Rapid PCR-based method for the direct analysis of fungal communities in complex environmental samples

Fungal community analysis using 18S rDNA primer pairs and denaturing gradient gel electrophoresis of PCR products (Vainio, E.J., Hantula, J., 2000. Mycological Research 104, 927–936) was applied to field studies of the forest ecosystem. We report a DNA extraction method producing high quality DNA allowing successful PCR amplification from problematic samples without use of nested polymerase chain reaction (PCR) procedures. The analysis was found to be applicable for samples from environments of varying fungal diversities and high organic matter content: wood samples from fallen branches of trees, laboratory mini-ecosystems and forest humus samples. When the method was tested using replicate forest soil samples, it was shown to be highly reproducible.