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Fungal composition of lichen thalli assessed by single strand conformation polymorphism

Published online by Cambridge University Press:  03 June 2010

Lucia MUGGIA  and
Martin GRUBE
Lucia MUGGIA
Affiliation:
Institute of Plant Sciences, Karl-Franzens-University Graz, Holteigasse 6, 8010 Graz, Austria. Email: [email protected] Institute of Environmental Biotechnology, Graz University of Technology, Petersgasse 3, 8010 Graz, Austria.
Martin GRUBE
Affiliation:
Institute of Plant Sciences, Karl-Franzens-University Graz, Holteigasse 6, 8010 Graz, Austria. Email: [email protected]
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Abstract

Fungi that are unrelated to the mycobiont species frequently colonize lichens. Some of these fungal colonists are described lichenicolous fungi, lichen parasites and pathogens that produce recognizable morphological characters, while others apparently produce no noticeable structures. Here we apply the single strand conformation polymorphism (SSCP) technique to directly assess the abundance of different fungi in lichens. Twenty-eight lichen thalli were chosen, some with and some without externally visible symptoms of parasite infection, and these were subjected to total DNA extraction. PCR was conducted with fungal-specific primers for the ITS region of ribosomal DNA. Single strands of the products were separated on native acrylamide gels. The majority of lichen specimens, both infected and those without symptoms, displayed more than one band in the stained gels. In one case, 14 bands were detected using SSCP. Some of these bands apparently represent other neighbouring lichens in the habitat, but many are apparently non-lichen-forming. Since few lichen-associated fungi have been cultured and sequenced, it is difficult to know if SSCP bands represent obligate lichenicolous fungi, other asymptomatic lichen parasites, or fungi not obligately associated with lichens, but our results indicate that large numbers of non-lichen-forming fungi commonly co-occur with lichens in nature. For specimens of the filamentous lichens Cystocoleus ebeneus and Racodium rupestre we used cloned sequences to compare the number of sequences obtained by the SSCP method to the number obtained by direct sequencing of thallus extracts, and we generally found that more sequences could be detected by SSCP than could be seen by direct sequencing.

Keywords

diversityITSlichenicolous fungiSSCPsymbiosis
Type
Research Article
Information
The Lichenologist , Volume 42 , Issue 4 , July 2010 , pp. 461 - 473
Copyright
Copyright © British Lichen Society 2010

