BACKGROUND: The Asian citrus psyllid, Diaphorina citri, is one of the major pests in citrus-growing areas around the world. The application of insecticides is the most effective method to reduce the population of D. citri. However, D. citri has developed resistance to multiple classes of insecticides. Understanding resistance mechanisms is crucial to the management of D. citri. In this study, molecular assays were performed to characterize imidacloprid resistance mechanisms. RESULTS: Based on the D. citri transcriptome database and other known insect resistance genes, 16 cytochrome P450, eight glutathione-S-transferase and six esterase genes were selected for cloning and sequencing. The gene expression analysis of 30 detoxification genes demonstrated that the relative expression of CYP4g15, CYP303A1, CYP4C62, CYP6BD5, GSTS1 and EST-6 were moderately high (>5-fold increase) in the imidacloprid-resistant strain. Feeding of double-stranded RNA (dsRNA) reduced the expression of the six genes (46.7%-72.1%) and resulted in significant adult mortality (65.62%-82.76%). We also determined the ability of different insecticides to induce the six selected genes. The expression of CYP4C62 and GSTS1 genes were the most significantly upregulated in adults treated with all insecticides, except for chlorfenapyr. In chlorfenapyr-treated D. citri, expression of CYP4g15 and CYP303A1 were the most highly induced. CONCLUSION: Overexpressed detoxification genes were associated with imidacloprid resistance, as confirmed by RNA interference feeding tests. The induction of the six selected genes when exposed to different insecticides supported the hypothesis that they were involved in the metabolism of the tested insecticides.
BACKGROUND: The Asian citrus psyllid, Diaphorina citri, is one of the major pests in citrus-growing areas around the world. The application of insecticides is the most effective method to reduce the population of D. citri. However, D. citri has developed resistance to multiple classes of insecticides. Understanding resistance mechanisms is crucial to the management of D. citri. In this study, molecular assays were performed to characterize imidacloprid resistance mechanisms. RESULTS: Based on the D. citri transcriptome database and other known insect resistance genes, 16 cytochrome P450, eight glutathione-S-transferase and six esterase genes were selected for cloning and sequencing. The gene expression analysis of 30 detoxification genes demonstrated that the relative expression of CYP4g15, CYP303A1, CYP4C62, CYP6BD5, GSTS1 and EST-6 were moderately high (>5-fold increase) in the imidacloprid-resistant strain. Feeding of double-stranded RNA (dsRNA) reduced the expression of the six genes (46.7%-72.1%) and resulted in significant adult mortality (65.62%-82.76%). We also determined the ability of different insecticides to induce the six selected genes. The expression of CYP4C62 and GSTS1 genes were the most significantly upregulated in adults treated with all insecticides, except for chlorfenapyr. In chlorfenapyr-treated D. citri, expression of CYP4g15 and CYP303A1 were the most highly induced. CONCLUSION: Overexpressed detoxification genes were associated with imidacloprid resistance, as confirmed by RNA interference feeding tests. The induction of the six selected genes when exposed to different insecticides supported the hypothesis that they were involved in the metabolism of the tested insecticides.