No abstract

The mammalian heme oxygenase (HO) plays an important role in cytoprotection against oxidative-stress-induced cell damage; however, functional characterization of insect HO is still limited. In this study, cDNA encoding a HO, named Sf HO, was cloned from Spodoptera frugiperda. Analysis of the transcription level and enzymatic activity showed that exposure of the LC 30 concentration of chlorantraniliprole to the third instar larvae significantly upregulated both the mRNA level and enzymatic activity of Sf HO at 24 h after treatment. Further injection of the HO activator, hemin, into the third instar larvae led to the upregulation of Sf HO as well as decreased susceptibility of S. frugiperda to chlorantraniliprole. Consistently, overexpression of Sf HO increased the Sf9 cell viability under chlorantraniliprole treatment. Strikingly, both RNAi and the dual-luciferase reporter assay in Sf9 cells revealed that, unlike mammalian HO that is regulated by the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), Sf HO was not subject to the regulation by cap 'n' collar isoform C (CncC), the Nrf2 homologue in insects. These data provide insights into the function and regulatory mechanism of insect HOs and had applied implications for the control of S. frugiperda.

Chilo suppressalis has developed high levels of resistance to abamectin in many areas of China, while the underline resistance mechanisms are largely unclear. ATP-binding cassette (ABC) transporters function in transporting a large diversity of substrates including insecticides and play important roles in the detoxification metabolism of insects. In this study, synergism bioassay revealed that the ABC transporters were involved in the detoxification of C. suppressalis to abamectin. Six ABC transporter genes were upregulated in C. suppressalis after abamectin exposure, among which five genes CsABCC8, CsABCE1, CsABCF1, CsABCF2, and CsABCH1 were induced in the detoxification-related tissues. In addition, the five ABC transporters were recombinantly expressed in Sf9 cells, and the cytotoxicity assay showed that the viabilities of cells expressing CsABCC8 or CsABCH1 were significantly increased when compared with the viabilities of cells expressing EGFP after abamectin, chlorantraniliprole, cyantraniliprole, fipronil, and chlorpyrifos treatment, respectively. Overexpression of CsABCE1 significantly increased the viabilities of cells to abamectin, chlorantraniliprole, deltamethrin, and indoxacarb exposure, respectively. These results suggested that CsABCC8, CsABCE1, and CsABCH1 might participate in the detoxification and transport of abamectin and several other classes of insecticides in C. suppressalis. Our study provides valuable insights into the transport-related detoxification mechanisms in C. suppressalis and other insects.

The glutamate-gated chloride channels (GluCls) play essential roles in signal transduction by regulating fast inhibitory synaptic transmission in the nervous system of invertebrates. While there is only one GluCl subunit in the insect, the diversity of insect GluCls is broadened by alternative splicing. In the present study, three TcGluCl variant genes were cloned from the red flour beetle Tribolium castaneum. Analysis of the characteristics of TcGluCls including sequence features, genomic structures, and alternative splicing revealed that TcGluCls had the typical structural features of GluCls and showed high homologies with the GluCls from other insect orders. The TcGluCl-encoding gene consists of nine exons and three variants (TcGluCl-3a, TcGluCl-3b, and TcGluCl-3c) were generated by the alternative splicing of exon 3, which was a highly conserved alternative splicing site in insect GluCls. Homology modeling of TcGluCl-3a showed that the exon 3 coding protein located at the N-terminal extracellular domain, and there were no steric clashes encountered between the exon 3 coding region and ivermectin/glutamate binding pocket, which indicated that the alternative splicing of exon 3 might have no impact on the binding of GluCls to glutamate and insecticide. In addition to the head tissue, TcGluCl-3a and TcGluCl-3c also had high expressions in the ovary and testis of T. castaneum, whereas TcGluCl-3b showed high expression in the midgut, suggesting the diverse physiological functions of TcGluCl variants in T. castaneum. The total TcGluCl and three variants showed the highest expression levels in the early stage larvae. The expressions of TcGluCl, TcGluCl-3b, and TcGluCl-3c were significantly increased from the late-stage larvae to the early stage pupae and indicated that the TcGluCl might be involved in the growth and development of T. castaneum. These results are helpful to further understand the molecular characteristics of insect GluCls and provide foundations for studying the specific function of the GluCl variant.

