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Published on: June 25, 2020
Detection of Botrytis cinerea by loop-mediated isothermal amplification
J A Tomlinson1, M J Dickinson, N Boonham
1The Food and Environment Research Agency, Sand Hutton, York, UK. jenny.tomlinson@fera.gsi.gov.uk
Letters in Applied Microbiology
|October 30, 2010
Summary
A new loop-mediated isothermal amplification (LAMP) method rapidly and accurately detects Botrytis cinerea, a fungal pathogen. This sensitive assay is suitable for on-site detection in plant material, aiding in crop management.
Area of Science:
- Plant pathology
- Molecular diagnostics
- Mycology
Background:
- Botrytis cinerea causes significant post-harvest losses in various commodities.
- Accurate and rapid detection methods are crucial for effective management of B. cinerea.
Purpose of the Study:
- To develop a sensitive, rapid, and simple loop-mediated isothermal amplification (LAMP) assay for Botrytis cinerea detection.
- To create a method suitable for use outside conventional laboratory settings.
Main Methods:
- A LAMP assay was designed targeting the intergenic spacer of the B. cinerea nuclear ribosomal DNA (rDNA).
- The assay's sensitivity and specificity were characterized using extracted DNA and tested against other fungal pathogens.
- Detection was performed using a real-time LAMP platform (OptiGene Genie I).
Main Results:
- The LAMP assay demonstrated high sensitivity, consistently amplifying 65 pg of B. cinerea DNA.
- No cross-reactivity was observed with other fungal pathogens, except for the closely related Botrytis pelargonii.
- B. cinerea was detected in infected rose petals within 15 minutes using the real-time LAMP platform.
Conclusions:
- The developed LAMP assay is effective for the rapid detection of B. cinerea in infected plant material.
- The method's sensitivity, specificity, simplified equipment, and reduced reaction time make it suitable for on-site applications.
- This assay can inform decisions on storage and processing of commodities affected by B. cinerea.
