamectin benzoate. Meanwhile, indoxacarb (LC) was applied to detect its impact on thirty-eight detoxification-related genes expression. The transcripts of SlituCOE073, SlituCOE009, SlituCOE074, and SlituCOE111 as well as SlGSTs5, SlGSTu1, and SlGSTe13 were considerably raised in the Ind-Sel strain. Among the twenty-three P450s, CYP6AE68, CYP321B1, CYP6B50, CYP9A39, CYP4L10, and CYP4S9v1 transcripts denoted significantly higher levels in the Ind-Sel strain, suggesting that CarEs, GSTs and P450s genes may be engaged in indoxacarb resistance. These outcomes further highlighted the importance of detoxification enzymes for S. litura gene expression and their role in responses to insecticides and pest management approaches.
Topics: Animals; Spodoptera; Insecticides; Nicotiana; Benzoxazines; Larva; Gene Expression; Insecticide Resistance
PubMed: 37944464
DOI: 10.1016/j.ecoenv.2023.115669 -
Pesticide Biochemistry and Physiology Jan 2021Dermanyssus gallinae poses a significant threat to poultry production, and the resistance to pyrethroids has been identified worldwide. Periodic monitoring of acaricide...
Dermanyssus gallinae poses a significant threat to poultry production, and the resistance to pyrethroids has been identified worldwide. Periodic monitoring of acaricide resistance in D. gallinae is very important for its control, and molecular mechanism associated with beta-cypermethrin resistance in D. gallinae is not fully clear. Results showed, four field isolates of CBP-1, CBP-2, CBP-5 and CBY-1 from China remained either susceptible or with decreased susceptibility (resistance ratio < 5.0) to phoxim, amitraz, propoxur and carbaryl. Four field isolates of CBP-1, CBP-3, CBY-2 and CBH-1 had developed high or extremely high level of resistance (resistance ratio ≥ 40.0) to beta-cypermethrin or permethrin. Detoxification enzyme activity of GSTs was significantly higher in beta-cypermethrin resistant (RS) than susceptible strain (SS), indicating that GSTs are probably involved in beta-cypermethrin resistance in D. gallinae. The recombinant GSTs (rGST-1, 2, 3) showed a pronounced activity toward the conjugates of 1-chloro-2, 4 dinitrobenzene (CDNB) and glutathione (GSH), with rGST-1 presenting the highest enzymatic activity. Constitutive over-expression of Deg-GST-2 was detected in RS strain, and GSTs genes were all inducible with the treatment of beta-cypermethrin in SS and RS strains. More importantly, knocking down Deg-GST-2 gene expression by RNAi increased the susceptibility of RS strain to beta-cypermethrin. HPLC analysis indicated that rGST-1 protein could metabolize phoxim directly, but rGSTs could not directly metabolize beta-cypermethrin. Our results indicated that some field isolates of D. gallinae from China had developed high level of resistance to pyrethroids, and elevated GSTs activity as well as increased GSTs expression levels were involved in beta-cypermethrin resistance, but the three evaluated GSTs did not play a direct role in the metabolism of beta-cypermethrin.
Topics: Acaricides; Animals; China; Glutathione; Mites; Pyrethrins; Transferases
PubMed: 33357546
DOI: 10.1016/j.pestbp.2020.104724 -
Food Chemistry Oct 2021Organophosphorus pesticides (OPPs) are widely used worldwide, leading to varying degrees of residues in food. Lactic acid bacteria (LAB) can degrade OPPs by producing...
Organophosphorus pesticides (OPPs) are widely used worldwide, leading to varying degrees of residues in food. Lactic acid bacteria (LAB) can degrade OPPs by producing phosphatase. This study explored the reasons for the variation in the degradation of different OPPs by Lactobacillus plantarum. The results showed that the degradation effects of OPPs by L. plantarum (intact cells) varied greatly, the degradation rate constant of phoxim was 1.65-fold higher than that of dichlorvos. However, the phosphatase extracted from L. plantarum had no degradation selectivity for OPPs in vitro. It was speculated that the selective uptake of cells determines this degradation selectivity. The results of molecular docking supported this hypothesis because there was no difference in the binding energies between phosphatase and OPPs, while the binding energies between phosphate-binding protein and pesticides were different, and they were negatively correlated with the degradation rate constants of the eight OPPs by L. plantarum.
Topics: Binding Sites; Chromatography, Gas; Kinetics; Lactobacillus plantarum; Molecular Docking Simulation; Organophosphorus Compounds; Organothiophosphorus Compounds; Pesticides; Phosphoric Monoester Hydrolases
PubMed: 34034058
DOI: 10.1016/j.foodchem.2021.130106 -
Spectrochimica Acta. Part A, Molecular... Aug 2024Based on the fact that not all chemical substances possess good Raman signals, this article focuses on the Raman silent region signals of pesticides with cyano group....
Based on the fact that not all chemical substances possess good Raman signals, this article focuses on the Raman silent region signals of pesticides with cyano group. Under the optimized conditions of methanol-water (1:1, v/v) as the solvent, irradiation at 302 nm light source for 20 min, and the use of 0.5 mol/L KI as the aggregating agent, Surface-enhanced Raman spectroscopy (SERS) method for azoxystrobin detection was developed by the Raman silent region signal of 2230 cm, and verified by detecting the spiked grapes with different concentrations of azoxystrobin. Other four pesticides with cyano group also could be identified at the peak of 2180 cm, 2205 cm, 2125 cm, and 2130 cm for acetamiprid, phoxim, thiacloprid and cymoxanil, respectively. When azoxystrobin or acetamiprid was mixed respectively with chlorpyrifos without cyano group, their SERS signals in the Raman silent region of chlorpyrifos were not interfered, while mixed with cymoxanil in different ratios (1:4, 1:1 and 4:1), respectively, each two pesticides with cyano group could be distinguished by the changes in the Raman silent region. In further, four pesticides with or without cyano group were mixed together in 1:1:1:1 (acetamiprid, cymoxanil, azoxystrobin chlorpyrifos), and each pesticide still could be identified even at 0.5 mg/L. The results showed that the SERS method combined with UV irradiation may provide a new way to monitor the pesticides with C≡N performance in the Raman silent region without interference from the food matrix.
Topics: Spectrum Analysis, Raman; Pesticides; Strobilurins; Pyrimidines; Vitis; Methacrylates; Neonicotinoids
PubMed: 38669978
DOI: 10.1016/j.saa.2024.124326