Identification and characterization of glutathione S-transferases and their potential roles in detoxification of abamectin in the rice stem borer, Chilo suppressalis.

The glutathione S-transferases (GSTs) are a kind of metabolic enzymes and participate in the detoxification metabolism of xenobiotics in various organisms. In insects, GSTs play important roles in the development of insecticide resistance and antioxidant protection. The rice stem borer Chilo suppressalis is one of the most damaging pests in rice and has developed high levels of resistance to abamectin in many areas of China, whereas the potential resistance mechanisms of C suppressalis to abamectin are still unclear. In the present study, a total of 23 CsGSTs genes were identified from the C. suppressalis transcriptome and genome, including 21 cytosolic and two microsomal CsGSTs. The cytosolic CsGSTs were further classified into seven categories based on phylogenetic analysis, and their sequence characteristics and genome structures were also analyzed. Synergism study revealed that the susceptibility of C. suppressalis to abamectin was increased significantly when the CsGSTs were inhibited by diethyl maleate (DEM). Sixteen CsGSTs genes were up-regulated in C. suppressalis larvae after treatment with abamectin, among which four CsGSTs genes including CsGSTe2, CsGSTe4, CsGSTo4 and CsGSTu1 were significantly induced in the midgut and fat body tissues. These results indicated that CsGSTs were associated with the detoxification of C. suppressalis to abamectin, and CsGSTe2, CsGSTe4, CsGSTo4 and CsGSTu1 might play important roles in the insecticide detoxification or antioxidant protection in C. suppressalis. Our present study provides valuable information on C. suppressalis GSTs, and are helpful in understanding the contributions of GSTs in abamectin detoxification in C. suppressalis and other insects.

Identification and Validation of ATP-Binding Cassette Transporters Involved in the Detoxification of Abamectin in Rice Stem Borer, Chilo suppressalis.

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.

Identification and transcriptional response of ATP-binding cassette transporters to chlorantraniliprole in the rice striped stem borer, Chilo suppressalis.

BACKGROUND: As the largest transporter gene family in metazoans, ATP-binding cassette (ABC) transporters regulate the efflux of a broad spectrum of substrates from the cytoplasm to the outside of the cell. In arthropods, ABC transporters are involved in phase III of the detoxification process, and play important roles in the metabolism and transport of insecticides. RESULTS: We identified 54 ABC transporters from the genome and transcriptome of Chilo suppressalis, one of the most damaging pests of rice in China. The identified ABC transporters were classified into eight subfamilies (ABCA to ABCH) based on NCBI BLAST and phylogenetic analysis. Synergism studies showed that treatment with verapamil, a potent inhibitor of ABC transporters, resulted in significantly increased toxicity of chlorantraniliprole against C. suppressalis larvae. Among the 21 tested ABC genes, three ABC transporter genes including CsABCC8, CsABCG1C and CsABCH1 were significantly upregulated after chlorantraniliprole treatment. CONCLUSION: ABC transporters play important roles in the detoxification and transport of chlorantraniliprole in C. suppressalis. The results from our study provide valuable information on C. suppressalis ABC transporters, and are helpful in understanding the roles of ABC transporters in chlorantraniliprole resistance mechanisms in C. suppressalis and other insect pests. © 2020 Society of Chemical Industry.