Pest Management Science (2009) 65, 413-419

Changjun Chen, Wei Zhao, Yuejian Lu, Jianxin Wang, Yu Chen, Hongxia Li and Mingguo Zhou (2009)
High-throughput detection of highly benzimidazole-resistant allele E198A with mismatch primers in allele-specific real-time polymerase chain reaction
Pest Management Science 65 (4), 413-419
Abstract: BACKGROUND: A point mutation often confers resistance of organisms against medical drugs and agricultural pesticides. Allele-specific nucleotide polymerase chain reaction (ASPCR) and allele-specific quantitative real-time PCR using SYBR Green (ASQPCR) are widely and effectively applied to detect and monitor this type of resistance. However, the former is unsuitable for high-throughput detection, and the latter often reduces the accuracy of detection.
RESULTS: In order to decrease background amplification, a rapid and high-throughput genotyping method with mismatch primers was developed (ASQPCR-MP) and applied specifically to survey the frequency of the highly benzimidazole-resistant MBC^HR mutation (E198A) in the ß-tubulin gene of Sclerotinia sclerotiorum (Lib.) de Bary populations. Genomic DNA from 223 sclerotia was analysed. Similar genotype results were also obtained using ASPCR with mismatch primers and a mycelial growth inhibition assay. It was found that ASQPCR-MP clearly differentiated MBC^HR and benzimidazole-sensitive MBC^S phenotypes. Moreover, ASQPCR-MP took less than 6 h to complete.
CONCLUSION: ASQPCR-MP appears suitable for large epidemiological studies involving resistant genotypes and requiring high-throughout formats.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Ming-Guo Zhou, Chang-Jun Chen

Research topic(s) for pests/diseases/weeds:
pesticide resistance of pest

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Sclerotinia sclerotiorum