Contribution of cytochrome P450 monooxygenase CYP380C6 to spirotetramat resistance in Aphis gossypii Glover.

The cytochrome P450 monooxygenases play a key role in detoxification mechanism for spirotetramat resistance in Aphis gossypii Glover. However, only one P450 genes (CYP6DA2), among thirty-five P450 genes identified from Aphis gossypii transcriptome database, has been reported to play important role in spirotetramat resistance in previous resistance level until now. In this study, after the confirmation of the rise of resistance level and important roles of P450s in spirotetramat resistance by the synergism analysis, the gene expression changes were determined for P450 genes in spirotetramat susceptible and resistant strains. Compared with the susceptible strain, CYP6CY4, CYP6CY14, CYP6CY18 and CYP6DC1 in CYP3 Clade were up-regulated in resistant nymphs, with the CYP6CY14, CYP6CY4, CYP6DC1, and CYP6CY18 increased to 2.54-, 1.51-, 1.31- and 1.29-fold, respectively. Eight genes in CYP3 Clade, three genes in CYP4 Clade and one gene in Mito Clade were down-regulated. In resistant adult aphids, CYP380C6 in CYP4 Clade, CYP353B1 in CYP2 Clade, and CYP307A1 in Mito Clade were up-regulated under spirotetramat stress, with the CYP380C6, CYP353B1 and CYP307A1 increased to 2.89-, 1.91-, and 1.38-fold, respectively. In contrast, the other P450 genes were almost down-regulated, especially these P450 genes in CYP3 Clade, CYP4 Clade and Mito Clade. RNA interference of CYP380C6 significantly increased the sensitivity of the resistant adults and nymphs to spirotetramat, while suppression of CYP6CY14 could not increase the toxicity of spirotetramat. These results indicate the possible involvement of the CYP380C6 genes in spirotetramat resistance at present very high resistance levels. Screening the expression changes of P450 genes under different spirotetramat resistance levels in the genome-scale will provide an overall view on the possible metabolic factors in the resistance development. The results may facilitate further work to validate the roles of P450 in spirotetramat resistance with heterologous expression.