Genotyping of benzimidazole-resistant and dicarboximide-resistant mutations in Botrytis cinerea using real-time polymerase chain reaction assays

被引:91
|
作者
Banno, Shinpei [2 ]
Fukumori, Fumiyasu [1 ]
Ichiishi, Akihiko [1 ]
Okada, Kiyotsugu [3 ]
Uekusa, Hidetoshi [4 ]
Kimura, Makoto [5 ]
Fujimura, Makoto [1 ]
机构
[1] Toyo Univ, Fac Life Sci, Gunma 3740193, Japan
[2] Toyo Univ, Plant Funct Res Ctr, Gunma 3740193, Japan
[3] Agr Food & Environm Sci Res Ctr Osaka Prefecture, Osaka 5830862, Japan
[4] Kanagawa Prefectural Agr Res Inst, Hiratsuka, Kanagawa 2591204, Japan
[5] RIKEN, Discovery Res Inst, Plant & Microbial Metab Engn Res Unit, Wako, Saitama 3510198, Japan
关键词
Botryotinia fuckeliana;
D O I
10.1094/PHYTO-98-4-0397
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Botrytis cinerea, an economically important gray mold pathogen, frequently exhibits multiple fungicide resistance. A fluorescence resonance energy transfer-based real-time polymerase chain reaction assay has been developed to detect berizimidazole- and dicarboximide-resistant mutations. Three benzimidazole-resistant mutations-(198)Glu to Ala (E198A), F200Y, and E198K-in beta-tubulin BenA were detected using a single set of fluorescence-labeled sensor and anchor probes by melting curve analysis. Similarly, three dicarboximide-resistant mutations-1365S, V368F plus Q369H, and Q369P-in the histidine kinase BcOS1 were successfully distinguished. Unassigned melting profiles in BenA genotyping assay resulted in the identification of a new benzimidazole-resistant BenA E198V mutation. This mutation conferred resistance to carbendazim as do E198A and E198K mutations. The isolates with BenA E198V mutation showed a negative cross-resistance to diethofencarb, but to a lesser extent than the E198A mutants. A survey of 210 B. cinerea field isolates revealed that most of benzimidazole-resistant isolates possessed the E198V or E198A mutation in the BenA gene, and the 1365S mutation in the BcOS1 gene was also frequently observed in Japanese isolates. However, benzimidazole-resistant isolates with BenA F200Y or E198K mutations, which confer the diethofencarb-insensitive phenotype, were rare., Our BenA and BcOS1 genotyping is a rapid and reliable method that is suitable for monitoring the fungicide-resistant field population.
引用
收藏
页码:397 / 404
页数:8
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