PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2025
·
IF 4
Yu-Xi Liu,Chao Hu,Zi-Nan Xia,Yan Wang,Yu-Ting Li,Ping Gao,Yun-Tong Lv,Jia Li,Xue-Qing Yang
G protein-coupled receptors (GPCRs) constitute a diverse and crucial family of membrane receptors, regulating a wide array of physiological processes. Although the involvement of GPCR signaling pathways in modulating key genes associated with insecticide resistance has been documented in various insect species, the molecular mechanisms underlying GPCR-mediated resistance in Cydia pomonella remain largely unknown. To elucidate the molecular basis of lambda-cyhalothrin (LCT) resistance in C. pomonella , we performed comparative transcriptome sequencing on whole-body, head, and midgut tissues of larvae from a susceptible population (SS) and a laboratory-selected LCT-resistant population (LCR) derived from SS. Five GPCR candidates ( CpGPCRs ) exhibiting overexpression in LCR were identified, including two members of Family A ( CpGPCR4 and CpGPCR55 ), two of Family B ( CpGPCR68 and CpGPCR76 ), and one of Family F ( CpGPCR94 ). Spatiotemporal expression profiling revealed that these genes were predominantly expressed during the first-instar larval and pupal-adult stages, with notably high expression levels in the larval heads. Notably, all genes exhibited significantly elevated expression in the midgut of LCR larvae compared to SS larvae. RNA interference (RNAi)-mediated knockdown of CpGPCR4 , CpGPCR55 , and CpGPCR68 increased larval susceptibility to LCT, while knockdown of CpGPCR4 and CpGPCR55 significantly reduced larval survival rates. Furthermore, larvae treated with dsGPCR55 exhibited molting defects and a decline in 20-hydroxyecdysone (20E) titers. These findings demonstrate that CpGPCRs play essential roles in the survival and development of C. pomonella and are critically involved in mediating resistance to LCT. This study enhances our understanding of the molecular mechanisms driving insecticide resistance and identifies potential targets for the development of novel pest management strategies against C. pomonella .
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BULLETIN OF ENTOMOLOGICAL RESEARCH
·
2015
·Vol. 105
·期 3
·316-325
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IF 1.91
X.-Q. Yang,Y.-L. Zhang
The codling moth Cydia pomonella (L.) is an economically important fruit pest and it has been directly targeted by insecticides worldwide. Serious resistance to insecticides has been reported in many countries. As one of the most serious invasive pest, the codling moth has populated several areas in China. However, resistance to insecticides has not been reported in China. We investigated the insecticide-resistance status of four field populations from Northwestern China by applying bioassays, enzyme activities, and mutation detections. Diagnostic concentrations of lambda-cyhalothrin, chlorpyrifos-ethyl, carbaryl, and imidacloprid were determined and used in bioassays. Field populations were less susceptible to chlorpyrifos-ethyl and carbaryl than laboratory strain. Insensitive populations displayed an elevated glutathione S-transferases (GSTs) activity. Reduced carboxylesterase (CarE) activity was observed in some insecticide insensitive populations and reduced acetylcholinesterase activity was observed only in the Wuw population. The cytochrome P450 polysubstrate monooxygenases activities in four field populations were not found to be different from susceptible strains. Neither the known-resistance mutation F399V in the acetylcholinesterase (AChE) gene, ace1, nor mutations in CarE gene CpCE-1 were found in adult individuals from our field populations. Native-PAGE revealed that various CarE isozymes and AChE insensitivity were occurring among Chinese populations. Our results indicate that codling moth populations from Northwestern China were insensitivity to chlorpyrifos-ethyl and carbaryl. Increased GST activity was responsible for insecticides insensitivity. Decreased CarE activity, as well as the presence of CarE and AChE polymorphisms might also be involved in insecticides insensitivity. New management strategies for managing this pest are discussed.