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References

Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research 25: 3389–3402.CrossRefGoogle ScholarPubMed
Anderson, I. C. & Cairney, J. W. G. (2004) Diversity and ecology of soil fungal communities: increased understanding through the application of molecular techniques. Environmental Microbiology 6: 769–779.CrossRefGoogle ScholarPubMed
Arnold, A. E., Miądlikowska, J., Higgins, K. L., Sarvate, S. D., Gugger, P., Way, A., Hofstetter, V., Kauff, K. & Lutzoni, F. (2009) A phylogenetic estimation of trophic transition networks for ascomycetous fungi: are lichens cradles of symbiotrophic fungal diversification? Systematic Biology 58: 283–297.CrossRefGoogle ScholarPubMed
Crittenden, P. D., David, J. C., Hawksworth, D. L. & Campbell, F. S. (1995) Attempted isolation and success in the culturing of a broad spectrum of lichen-forming and lichenicolous fungi. New Phytologist 130: 267–297.CrossRefGoogle Scholar
Cubero, O. F., Crespo, A., Fatehi, J. & Bridge, P. D. (1999) DNA extraction and PCR amplification method suitable for fresh, herbarium stored and lichenized fungi. Plant Systematics and Evolution 217: 243–249.CrossRefGoogle Scholar
Ekman, S. (1999) PCR optimization and troubleshooting, with special reference to the amplification of ribosomal DNA in lichenized fungi. Lichenologist 31: 517–531.CrossRefGoogle Scholar
Gardes, M. & Bruns, T. D. (1993) ITS primers with enhanced specificity for basidiomycetes. Application for the identification of mycorrhizae and rust. Molecular Ecology 2: 113–118.CrossRefGoogle Scholar
Girlanda, M., Isocrono, D., Bainco, C. & Luppi-Mosca, A. M. (1997) Two foliose lichens as microfungal ecological niches. Mycologia 89: 531–536.CrossRefGoogle Scholar
Hall, T. A. (1999) BioEdit: a user friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acid Symposium Series 41: 95–98.Google Scholar
Harutyunyan, S., Muggia, L. & Grube, M. (2008) Black fungi in lichens from seasonally arid habitats. Studies in Mycology 61: 83–90.CrossRefGoogle ScholarPubMed
Hawksworth, D. L. & Honegger, R. (1994) The lichen thallus: a symbiotic phenotype of nutritionally specialized fungi and its response to gall producer. In Plant Galls: Organisms, Interactions, Populations. (Williams, M. A. I., ed.): 77–98. Oxford: Clarendon Press.CrossRefGoogle Scholar
Huelsenbeck, J. P. & Ronquist, F. (2003) MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics 19: 1572–1574.Google Scholar
Lawrey, J. D. (1993) Chemical ecology of Hobsonia christiansenii, a lichenicolous hyphomycete. American Journal of Botany 80: 1109–1113.CrossRefGoogle Scholar
Lawrey, J. D. & Diederich, P. (2003) Lichenicolous fungi: interactions, evolution, and biodiversity. Bryologist 106: 80–120.CrossRefGoogle Scholar
Mitchell, J. I. & Zuccaro, A. (2006) Sequences, the environment and fungi. Mycologist 20: 62–74.CrossRefGoogle Scholar
Muggia, L., Hafellner, J., Wirtz, N., Hawksworth, D. L. & Grube, M. (2008) The sterile microfilamentous lichenized fungi Cystocoleus ebeneus and Racodium rupestre are relatives of plant pathogens and clinically important dothidealean fungi. Mycological Reserach 112: 50–56.CrossRefGoogle ScholarPubMed
Orita, M., Iwahan, H., Kanazawa, H., Hayashi, K. & Sekiya, T. (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proceedings of the National Academy of Sciences USA 86: 2766–2770.CrossRefGoogle ScholarPubMed
Page, R. D. M. (1996) TREEVIEW: an application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences 12: 357–358.Google ScholarPubMed
Petrini, O., Hakem, U. & Dreyfuss, M. M. (1990) An analysis of fungal communities isolated from fruticose lichens. Mycologia 82: 444–451.CrossRefGoogle Scholar
Posada, D. & Crandall, K. A. (1998) Modeltest – testing the model of DNA substitution. Bioinformatics 14: 817–818.CrossRefGoogle ScholarPubMed
Prillinger, H., Kraepelin, G., Lopandic, K., Schweigkofler, W., Molnar, O., Weigang, F. & Dreyfuss, M. M. (1997) New species of Fellomyces isolated from epiphytic lichen species. Systematic and Applied Microbiology 20: 572–574.CrossRefGoogle Scholar
Redman, R. S., Dunigan, D. D. & Rodriguez, R. J. (2001) Fungal symbiosis: from mutualism to parasitism, who controls the outcome, host or invader? New Phytologist, 151: 705–716.CrossRefGoogle ScholarPubMed
Rodriguez, F., Oliver, J. L., Marin, A. & Medina, J. R. (1990) The general stochastic model of nucleotide substitution. Journal of Theoretical Biology 142: 485–501.CrossRefGoogle ScholarPubMed
Ronquist, F., Huelsenbeck, J. P. & van der Mark, P. (2005) MrBayes 3.1 Manual, http://mrbayes.csit.fsu.edu/mb3.1_manual.pdf.Google Scholar
Ruibal, C., Platas, G. & Billis, G. F. (2005) Isolation and characterization of melanized fungi from limestone formations in Mallorca. Mycological Progress 4: 23–38.CrossRefGoogle Scholar
Seifert, K. A. (2009) Progress towards DNA barcoding of fungi. Molecular Ecology Resources 9 (suppl. 1): 83–89.CrossRefGoogle ScholarPubMed
Sikaroodi, M., Lawrey, J. D., Hawksworth, D. L. & DePriest, P. (2001) The phylogenetic position of selected lichenicolous fungi: Hobsonia, Illosporium, and Marchandiomyces. Mycological Research 105: 453–460.CrossRefGoogle Scholar
Torzilli, A. P., Balakrishna, S., O'Donnell, K. & Lawrey, J. D. (2002) The degradative activity of a lichenicolous Fusarium sp. compared to related entomogenous species. Mycological Research 106: 1204–1210.CrossRefGoogle Scholar
White, T. J., Bruns, T. D., Lee, S. & Taylor, J. (1990) Amplification and direct sequencing of fungal ribosomal DNA genes for phylogenies. In PCR Protocols, a Guide to Methods and Applications. (Innis, M. A., Gelfand, D. H., Snisky, J. J., & White, T. J., eds): 315–322. San Diego: Academic Press.Google Scholar
Wolinski, H., Grube, M. & Blanz, P. (1999) Direct PCR of symbiotic fungi using microslides. Biotechniques 26: 454–455.CrossRefGoogle ScholarPubMed