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Molecular identification of mycorrhizal fungi in Dactylorhiza sambucina (Orchidaceae)

Biologia volume 64, pages 893–897 (2009)Cite this article

Abstract

We amplified and sequenced internal transcribed spacer (ITS) region of the nuclear ribosomal repeat from fungi in roots of Dactylorhiza sambucina (Orchidaceae) and used the extended database to identify the mycorrhizal fungi. We molecularly identified three ITS recovered from D. sambucina roots, one belonging to Rhizoctonia group, and two to ascomycetes, for the first time in Orchidoidae. In many cases, two sequence types were found from the same orchid root, providing that two taxa may be involved in mycorrhizal formation (multiple mycobiont colonization). Moreover, we demonstrated that D. sambucina plants, irrespective of their colour polymorphism, possess roots containing several fungi belonging to both asco- and basidiomycetes.

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References

  • Bidartondo M.I., Burghardt B., Gebauer G., Bruns T.D. & Read D.J. 2004. Changing partners in the dark: isotopic and molecular evidence of ectomycorrhizal liaisons between forest orchids and trees. Proc. R. Soc. London Series B-Biol. 271: 1799–1806.

    Article  CAS  Google Scholar 

  • Clements M.A. 1988. Orchid mycorrhizal associations. Lindleyana 3: 73–86.

    Google Scholar 

  • Currah R.S., Zettler L.W. & McInnis T.M. 1997. Epulorhiza inquilina sp. nov. from Platanthera (Orchidaceae) and a key to Epulorhiza species. Mycotaxon 61: 335–342.

    Google Scholar 

  • Curtis J.T. 1939. The relation of specificity of orchid mycorrhizal fungi to the problem of symbiosis. Am. J. Bot. 26: 390–399.

    Article  Google Scholar 

  • Doyle J.J. 1991. DNA protocols for plants — CTAB total DNA isolation, pp. 283–293. In: Hewitt G.M. & Johnston A. (eds), Molecular techniques in taxonomy. Springer Verlag, Berlin, Germany.

    Google Scholar 

  • Dressler R.L. 1981. The orchids: natural history and classification. Harvard University Press, Cambridge, MA, USA.

    Google Scholar 

  • Gardes M. & Bruns T.D. 1993. ITS primers with enhanced specificity for Basidiomycetes-application to the identification of mycorrhizae and rusts. Mol.Ecol. 2: 113–118.

    PubMed  Article  CAS  Google Scholar 

  • Girlanda M., Selosse M.A., Cafasso D., Brilli F., Delfine S., Fabbian R., Ghignone S., Pinelli P., Segreto R., Loreto F., Cozzolino S. & Perotto S. 2006 Inefficient photosynthesis in the Mediterranean orchid Limodorum abortivum is mirrored by specific association to ectomycorrhizal Russulaceae. Mol. Ecol. 15: 491–504.

    PubMed  Article  CAS  Google Scholar 

  • Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and program for Windows 95/98/NT. Nucl. Acids. Symp. Ser. 41: 95–98.

    CAS  Google Scholar 

  • Kjøller R. & Rosendahl S. 2000. Detection of arbuscular mycorrhizal fungi (Glomales) in roots by nested PCR and SSCP (single strand conformation polymorphism). Plant Soil 226: 189–196.

    Article  Google Scholar 

  • Kristiansen K.A., Taylor D.L., Kjøller R., Rasmussen H.N. & Rosendahl S. 2001. Identification of mycorrhizal fungi from single pelotons of Dactylorhiza majalis (Orchidaceae) using single-strand conformation polymorphism and mitochondrial ribosomal large subunit DNA sequences. Mol. Ecol. 10: 2089–2093.

    PubMed  Article  CAS  Google Scholar 

  • Kristiansen K.A., Freudenstein J.V., Rasmussen F.N. & Rasmussen H.N. 2004. Molecular identification of mycorrhizal fungi in Neuwiedia veratrifolia (Orchidaceae). Mol. Phylogenet. Evol. 33: 251–258.

    PubMed  Article  CAS  Google Scholar 

  • Leake J.R. 1994. The biology of myco-heterotrophic (’saprophytic’) plants. New Phytol. 127: 171–216.

    Article  Google Scholar 

  • Masuhara G. & Katsuya K. 1991. Fungal coil formation of Rhizoctonia repens in seedlings of Galeola septentrionalis (Orchidaceae). Bot. Mag. Tokyo 104: 275–281.

    Article  Google Scholar 

  • Masuhara G., Katsuya K. & Yamaguchi K. 1993. Potential for symbiosis of Rhizoctonia solani and binucleate Rhizoctonia with seeds of Spiranthes amoena var. amoena in vitro. Mycol. Res. 97: 746–752.

    Article  Google Scholar 

  • McCormick M.K., Whigham D.F. & O’Neill J. 2004. Mycorrhizal diversity in photosynthetic terrestrial orchids. New Phytol. 163: 425–438.

    Article  Google Scholar 

  • McKendrick S.L., Leake J.R., Taylor D.L. & Read D.J. 2002. Symbiotic germination and development of the myco-heterotrophic orchid Neottia nidus-avis in nature and its requirement for locally distributed Sebacina spp. New Phytol. 154:233–247.

