Application of DNA-based genotyping techniques for the detection of kdr-like pyrethroid resistance in field populations of Colorado potato beetle

Permethrin resistance in 16 field populations of Colorado potato beetle was established by bioassay. Permethrin-resistant genotypes were detected using two genotyping techniques, bidirectional polymerase chain reaction amplification of specific alleles (bi-PASA) and single stranded conformational polymorphism (SSCP). The bi-PASA technique was 86% and SSCP was 95% reliable. In terms of accuracy, bi-PASA determined the correct genotype compared to the actual sequencing data 98% and SSCP 93% of the time. There was marked correlation between increasing survivorship (increasing LD50values) and increasing proportions of resistant genotypes (increasing percentage of resistant allele). These techniques allowed the detection of a resistant allele 98% of the time when sampling only 384 beetles from an overwintering adult population (4995 beetles/ha) that possessed a 1% resistant allele frequency at a cost of $500/ha, substantiating the cost effectiveness of the genotyping approaches to monitoring.