VGSC mutations and P450 overexpression are associated with beta-cypermethrin resistance in Aphis gossypii Glover from a Chinese wolfberry (Lycium barbarum L.) field
Identification and characterization of genes and target-site mutations associated with beta-cypermethrin resistance in Aphis gossypii Glover collected from a Chinese wolfberry (Lycium barbarum L.) field. we collected a beta-cypermethrin resistant A. gossypii strain (HSP) from a Chinese wolf-berry orchard in a major growing area of Ningxia wolfberry (Wuzhong city). Subsequently, to elucidate the potential roles of P450s, CarEs and GSTs in beta-cypermethrin resistance in the A. gossypii strain, we performed synergistic bioassays, as well as enzyme activity assays, to confirm their effects. Further, we carried out a comparative transcriptome anal-ysis to identified the overexpression of detoxification enzyme genes associated with the beta-cypermethrin resistance. According to the transcriptome variations, we also meas-ured the expression levels of the upregulated P450s genes involved in beta-cypermethrin resistance in the A. gossypii resistant strain, using a quantitative real-time PCR assay. Moreover, the potential mutations in VGSC genes and their frequencies were detected to reveal the VGSC genotype of the resistant strain. Overall design: A highly beta-cypermethrin-resistant Aphis gossypii population was found in the field while high-throughput RNA-seq was used to determine the transcriptome profile of beta-cypermethrin-resistant (RGL) and beta-cypermethrin-sensitive (SGL) A. gossypii without exposure to beta-cypermethrin. Library construction and sequencing were conducted with three replicates for each pooled strain individuals. The cDNA library sequencing was performed by an Illumina HiSeq 2500 sequencing platform generating paired-end reads.