Alkaline Extraction of DNA from Pathogenic Fungi for PCR-RFLP Analysis

For the preparation of DNA samples from fungal mycelia alkaline extraction method was applied and assessed its usefulness for PCR-RFLP analysis. Using alkaline treatment protocols, 18S ribosomal DNAs (rDNA) derived from fungal genomic DNA of Pyricularia oryzae, P. zingiberi, Rhizoctonia solani and R. oryzae were PCR-amplified and digested with Hha I, Msp I and Hae III. RFLP analysis with HhaI showed the divergent polymorphism between genus Pyricularia and Rhizoctonia. The alkaline DNA extraction method saves the time and labor required for DNA sample preparation and seemed to be useful for rapid identification of specific fungal species by PCR-RFLP analysis.