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A new endophytic insect-associated Daldinia species, recognised from a comparison of secondary metabolite profiles and molecular phylogeny

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Abstract

Xylariaceous endophytes are commonly encountered in plants, and it has recently been found out that insect vectors play a major role in their life cycle, as well as in the apparent host-specificity of the fungal stromata. In this study, a new insect-associated, endophytic anamorph species of Daldinia is recognised from a comparison of a) morphological and ecological data; b) molecular phylogenetic data based on rDNA, alpha-actin and beta-tubulin genes; and c) secondary metabolite profiles based on high performance liquid chromatography/diode array/mass spectrometry (HPLC-DAD/MS) and gas chromatography/mass spectrometry (GC-MS) in comparison with authentic strains of Daldinia and other Xylariaceae. The new species, Daldinia hawksworthii, is a symbiont of the willow woodwasp, Xiphydria prolongata, which is in turn preferentially associated with Salix. A comparison of HPLC-DAD/MS profiles from standardised cultures revealed the presence of a specific compound, which was not encountered in any of the ca. 250 Daldinia strains studied for comparison. This compound was isolated after fermentation of the fungus in 10 l scale by preparative chromatography. Structure elucidation by NMR spectroscopy revealed a novel pyrone, for which the name dalsymbiopyrone is proposed. The compound showed weak antimicrobial and cytotoxic effects and might be a signal molecule involved in the trilateral ecological interaction between the host plant, the insect, and its fungal symbiont.

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Acknowledgments

We are grateful to Beata Schmieschek and Bärbel Köpcke (formerly InterMed Discovery) for performing initial experiments on HPLC profiling and fermentation of selected D. hawksworthii strains. Derek Persoh (Bayreuth) is thanked for providing a preliminary phylogeny by comparing the manifold unpublished data from previous work on Xylariaceae with the sequence data of the new species, and Alberto Plaza (HIPS, Saarbrücken) is thanked for his invaluable help in recording NMR spectra. We are extremely grateful to Aileen Teichmann (HZI) for performing exhaustive HPLC-DAD/MS analyses, and to Wolfgang Kessler (HZI) and his team for performing the 10 l fermentation that finally allowed for isolation of dalsymbiopyrone. Ajda Médjedovic and Hans-Josef Schroers are thanked for providing the Colletotrichum test strain.

Last but not least we gratefully acknowledge support from COST Action FA1103 “Endophytes in Biotechnologiy and Agriculture”.

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Authors and Affiliations

  1. Institute of Microbiology of the ASCR, v.v.i., Vídeňská 1083, 14220, Prague 4, Czech Republic

    Sylvie Pažoutová

  2. Department Microbial Drugs, Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstrasse 7, 38124, Braunschweig, Germany

    Stefanie Follert, Matthias Keck, Frank Surup & Marc Stadler

  3. IMD Natural Solutions GmbH, Otto-Hahn-Strasse 15, 44227, Dortmund, Germany

    Jens Bitzer

  4. Faculty of Forestry and Wood Sciences, Czech University of Life Sciences, Kamýcká 129, 165 21, Prague 6, Czech Republic

    Petr Šrůtka & Jaroslav Holuša

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  1. Sylvie Pažoutová
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  2. Stefanie Follert
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Correspondence to Marc Stadler.

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Pažoutová, S., Follert, S., Bitzer, J. et al. A new endophytic insect-associated Daldinia species, recognised from a comparison of secondary metabolite profiles and molecular phylogeny. Fungal Diversity 60, 107–123 (2013). https://doi.org/10.1007/s13225-013-0238-5

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