A Rapid and Cost-Effective Method for Genomic DNA Extraction from Plant Fungal Pathogens

Abstract

This chapter demonstrates a rapid, cost-effective, and safe method for extracting genomic DNA from multiple plant fungal pathogens. Pure cultures of Fusarium equesti, Neoscytalidium dimidiatum, Fusarium proliferatum, and Alternaria alternata, isolated from diseased wheat, grapevine, potato, and lily plants, respectively, were used. The fungal samples were ground with sterilised sand and 2N NaOH, followed by centrifugation to separate sand grains and fungal cellular components from the DNA. The resulting DNA was mixed with Tris buffer (1 M, pH 8). The internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) was successfully amplified, sequenced, and analysed from the extracted DNA of all four pathogens. This safe technique offers a straightforward, efficient, and non-hazardous method to obtain sufficient quantity and quality DNA suitable for molecular assays to identify plant fungi.