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A rapid and efficient protocol for the extraction of genomic DNA from plant pathogenic fungi was developed. Key features of the protocol include the SDS-assisted lysis of fungal mycelium with inclusion of a glass bead to help break hyphal walls, followed by isopropanol precipitation of the DNA. The protocol was used to extract genomic DNA from a collection of 26 fungal species, representing many important plant pathogens. Yield of DNA ranged from 2.1-4.9 μg per 20 mg of mycelium or 0.4-0.6 μg per 20 mg of spores. The DNA was of sufficient purity to be digested by restriction enzymes, to serve as a template in the PCR-amplification of genomic fragments as large as 4.9 kb, and to be used in dot-blot hybridization for the detection of multiple- and single-copy genes. [ABSTRACT FROM AUTHOR]

Examines the internal fruit rot caused by Fusarium subglutinans in sweet peppers. Development of a dot-blot hybridization technique for rapid and accurate detection and diagnosis of the plant disease; Differentiation of the disease from fruit rots of peppers caused by other pathogens; Capability of the dot-blot assay to detect F. subglutinans infection in the frozen and fresh host tissues.