Cytochrome P450 and UDP-glucosyltransferase (UGT) as phase I and phase II metabolism enzymes, respectively, play vital roles in the breakdown of endobiotics and xenobiotics. Insects can increase the expression of detoxification enzymes to cope with the stress from xenobiotics including insecticides. However, the molecular mechanisms for insecticide detoxification in Spodoptera exigua remain elusive, and the genes conferring insecticide metabolisms in this species are less well reported. In this study, 68 P450 and 32 UGT genes were identified. Phylogenetic analysis showed gene expansions in CYP3 and CYP4 clans of P450 genes and UGT33 family of this pest. P450 and UGT genes exhibited specific tissue expression patterns. Insecticide treatments in fat body cells of S. exigua revealed that the expression levels of P450 and UGT genes were significantly influenced by challenges of abamectin, lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb. Multiple genes for detoxification were affected in expression levels after insecticide exposures. The results demonstrated that lambda-cyhalothrin, chlorantraniliprole, metaflumizone and indoxacarb induced similar responses in the expression of P450 and UGT genes in fat body cells; eight P450 genes and four UGT genes were co-up-regulated significantly, and no or only a few CYP/UGT genes were down-regulated significantly by these four insecticides. However, abamectin triggered a distinct response for P450 and UGT gene expression; more P450 and UGT genes were down-regulated by abamectin than by the other four compounds. In conclusion, P450 and UGT genes from S. exigua were identified, and different responses to abamectin suggest a different mechanism for insecticide detoxification.
The lethal and sublethal effects of chlorantraniliprole against Spodoptera exigua (Hübner) were evaluated under laboratory conditions by oral exposure of neonate larvae to the compound. The 72 h LC 50 value of this insecticide to S. exigua was found to be 12.747 lg l -1 . A progressive larval mortality of 24.32% for LC 30 treatment and 42.61% for LC 50 treatment was observed from 4th to 6th day after exposure, which resulted in the reduced pupation rates in exposure groups. The sublethal effects of this chemical were indicated by prolongation of larval period, the increase of pupal weight and decrease in hatch rate of egg. Chlorantraniliprole at LC 30 and LC 50 rate significantly delayed larval development; the developmental duration of surviving larvae was extended for 22.5 and 28.6%, respectively, compared with that of control group. LC 30 treatment increased the mean weight of pupa and induced to the production of heavier pupa ([150 mg). In LC 50 treatment, heavier pupa also showed up but the mean weight of pupa was not influenced. The egg hatch rate in LC 50 group was significant lower than that in control and LC 30 groups. No significant differences in pupal duration, emergence rate, sex ratio, egg number per female, and longevity of adults were observed among treatments. Chlorantraniliprole had exceptional activity against S. exigua according to concentration-response bioassay in laboratory, and the toxicities were primarily resulted from immediate lethality.
The evolution of insect resistance to insecticides is frequently associated with overexpression of one or more cytochrome P450 enzyme genes. Although overexpression of CYP450 genes is a well-known mechanism of insecticide resistance, the underlying regulatory mechanisms are poorly understood. Here we uncovered the mechanisms of overexpression of the P450 gene, CYP321A8 in a major pest insect, Spodoptera exigua that is resistant to multiple insecticides. CYP321A8 confers resistance to organophosphate (chlorpyrifos) and pyrethroid (cypermethrin and deltamethrin) insecticides in this insect. Constitutive upregulation of transcription factors CncC/Maf are partially responsible for upregulated expression of CYP321A8 in the resistant strain. Reporter gene assays and site-directed mutagenesis analyses demonstrated that CncC/Maf enhanced the expression of CYP321A8 by binding to specific sites in the promoter. Additional cis-regulatory elements resulting from a mutation in the CYP321A8 promoter in the resistant strain facilitates the binding of the orphan nuclear receptor, Knirps, and enhances the promoter activity. These results demonstrate that two independent mechanisms; overexpression of transcription factors and mutations in the promoter region resulting in a new cis-regulatory element that facilitates binding of the orphan nuclear receptor are involved in overexpression of CYP321A8 in insecticide-resistant S. exigua.
The whitebacked planthopper, Sogatella furcifera (Horváth), and small brown planthopper, Laodelphax striatellus (Fallén), both are important crop pests throughout China, especially in rice. Application of chemical insecticides is the major control practice. Consequently, insecticide resistance has become an urgent issue. In this study, resistance levels to six conventional insecticides were evaluated for these two species collected from major occurring areas of China. Additionally, imidacloprid- (resistance ratio [RR] = 10.4-fold) and buprofezin (RR = 15.1-fold)-resistant strains of whitebacked planthopper were obtained through laboratory selections for cross-resistance profiling and synergism assessment to understand resistance mechanisms. The results showed that all tested populations of both species exhibited low to high levels of resistance to chlorpyrifos, while remaining susceptible to thiamethoxam. Three of the 14 whitebacked planthopper populations showed low to moderate resistance to imidacloprid, while all small brown planthopper populations reminded susceptible. All small brown planthopper and whitebacked planthopper (except one) populations showed at least moderate resistance (RR = 10.1-271.1) to buprofezin. All small brown planthopper populations remained susceptible to pymetrozine and nitenpyram, and all whitebacked planthopper populations remained susceptible to isoprocarb. The imidacloprid-resistant whitebacked planthopper strain showed no significant cross-resistance to other tested insecticides. However, the buprofezin-resistant strain exhibited a low-level cross-resistance (CR = 3.1) to imidacloprid. Piperonyl butoxide, triphenyl phosphate, and diethylmaleate displayed no synergism effect on the resistant whitebacked planthopper strains.
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