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CABI Agriculture & Bioscience
·
2023
·Vol. 4
·期 1
·1-17
·
IF 3.9
Ju Di,Hu Chao,Li Peirong,Gao Ping,Li Yuting,Yang Xueqing
Background The primary method utilized by orchard owners to combat Cydia pomonella is the application of various chemical insecticides. However, this has resulted in the development of resistance. The resistance mechanisms to insecticides from different chemical classes are diverse but interconnected. Therefore, it is crucial to comprehend the commonalities in these mechanisms to effectively develop strategies for managing resistance.Materials and methodsTo determine whether target-site insensitivity to LCT and AM plays a role in resistance, the sequences of voltage-gated sodium channel (VGSC) and glutamate-gated chloride channel (GluCl) containing the mutation domains were detected. To validate whether similar mechanisms were involved in the detoxification process of lambda-cyhalothrin (LCT) and abamectin (AM) at sublethal doses (specifically LD10 and LD30), cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST), and carboxylesterases (CarE) activities were evaluated after insecticides exposure; synergistic experiments were conducted using piperonyl butoxide (PBO), diethyl maleate (DEM), and triphenyl phosphate (TPP) as inhibitors of P450, GST, and CarE respectively. RNA-sequencing (RNA-Seq) was performed to compare the expression levels of detoxification-related genes between susceptible (SS) and resistant strains.Results The best known target-site mutations caused by LCT and AM, including L1014F in VGSC and V263I, A309V, I321T, and G326E in GluCl were not occurred. We observed that PBO had a strong synergistic effect on LCT and AM, while DEM on LCT. The activities of detoxification enzymes increased after insecticide exposures, indicating that the detoxification of LCT was primarily carried out by P450 and CarE enzymes, while P450 and GST enzymes played a major role in the detoxification of AM. A total of 72 P450 genes and 75 CarE genes were identified in the C. pomonella transcriptome, with 43 of these genes (including 11 P450, 3 GST, 10 CarE, 11 ABC transporters, and 8 UDP-glycosyl transferases) being over-expressed in response to both insecticides Interestingly, ABC transporters were predominantly induced by AM treatment, while GST showed higher induction levels with LCT treatment. Furthermore, LCT-resistant strains of C. pomonella exhibited higher levels of induction of detoxification-related genes compared to susceptible strains.Conclusion The up-regulation of these detoxification genes is a common metabolic mechanism employed by C. pomonella to counteract the effects of insecticides, although the extent of gene expression change varies depending on the specific insecticide.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2019
·
IF 3.571
Wei Wang,Chao Hu,Xin-Ru Li,Xiao-Qi Wang,Xue-Qing Yang
Little is known about the role of specific delta GST genes in the detoxification of lambda-cyhalothrin in the global quarantine fruit pest codling moth, Cydia pomonella (L.). Real-time quantitative PCR shows that CpGSTd3 was ubiquitously expressed at all developmental stages and is most abundant in the larval stage and lowest in the egg stage; the mRNA level of CpGSTd3 is higher in the midgut and Malpighian tubules of fourth-instar larvae and abdomens of adults than in other tissues. Exposure of fourth-instar larvae to an LD10 dosage of lambda-cyhalothrin significantly induced the transcript of CpGSTd3 at 3 h, but the mRNA level was down-regulated after 12 h of treatment. Recombinant CpGSTd3 expressed in Escherichia coli was able to catalyze the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and with an IC50 value of 0.65 mM for lambda-cyhalothrin. Metabolism assays indicate that recombinant CpGSTd3 could metabolize lambda-cyhalothrin. These results suggest that CpGSTd3 is probably a lambda-cyhalothrin metabolizing GST in C. pomonella.
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INSECT MOLECULAR BIOLOGY
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2016
·Vol. 25
·期 6
·769-777
·
IF 2.866
Z. Tian,Y. Zhang
A full-length cDNA encoding Cydia pomonella pheromone binding protein 1 (CpomPBP1) was cloned and characterized. CpomPBP1, possessing the typical characteristics of lepidopteran odorant binding proteins, was detected to be specifically expressed in the antennae of male and female moths at the mRNA and protein level. Soluble recombinant CpomPBP1 was subjected to in vitro binding to analyse its binding properties and to search for potentially active semiochemicals. A competitive binding assay showed that three 12-carbon ligands, codlemone, 1-dodecanol and E,E-2,4-dodecadienal, were able to bind to CpomPBP1 in decreasing order of affinity. Moreover, unlike the wild-type CpomPBP1, the C-terminus truncated CpomPBP1 exhibited high affinity to ligands even in an acidic environment, suggesting that the C-terminus plays a role in preventing ligands from binding to CpomPBP1 in a lower pH environment.
