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Chlorophyllous and Achlorophyllous Specimens of Epipactis microphylla (Neottieae, Orchidaceae) Are Associated with Ectomycorrhizal Septomycetes, including Truffles

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Abstract

Mycoheterotrophic species (i.e., achlorophyllous plants obtaining carbon from their mycorrhizal fungi) arose many times in evolution of the Neottieae, an orchid tribe growing in forests. Moreover, chlorophyllous Neottieae species show naturally occurring achlorophyllous individuals. We investigated the fungal associates of such a member of the Neottieae, Epipactis microphylla, to understand whether their mycorrhizal fungi predispose the Neottieae to mycoheterotrophy. Root symbionts were identified by sequencing the fungal ITS of 18 individuals from three orchid populations, including achlorophyllous and young, subterranean individuals. No rhizoctonias (the usual orchid symbionts) were recovered, but 78% of investigated root pieces were colonized by Tuber spp. Other Pezizales and some Basidiomycetes were also found. Using electron microscopy, we demonstrated for the first time that ascomycetes, especially truffles, form typical orchid mycorrhizae. All identified fungi (but one) belonged to taxa forming ectomycorrhizae on tree roots, and four of them were even shown to colonize surrounding trees. This is reminiscent of mycoheterotrophic orchid species that also associate with ectomycorrhizal fungi, although with higher specificity. Subterranean and achlorophyllous E. microphylla individuals thus likely rely on tree photosynthates, and a partial mycoheterotrophy in individuals plants can be predicted. We hypothesize that replacement of rhizoctonias by ectomycorrhizal symbionts in Neottieae entails a predisposition to achlorophylly.

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Acknowledgments

We are grateful to F. Dusak and P. Pernot (Société Française d’Orchidophilie) for access to population C. We thank A. Dettai, A. Tillier and S. Tillier (Muséum National d’Histoire Naturelle), as well as D. Marsh and C. Saison for their constant help. We thank M. Bidartondo for helpful suggestions and providing the ITS4tul primer. We also thank an anonymous referee for careful remarks on the submitted version of this paper. The research was funded by the Muséum National d’Histoire Naturelle (Service de Systématique Moléculaire) and the Société Française d’Orchidophilie (M.-A. Selosse) and the Strategic Programme CNR-Regioni on Tuber (P. Bonfante).

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Authors and Affiliations

  1. UMR CNRS 7138 “Systématique, Adaptation et Evolution’’ and Service de Systématique Moléculaire (IFR CNRS 101), Muséum National d’Histoire Naturelle, 43, rue Cuvier, 75005, Paris, France

    M.-A. Selosse

  2. Dipartimento di Biologia Vegetale dell’Università, Istituto per la Protezione delle Piante—CNR, Viale Mattioli, 25, 10125, Torino, Italy

    A. Faccio & P. Bonfante

  3. Société Française d’Orchidophilie, Les Combeaux, Quartier Les Rouvières, 26220, Dieulefit, France

    G. Scappaticci

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  1. M.-A. Selosse
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  4. P. Bonfante
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Correspondence to M.-A. Selosse.

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Selosse, MA., Faccio, A., Scappaticci, G. et al. Chlorophyllous and Achlorophyllous Specimens of Epipactis microphylla (Neottieae, Orchidaceae) Are Associated with Ectomycorrhizal Septomycetes, including Truffles. Microb Ecol 47, 416–426 (2004). https://doi.org/10.1007/s00248-003-2034-3

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