Simultaneous Detection of Multiple Benzimidazole-Resistant β-Tubulin Variants of Botrytis cinerea using Loop-Mediated Isothermal Amplification

Optimal disease management depends on the ability to monitor the development of fungicide resistance in plant pathogen populations. Benzimidazole resistance is caused by the point mutations of the β-tubulin gene in Botrytis cinerea, and three mutations (E198A, E198K, and E198V) at codon 198 account...

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Veröffentlicht in:Plant disease. - 1997. - 102(2018), 10 vom: 21. Okt., Seite 2016-2024
1. Verfasser: Duan, Ya Bing (VerfasserIn)
Weitere Verfasser: Yang, Ying, Wang, Jian Xin, Chen, Chang Jun, Steinberg, Gero, Fraaije, Bart A, Zhou, Ming Guo
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2018
Zugriff auf das übergeordnete Werk:Plant disease
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Anthelmintics Benzimidazoles Tubulin benzimidazole E24GX49LD8
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245 1 0 |a Simultaneous Detection of Multiple Benzimidazole-Resistant β-Tubulin Variants of Botrytis cinerea using Loop-Mediated Isothermal Amplification 
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520 |a Optimal disease management depends on the ability to monitor the development of fungicide resistance in plant pathogen populations. Benzimidazole resistance is caused by the point mutations of the β-tubulin gene in Botrytis cinerea, and three mutations (E198A, E198K, and E198V) at codon 198 account for more than 98% of all resistant strains. Although traditional methods remain a cornerstone in monitoring fungicide resistance, molecular methods that do not require the isolation of pathogens can detect resistance alleles present at low frequencies, and require less time and labor than traditional methods. In this study, we present an efficient, rapid, and highly specific method for detecting highly benzimidazole-resistant B. cinerea isolates based on loop-mediated isothermal amplification (LAMP). By using specific primers, we could simultaneously detect all three resistance-conferring mutations at codon 198. The LAMP reaction components and conditions were optimized, and the best reaction temperatures and times were 60 to 62°C and 45 min, respectively. When B. cinerea field isolates were assessed for benzimidazole resistance, similar results were obtained with LAMP, minimal inhibition concentration, and sequencing. The LAMP assay developed in the current study was highly suitable for detection of highly benzimidazole-resistant field isolates of B. cinerea 
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650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a Benzimidazoles  |2 NLM 
650 7 |a Tubulin  |2 NLM 
650 7 |a benzimidazole  |2 NLM 
650 7 |a E24GX49LD8  |2 NLM 
700 1 |a Yang, Ying  |e verfasserin  |4 aut 
700 1 |a Wang, Jian Xin  |e verfasserin  |4 aut 
700 1 |a Chen, Chang Jun  |e verfasserin  |4 aut 
700 1 |a Steinberg, Gero  |e verfasserin  |4 aut 
700 1 |a Fraaije, Bart A  |e verfasserin  |4 aut 
700 1 |a Zhou, Ming Guo  |e verfasserin  |4 aut 
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