No abstract

BACKGROUND The selective insecticide flonicamid shows highly insecticidal activities against piercing‐sucking insects and has been widely used for the control of Hemipteran insect pests, whereas its effects on Lepidopteran insect pests remain largely unknown. Recently, inward rectifier potassium (Kir) channel has been verified to be a target of flonicamid, however, functional characterization of Lepidopteran Kir genes is still lacking. RESULTS Flonicamid shows no insecticidal toxicity against Chilo suppressalis larvae. However, the feeding and growth of larvae were reversibly inhibited by flonicamid (50–1200 mg L−1). Flonicamid treatment also remarkably reduced and delayed the pupation and eclosion of Chilo suppressalis. Additionally, five distinct Kir channel genes (CsKir1, CsKir2A, CsKir2B, CsKir3A and CsKir3B) were cloned from Chilo suppressalis. Expression profiles analysis revealed that CsKir2A was predominately expressed in the hindgut of larvae, whereas CsKir2B had high expressions in the Malpighian tubules and hindgut. RNA interference (RNAi)‐mediated knockdown of CsKir2B significantly reduced the growth and increased the mortalities of larvae, whereas silencing of CsKir2A had no obvious effects on Chilo suppressalis. CONCLUSION Flonicamid exhibits adverse effects on the growth and development of Chilo suppressalis. CsKir2B might be involved in the feeding behavior of Chilo suppressalis. These results provide valuable information on the effects of flonicamid on non‐target insects as well as the function of insect Kir channels, and are helpful in developing new insecticide targeting insect Kir channels. © 2020 Society of Chemical Industry

BACKGROUND Sublethal exposure to insecticides causes changes in insect behaviors and physiologies including feeding, mobility, communication, hormone homeostasis, development and fecundity, however, the underlying molecular mechanisms were largely unclear. Our previous studies revealed that sublethal chlorantraniliprole exposure disturbed the hormone homeostasis, reduced the weight and longevity and prolonged the developmental duration of Chilo suppressalis. In the present study, the potential phosphorylation modification regulation mechanisms in C. suppressalis in response to sublethal chlorantraniliprole exposure were explored using comparative and quantitative phosphoproteomics. RESULTS A total of 2640 phosphopeptides belonging to 1144 phosphoproteins were identified, among which 446 phosphopeptides derived from 303 unique phosphoproteins were differentially phosphorylated between the chlorantraniliprole‐treated and control larvae. The phosphorylation levels of differentially phosphorylated phosphopeptides were further validated using parallel reaction monitoring (PRM). Functional classification and protein–protein interaction of the differentially phosphorylated proteins (DPPs) were analyzed. Generalized analysis of the DPPs and the differentially expressed genes (DEGs) identified in our previous study showed that sublethal chlorantraniliprole exposure significantly changed the transcription and phosphorylation levels of genes/proteins associated with carbohydrate and lipid metabolism, cytoskeleton, signal transduction, transcription, translation and post‐translational modification, leading to the dysfunctions of energy metabolism, transcription regulation, protein synthesis and modification, and signal transduction in C. suppressalis. Further analysis of the phosphorylation motifs in DPPs revealed that the MAPKs, CDKs, CaMK II, PKA, PKC and CK II protein kinases might be directly responsible for the phosphoproteomics response of C. suppressalis to chlorantraniliprole treatment. CONCLUSION Our results provide abundant phosphorylation information for characterizing the protein modification in insects, and also provide valuable insights into the molecular mechanisms of insect post‐translational modifications in response to sublethal insecticide exposure. © 2023 Society of Chemical Industry.