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BACKGROUND: Insecticide resistance has emerged in various western flower thrips (WFT) populations across the world, threatening the efficiency of chemical control applications. Elucidation of insecticide resistance mechanisms at the molecular level provides markers for the development of diagnostics, to monitor the trait and support evidence-based resistance management. RESULTS: TaqMan and Droplet Digital PCR (ddPCR) diagnostics were developed and validated, against Sanger sequencing, in individual and pooled WFT samples, respectively, for the: G275E mutation (nicotinic acetylcholine receptor α6 gene- nAChR α6) associated with resistance to nAChR allosteric modulators - site I (spinosyns), L1014F, T929I, T929C and T292V mutations (voltage-gated sodium channel gene- vgsc) linked with pyrethroid resistance and I1017M (chitin synthase 1 gene- chs1) conferring resistance to growth inhibitors affecting CHS1 (benzoylureas). The detection limits of ddPCR assays for mutant allelic frequencies (MAFs) were in the range of 0.1%-0.2%. The assays were applied in nine WFT field populations from Crete, Greece. The G275E (MAF = 29.66%-100.0%), T929I and T929V (combined MAFs = 100%), L1014F (MAF = 11.01%-37.29%) and I1017M (MAF = 17.74%-51.07%) mutations, were present in all populations. CONCLUSION: The molecular diagnostics panel that was developed in this study can facilitate the quick and sensitive pesticide resistance monitoring of WFT populations at the molecular level, to support evidence-based IRM strategies. |
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