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2014
·Vol. 98
·期 21
·8947-8962
·
IF 3.811
Liu Jiyuan,Yang Xueqing,Zhang Yalin
In insects, glutathione S -transferases (GSTs) are enzymes involved in detoxification of insecticides. However, few data are available for the codling moth, Cydia pomonella (L.). In this study, we cloned a delta class GST gene CpGSTd1 from C. pomonella . Real-time quantitative PCR shows that CpGSTd1 was up-regulated with aging, and the mRNA level of CpGSTd1 was higher in the fat body and silk glands than in other tissues. The expression level of CpGSTd1 exposure to insecticide suggests that CpGSTd1 is up-regulated after chlorpyrifos-methyl and lambda-cyhalothrin treatments. Both lambda-cyhalothrin and chlorpyrifos-methyl altered GST activity in vivo. The purified CpGSTd1 protein exhibits a high catalytic efficiency with CDNB and was inhibited by lambda-cyhalothrin and chlorpyrifos-methyl in vitro. Metabolism assays indicate that lambda-cyhalothrin was significantly metabolized while chlorpyrifos-methyl was not metabolized by CpGSTd1. Binding free energy analysis suggests that CpGSTd1 binding is tighter with lambda-cyhalothrin than with chlorpyrifos-methyl. Our study suggests that CpGSTd1 plays a key role in the metabolism of insecticides in C. pomonella .
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NEOTROPICAL ENTOMOLOGY
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2026
·Vol. 55
·期 1
·9
·
IF 1.8
Zheng Kang-Wu,Wei Zi-Han,Xia Zi-Nan,Wang Bo-Kun,Gao Ping,Li Yu-Ting,Yang Xue-Qing
Rising ambient temperatures driven by global warming are subjecting insect populations to thermal conditions exceeding their historical environmental ranges. Understanding how temperature alters key life-history traits is essential for optimizing integrated pest management strategies, particularly for invasive species. This study investigated the effects of short-term thermal stress (31 °C for the parental generation, with offspring returned to 26℃) on life history traits and insecticide sensitivity in the codling moth Cydia pomonella (L.), a major agricultural invasive pest. Results demonstrated that elevated temperature significantly shortened larval developmental duration and reduced pupal weight and overall fitness, with these phenotypic effects persisting across generations. Additionally, increased temperature attenuated the sensitivity of C. pomonella to lambda -cyhalothrin, especially in parental generation. In contrast, reproductive output and sensitivity to chlorantraniliprole remained largely unaffected by thermal elevation. Activities of detoxification enzymes including cytochrome P450 monooxygenases (P450s), carboxylesterases (CarEs), and the antioxidant enzyme catalase (CAT), were significantly upregulated at elevated temperatures, with transgenerational inheritance observed in progeny, potentially contributing to the observed decrease in insecticide sensitivity. These findings advance our understanding of the transgenerational physiological and biochemical adaptions of C. pomonella to thermal stress, with implications for resistance management under climate change scenarios.
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2026
·
IF 4.8
Yu-Ze Li,Chao Hu,Yu-Xi Liu,Yun-Tong Lv,Xue-Qing Yang
Extensive studies have demonstrated that cytochrome P450 enzymes play a critical role in the metabolic detoxification and resistance development of pests against insecticides; however, relative to P450 genes in Clan 3, knowledge on mitochondrial Clan members remains limited. Here, we report the involvement of a CYP333 family member, CYP333B119v2 , in conferring lambda -cyhalothrin resistance in Cydia pomonella . Phylogenetic analysis shows that CYP333B119v2 clusters with CYP333 family members of Lepidoptera, it containing three highly conserved P450 domains: the Heme-binding region, C-Helix, and K-helix. Spatiotemporal expression profiling indicates that CYP333B119v2 transcription levels increase progressively during larval development, peaking at the fourth instar, with significant expression in the midgut, the primary detoxification tissue. Relative to susceptible population (SS), CYP333B119v2 expression is 2.73-fold higher in lambda -cyhalothrin moderate resistant population (LCR) and is significantly induced by sublethal doses of lambda -cyhalothrin . Knockdown of CYP333B119v2 enhances larval susceptibility to lambda -cyhalothrin. Molecular docking based on AlphaFold models revealed strong binding interactions between CYP333B119v2 and lambda -cyhalothrin, with diverse non-covalent interactions involved in the binding process. These findings suggest that CYP333B119v2 may contribute to lambda -cyhalothrin resistance in C. pomonella . This study is the first report implicating a mitochondrial CYP333B subfamily member in the resistance mechanisms of Lepidoptera, advancing our understanding of P450-mediated metabolic resistance and providing new insights into pest management strategies.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2023
·
IF 6.1
Yu-Xi Liu,Chao Hu,Yu-Ting Li,Ping Gao,Xue-Qing Yang