    Article  Google Scholar 

  • Nilsson L.A. 1980. The pollination ecology of Dactylorhiza sambucina (Orchidaceae). Bot. Not. 133: 367–385.

    Google Scholar 

  • Otero J.T., Ackerman J.D. & Bayman P. 2002. Diversity and host specificity of endophytic Rhizoctonia -like fungi from tropical orchids. Am. J. Bot. 89: 1852–1858.

    Article  CAS  Google Scholar 

  • Perkins A.J. & McGee P.A. 1995. Distribution of the orchid mycorrhizal fungus Rhizoctonia solani, in relation to it’s host, Pterostylis acuminata, in the field. Aust. J. Bot. 43: 565–575.

    Article  Google Scholar 

  • Peterson R.L., Uetake Y. & Zelmer C. 1998. Fungal symbioses with orchid protocorms. Symbiosis 25: 29–55.

    Google Scholar 

  • Rasmussen H.N. 1995. Terrestrial orchids: from seed to mycotrophic plant. Cambridge University Press New York, NY, USA.

    Google Scholar 

  • Selosse M.A., Weiss M., Jany J.L. & Tillier A. 2002. Communities and populations of sebacinoid Basidiomycetes associated with the achlorophyllous orchid Neottia nidus-avis (L.) L.C.M. Rich. and neighbouring tree ectomycorrhizae. Mol. Ecol. 11: 1831–1844.

    PubMed  Article  CAS  Google Scholar 

  • Selosse M.A., Faccio A., Scappaticci G. & Bonfante P. 2004. Chlorophyllous and achlorophyllous specimens of Epipactis microphylla (Neottieae, Orchidaceae) are associated with ectomycorrhizal septomycetes, including truffles. Microb. Ecol. 47: 416–426.

    PubMed  Article  CAS  Google Scholar 

  • Shefferson R.P., Weiss M., Kull T. & Taylor D.L. 2005. High specificity generally characterizes mycorrhizal association in rare lady’s slipper orchids, genus Cypripedium. Mol. Ecol. 14: 613–626.

    PubMed  Article  CAS  Google Scholar 

  • Shefferson R.P., Kull T. & Tali K. 2008. Mycorrhizal interactions of orchids colonizing Estonian mine tailing hills. Am. J. Bot. 95: 156–164.

    Article  Google Scholar 

  • Smith S.E. & Read D.J. 1997. Mycorrhizal symbiosis. Academic Press, San Diego, CA, USA.

    Google Scholar 

  • Taylor D.L. & Bruns T.D. 1997. Independent, specialized invasions of ectomycorrhizal mutualism by two nonphotosynthetic orchids. Proc. Natl. Acad. Sci. USA 94: 4510–4515.

    PubMed  Article  CAS  Google Scholar 

  • Taylor D.L. & Bruns T.D. 1999. Community structure of ectomycorrhizal fungi in a Pinus muricata forest: minimal overlap between the mature forest and resistant propagule communities. Mol. Ecol. 8: 1837–1850.

    PubMed  Article  CAS  Google Scholar 

  • Taylor D.L. & McCormick M.K. 2008. Internal transcribed spacer primers and sequences for improved characterization of basidiomycetous orchid mycorrhizas. New Phytol. 177: 1020–1033.

    PubMed  Article  CAS  Google Scholar 

  • Taylor D.L., Bruns T.D., Leake J.R. & Read D.J. 2002. Mycorrhizal specificity and function in myco-heterotrophic plants, pp. 375–414. In: van der Heijden M.G.A.& Sanders R.I. (eds), The ecology of mycorrhizas. Springer Verlag, Berlin, Germany.

    Google Scholar 

  • Taylor D.L., Bruns T.D., Szaro T.M. & Hodges S.A. 2003. Divergence in mycorrhizal specialization within Hexalectris spicata (Orchidaceae), a nonphotosynthetic desert orchid. Am. J. Bot. 90: 1168–1179.

    Article  CAS  Google Scholar 

  • Tutin T.G., Heywood V.H., Burges N.A., Moore D.M., Valentine D.H., Walters S.M. & Webb D.A. 1980. Flora Europea. Cambridge Univ. Press, Cambridge, UK.

    Google Scholar 

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Authors and Affiliations

  1. Dipartimento di Ecologia, Universitè della Calabria, Arcavacata di Rende, I-87036, Cosenza, Italy

    Giuseppe Pellegrino & Francesca Bellusci

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  1. Giuseppe Pellegrino
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  2. Francesca Bellusci
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Correspondence to Giuseppe Pellegrino.

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Pellegrino, G., Bellusci, F. Molecular identification of mycorrhizal fungi in Dactylorhiza sambucina (Orchidaceae). Biologia 64, 893–897 (2009). https://doi.org/10.2478/s11756-009-0175-7

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  • DOI: https://doi.org/10.2478/s11756-009-0175-7

Key words

  • Dactylorhiza sambucina
  • ITS
  • mycorrhizal fungi
  • Orchidaceae