Multiple TaqMan qPCR and droplet digital PCR ( ddPCR ) diagnostics for pesticide resistance monitoring and management, in the major agricultural pest Tetranychus urticae

Autor: Chris Bass, John Vontas, Aris Ilias, Konstantinos Mavridis, Maria Riga, Kyriaki Maria Papapostolou, Thomas Van Leeuwen, Kleita Michaelidou, Anastasia Tsagkarakou
Rok vydání: 2021
Předmět:
Zdroj: Pest Management Science. 78:263-273
ISSN: 1526-4998
1526-498X
DOI: 10.1002/ps.6632
Popis: Background Decisions on which pesticide to use in agriculture are expected to become more difficult, as the number of available chemicals is decreasing. For Tetranychus urticae, a major pest for which a number of candidate markers for pesticide resistance are in place, molecular diagnostics could support decision-making for the rational use of acaricides. Results A suite of twelve of TaqMan qPCR assays [G314D (GluCl1), G326E and I321T (GluCl3), G119S, F331W (Ace-1), H92R (PSST), L1024V, F1538I (VGSC), I1017F (CHS1)], G126S, S141F, P262T (cytb)], were validated against Sanger-sequencing, and subsequently adapted for use with the ddPCR technology. The concordance correlation coefficient between the actual and ddPCR measured mutant allelic frequencies was 0.995 (95% CI = 0.991 - 0.998), and no systematic, proportional or random differences detected. The achieved Limit of Detection (LoD) was 0.1% (detection of a 1 mutant in a background of 999 wild type mites). The ddPCR assay panel was then assessed in terms of agreement with phenotypic resistance, through a pilot application in field populations from Crete, with strong correlation and thus predictive and diagnostic value of the molecular assays in some cases, e.g., etoxazole and abamectin resistance. Molecular diagnostics were able to capture incipient resistance that was missed by phenotypic bioassays in other cases. The molecular and phenotypic resistance screening of T. urticae field populations from Crete, revealed both multi-resistant and susceptible populations. Conclusion The highly sensitive T. urticae molecular diagnostic platforms developed in this study could prove a valuable tool for pesticide resistance management. This article is protected by copyright. All rights reserved.
Databáze: OpenAIRE
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