Amplicon sequencing analysis using Illumina MiSeq for monitoring a cyantraniliprole resistance-related mutation in the diamondback moth populations

To monitor the development of cyantraniliprole resistance in the diamondback moth, Plutella xylostella (L.) (Lepidoptera; Plutellidae), we conducted an amplicon sequencing analysis using Illumina MiSeq for diamondback moth individuals collected in Nagano prefecture, Japan in 2016 and 2019. A mutation site related to cyantraniliprole resistance was amplified from genomic DNA of the collected diamondback moth individuals and genotyped by the amplicon sequencing (mutation sites related to resistance of other major insecticides including diamides, pyrethroids, Bt toxins, organophosphates, spinosyns and indoxacarb were also amplified in the experiment). The genotyping result was used as real data for evaluating sensitivity and specificity of the multiplex PCR-based molecular diagnostic method which was developed by us for simply and qucikly genotyping the mutation site of cyantraniliprole resistance in the diamondback moth.