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

Changes in gene expression are associated with the evolution of pesticide resis-tance 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 1685 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 in-terference 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,in-cluding 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(LC2o)of cyflumetofen.However,16 detoxification enzyme genes were up-regulated by 1-time selection with LCso of cyflumetofen.Interestingly,the 16 genes were overexpressed in all 3 resistance stages.These results indicated that 1685 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 de-velopment 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.