Development of a two‐step high‐resolution melting (HRM) analysis for screening sequence variants associated with resistance to the QoIs, benzimidazoles and dicarboximides in airborne inoculum of Botrytis cinerea. Issue 2 (22nd September 2014)

• Record Type: Journal Article
• Authors: Chatzidimopoulos, Michael; Ganopoulos, Ioannis; Vellios, Evangelos; Madesis, Panagiotis; Tsaftaris, Athanasios; Pappas, Athanassios C.
• Is Part Of: FEMS microbiology letters. Volume 360:Issue 2(2014:Nov.)
• Page Start: 126
• Page End: 131
• Publication Date: 2014-09-22
• Subjects: Microbiology -- Periodicals; 579
• DOI: 10.1111/1574-6968.12594
• Languages: English
• ISSNs: 0378-1097

Abstract

<abstract abstract-type="main" id="fml12594-abs-0001"> <title>Abstract</title> <p>A rapid, high‐resolution melting (HRM) analysis protocol was developed to detect sequence variations associated with resistance to the QoIs, benzimidazoles and dicarboximides in <italic>Botrytis cinerea</italic> airborne inoculum. HRM analysis was applied directly in fungal DNA collected from air samplers with selective medium. Three and five different genotypes were detected and classified according to their melting profiles in <italic>BenA</italic> and <italic>bos1</italic> genes associated with resistance to benzimidazoles and dicarboximides, respectively. The sensitivity of the methodology was evident in the case of the QoIs, where genotypes varying either by a single nucleotide polymorphism or an additional 1205‐bp intron were separated accurately with a single pair of primers. The developed two‐step protocol was completed in 82 min and showed reduced variation in the melting curves' formation. HRM analysis rapidly detected the major mutations found in greenhouse strains providing accurate data for successfully controlling grey mould.</p> </abstract>