Affiliation:
1. Institute of Plant Protection Beijing Academy of Agriculture and Forestry Sciences Beijing China
2. Department of Entomology and MOA Key Lab of Pest Monitoring and Green Management, College of Plant Protection China Agricultural University Beijing China
3. Beijing Engineering and Technique Research Center for Hybrid Wheat Beijing Academy of Agriculture and Forestry Sciences Beijing China
4. School of BioSciences Bio21 Institute, University of Melbourne Parkville Australia
Abstract
AbstractBACKGROUNDPesticide resistance is a long‐standing and growing problem in the chemical control of invertebrate pests. Molecular diagnostic methods can facilitate pesticide resistance management by accurately and efficiently detecting resistant mutations and their frequency. In this study, the kompetitive allele specific PCR (KASP) approach, a technology for high‐throughput single nucleotide polymorphism (SNP) genotyping, is validated as a useful method for characterizing genotypes at a pesticide‐resistance locus for the first time. We focus on the spinetoram resistance mutation of G275E in the nicotinic acetylcholine receptor alpha 6 (nAChR α6) subunit gene of Thrips palmi.RESULTSOf the 341 individuals of Thrips palmi tested, 98.24% were successfully genotyped, with 100% concordance with Sanger sequencing results. We then quantitatively mixed genomic DNA of known genotypes to establish 21 DNA mixtures with a resistant allele frequency ranging from 0 to 100% at steps of 5%. The linear discriminant analysis (LDA) showed that 75.8% of original grouped cases were correctly classified; six groups had no overlap in membership (resistant allele frequency: 0%, 5%, 10–75%, 80–85%, 90–95%, and 100%). When we chose 11 pooled samples with 10% steps for LDA, 84.4% of original grouped cases were correctly classified; seven groups had no overlap in membership (0%, 10%, 20–30%, 40–70%, 80%, 90%, 100%). The results indicated that KASP applied to pooled samples may provide a semi‐quantitative estimate of resistance.CONCLUSIONSOur study points to the suitability of KASP for high‐throughput genotyping of genotypes affecting pesticide resistance and semi‐quantitative assessments of resistance allele frequencies in populations. © 2023 Society of Chemical Industry.
Funder
Beijing Natural Science Foundation
Key Research and Development Program of Jiangxi Province
Subject
Insect Science,Agronomy and Crop Science,General Medicine
Cited by
2 articles.
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