Fast Microwave-based DNA Extraction from Vegetative Mycelium and Fruiting Body Tissues of Agaricomycetes for PCR Amplification

Order of Publishing in Issue: 
1
Volume :7
Issue :4
Regular
October, 2013
Page No: 
825-836
Authors: 
Bastian Dörnte, Ursula Kües
Address: 
Büsgen-Institute, Division of Molecular Wood Biotechnology and Technical Mycology, University of Goettingen, Büsgenweg 2, 37077 Goettingen, Germany *For correspondence – ukuees@gwdg.de
Email-ID: 
ukuees@gwdg.de

In this study, we tested a microwave-based DNA extraction method for subsequent DNA amplifications by PCR on vegetative mycelia and mushrooms of different Agaricomycetes. The extraction method requires tiny amounts of fungal material, is rapid and achieved within minutes, why it is superior to classical extraction methods which are work-intensive and require larger amounts of starting material, hours of time for performance and in addition specific expensive and hazardous chemicals for cell lysis and DNA purification. The microwave method with highest reliability is suitable for vegetative mycelium harvested from fresh and also from aged fungal cultures. It is especially attractive for slowgrowing species of which larger amounts of mycelium are difficult to obtain from. The method is further applicable with success rates between 76.9% and 90.9% to fleshy mushrooms over a wide range of families of Agaricomycetes, both in fresh as well as in dried form, and also to firm young and older fruiting bodies of more robust leathery, corky and woody textures. Also noncultivable species can thus be accessed for DNA analyses. Finally, we show that also fungal infested plant material such as millet, straw, wood and bark can be used. However, here the success depends on freshness of the material and on presence of sufficient surface mycelium.

Keywords: 
Agaricomycetes, mushrooms, mycelium, microwaving, DNA extraction, PCR amplification
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