The dynamic changes of genes revealed that persistently overexpressed genes drive the evolution of cyflumetofen resistance in Tetranychus cinnabarinus

Abstract

Changes in gene expression are associated with the evolution of pesticide resistance in arthropods. In this study, transcriptome sequencing was performed in 3 different resistance levels (low, L; medium, M; and high, H) of cyflumetofen-resistant strain (YN-CyR). A total of 1 685 genes, including 97 detoxification enzyme genes, were upregulated in all 3 stages, of which 192 genes, including 11 detoxification enzyme genes, showed a continuous increase in expression level with resistance development (L to H). RNA interference experiments showed that overexpression of 7 genes (CYP392A1, TcGSTd05, CCE06, CYP389A1, TcGSTz01, CCE59, and CYP389C2) is involved in the development of cyflumetofen resistance in Tetranychus cinnabarinus. The recombinant CYP392A1 can effectively metabolize cyflumetofen, while CCE06 can bind and sequester cyflumetofen in vitro. We compared 2 methods for rapid screening of resistance molecular markers, including short-term induction and 1-time high-dose selection. Two detoxification enzyme genes were upregulated in the field susceptible strain (YN-S) by induction with 20% lethal concentration (LC 20 ) of cyflumetofen. However, 16 detoxification enzyme genes were upregulated by 1-time selection with LC 80 of cyflumetofen. Interestingly, the 16 genes were overexpressed in all 3 resistance stages. These results indicated that 1 685 genes that were upregulated at the L stage constituted the basis of cyflumetofen resistance, of which 192 genes in which upregulation continued to increase were the main driving force for the development of resistance. Moreover, the 1-time high-dose selection is an efficient way to rapidly obtain the resistance-related genes that can aid in the development of resistance markers and resistance management in mites.

UGT201H1 overexpression confers cyflumetofen resistance in Tetranychus cinnabarinus (Boisduval)

“…7,52 In YN-CyR, transcriptome sequencing at different resistance levels revealed that the expression of P450, GST and CCE genes were all upregulated. 50 Interestingly, synergist assays suggest that the P450s are the key detoxification enzymes that mediate the formation of resistance to cyflumetofen in YN-CyR. 53 This is counter to the findings in CyR, and the reason for this difference in CyR and YN-CyR may be the different backgrounds of the initial strains (SS and YN-S, respectively).…”

“…Our previous study demonstrated that CyR (selected from SS by cyflumetofen in the laboratory) and YN-CyR strains were both polygenic resistant. 49,50 In other words, the metabolic resistance to cyflumetofen is mediated by a variety of detoxification enzymes in T. cinnabarinus. For instance, the phase I detoxification enzyme CCEs most likely contribute to cyflumetofen resistance by suppressing the activation of cyflumetofen to its active metabolite, AB-1.…”