While previous studies have reported G protein-coupled receptor (GPCR)-mediated insecticide resistance in various arthropods, the understanding of GPCR-associated resistance mechanisms in Cydia pomonella remains limited. In this study, a total of 95 CpGPCR genes categorized into four families were identified in C. pomonella. Results revealed high expression levels of the majority of the CpGPCRs during the first larval stage and in the head of C. pomonella. Exposure to lambda-cyhalothrin significantly increased the expression of 15 CpGPCRs, including CpGPCR70, which is highly expressed in all larval stages and shows the highest expression in the midgut. RNA interference (RNAi) demonstrated that downregulation of CpGPCR70 leads to reduced expression of key resistance-related genes and a decreased tolerance of larvae to lambda-cyhalothrin. These findings indicate that CpGPCR70 plays a crucial role in regulating the expression of detoxifying genes involved in lambda-cyhalothrin resistance, offering valuable insights for the development of more effective pest control strategies.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2023
·
IF 6.1
Chao Hu,Yu-Xi Liu,Shi-Pang Zhang,Ya-Qi Wang,Ping Gao,Yu-Ting Li,Xue-Qing Yang
Aryl hydrocarbon receptor (AhR) enhances insect resistance to insecticides by regulating the detoxification network. Our previous studies have confirmed that overexpressions of cytochrome P450 monooxygenases (P450s) and glutathione S-transferases (GSTs) are involved in lambda-cyhalothrin resistance in Cydia pomonella. Here, we report that CpAhR regulates the expression of GST and P450 genes, thus conferring resistance. Expression patterns indicated that the expression of CpAhR was highly induced by lambda-cyhalothrin exposure and upregulated in a lambda-cyhalothrin-resistant population. RNA interference (RNAi) of CpAhR decreases the expression of key resistance-related genes (CpGSTe3, CpCYP9A121, and CpCYP9A122) and the activity of the GST enzyme, reducing the tolerance to lambda-cyhalothrin. Furthermore, β-naphthoflavone, a novel agonist of AhR, was first proven to be effective in increasing CpAhR expression and larval tolerance to lambda-cyhalothrin. These results demonstrate that CpAhR regulates the expression of key detoxifying genes and GST activity, resulting in the development of resistance to lambda-cyhalothrin in C. pomonella.
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Scientific Reports
·
2016
·Vol. 6
·期 1
·1-11
·
IF 5.228
Tian Zhen,Liu Jiyuan,Zhang Yalin
Given the advantages of behavioral disruption application in pest control and the damage of Cydia pomonella , due progresses have not been made in searching active semiochemicals for codling moth. In this research, 31 candidate semiochemicals were ranked for their binding potential to Cydia pomonella pheromone binding protein 2 (CpomPBP2) by simulated docking and this sorted result was confirmed by competitive binding assay. This high predicting accuracy of virtual screening led to the construction of a rapid and viable method for semiochemicals searching. By reference to binding mode analyses, hydrogen bond and hydrophobic interaction were suggested to be two key factors in determining ligand affinity, so is the length of molecule chain. So it is concluded that semiochemicals of appropriate chain length with hydroxyl group or carbonyl group at one head tended to be favored by CpomPBP2. Residues involved in binding with each ligand were pointed out as well, which were verified by computational alanine scanning mutagenesis. Progress made in the present study helps establish an efficient method for predicting potentially active compounds and prepares for the application of high-throughput virtual screening in searching semiochemicals by taking insights into binding mode analyses.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Chao Hu,Cong Zhang,Yi-Fan Tang,Yu-Xi Liu,Zi-Nan Xia,Yan Wang,Wei-Tu Li,Ping Gao,Yu-Ting Li,Yun-Tong Lv,Xue-Qing Yang
Insecticides are commonly utilized in agriculture and forestry for pest control, but their dispersal can pose hazards to humans and environment. Understanding resistance, inheritance patterns, and fitness costs can help manage resistance. A λ-cyhalothrin-resistant population (LCR) of Cydia pomonella, a global pest of pome fruits and walnuts, was obtained through selective insecticide breeding for 15 generations, showing stable moderate resistance (23.85-fold). This population was cross-resistant to deltamethrin (4.26-fold) but not to β-cypermethrin, chlorantraniliprole, chlorpyrifos, and avermectin. Genetic analysis revealed the resistance was autosomal, incompletely dominant, and controlled by multiple genes. Increased activity of glutathione S-transferases and cytochrome P450 monooxygenases (P450s) played a primary role in resistance, with specific genes up-regulated in LCR, and exhibited significant expression in midgut. LCR also exhibited fitness costs, including delays in development, reduced fecundity, and slower population growth. These findings contribute to understanding λ-cyhalothrin resistance in C. pomonella and can guide resistance management strategies.
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