Mycological Progress

, Volume 13, Issue 3, pp 849–856 | Cite as

New isolation method for endophytes based on enzyme digestion

  • René Prior
  • Katharina Görges
  • Andrey Yurkov
  • Dominik Begerow
Original Article


Investigation of the diversity and ecology of endophytic fungi has gained popularity in recent decades. Thereby, culture-independent methods were very instrumental as they revealed greater species richness and challenged the efficiency of isolation methods. However, culture-based methods remain the only possibility to provide strains for future studies. To improve the efficiency of endophyte isolation, we compared seven different artificial media that included a variety of plant extracts. Furthermore, we developed a method based on enzymatic digestion of plant tissue to improve the isolation efficiency in terms of species richness. The effect of additional plant extract in media that contained yeast and malt extract was of minor significance, whereas isolations using enzymatic digestion of plant leaf tissue combined with a 1:10 dilution of nutrients revealed a much higher diversity of species. Using this protocol, we isolated three times as many species from the same leaves than with previously described methods. This is reflected by the large number of singletons obtained in culture-independent approaches to study the diversity of endophytic fungi.


Endophytes Fungi Strain cultivation Isolation method Enzyme digestion 



We thank Danja Schünemann for providing principles and recipes of the enzymatic digestion method. We thank Dominik Schmidt, Moritz Mittelbach, Alistair McTaggart and Derek Peršoh for critically reading the manuscript and for their advice. We thank Ilse Weßel and Tanja Rollnik for technical support and the Deutsche Bundesstiftung Umwelt for financial support.


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Copyright information

© German Mycological Society and Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • René Prior
    • 1
  • Katharina Görges
    • 1
  • Andrey Yurkov
    • 2
  • Dominik Begerow
    • 1
  1. 1.Geobotany, Department of Evolution and Biodiversity of Plants, Faculty of Biology and BiotechnologyRuhr-Universität BochumBochumGermany
  2. 2.Leibniz Institute DSMZ-German Collection of Microorganisms and Cell CulturesBrunswickGermany

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