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Structural characterization and molecular identification of arbuscular mycorrhiza morphotypes of Alzatea verticillata (Alzateaceae), a prominent tree in the tropical mountain rain forest of South Ecuador

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Abstract

The vast majority of the highly diverse trees in the tropical mountain rain forest of South Ecuador form arbuscular mycorrhizas, and previous molecular investigations revealed a high diversity of fungi. In this study, we present a first trial to link fungal DNA-sequences with defined morphotypes characterized on the basis of partly new mycelial features obtained from field material of one tree species, Alzatea verticillata. Fine roots were halved lengthwise to study the mycelium anatomy on one half and to obtain fungal nuclear rDNA coding for the small subunit rRNA of Glomeromycota from the other half. Light microscopy revealed conspicuously large amounts of mycelium attaching to the surface of the rootlets. The mycelium formed fine- or large-branched appressoria-like plates, vesicles of regular or irregular shape, and very fine, multibranched structures ensheathed by septate hyphae. These previously undescribed features of the supraradical mycelia combined with intraradical mycelium structures were used for distinguishing of four main morphogroups and subordinate 14 morphotypes. DNA sequences of Glomus group A, Acaulospora and Gigaspora, were obtained and linked to three morphogroups. Two sequence types within Glomus group A could be tentatively associated to subordinate morphotypes.

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Figs. 1, 2 and 3 Fig. 1 Fig. 2 Fig. 3
Figs. 4, 5 and 6 Figs. 4a and b Fig. 5 Fig. 6
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Figs. 13 and 14 Fig. 13a and b Fig. 14
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Acknowledgements

The authors are indebted to the Deutsche Forschungsgemeinschaft for the generous financial support (DFG FOR 402). We are grateful to the NCI (former Fundacíon Científica San Francisco) for providing research facilities. We thank Tesfaye Wubet for providing primers and Michael Weiß for assistance in phylogenetic calculations.

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  1. Systematic Botany, Mycology and Botanical Garden, Eberhard-Karls-University Tübingen, Auf der Morgenstelle 1, 72076, Tübingen, Germany

    Adela Beck, Ingeborg Haug, Franz Oberwinkler & Ingrid Kottke

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  1. Adela Beck
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  2. Ingeborg Haug
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Correspondence to Adela Beck.

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Fig. S1

Scheme illustrating handling of fine root systems of A. verticillata for microscopy and parallel DNA extraction (GIF 32 kb)

High resolution image file (TIF 2455 kb)

Fig. S2

Overview and details of the supraradical mycelium of AM associated with A. verticillata. a Root surface with abundant supraradical mycelium (sm), b–f branched appressoria-like plates displaying different staining, g supraradical vesicles of irregular shape with differently staining layers, –k fine, multibranched structures ensheathed by brown septate hyphae (bh). Scale bars 10 μm (GIF 402 kb)

High resolution image file (TIF 17589 kb)

Fig. S3

a Fine, hyaline-staining besom-like branched (blb) appressoria-like plates, b fine, dark-staining besom-like branches, c wider branched appressoria-like plate displaying differently staining layers. d and e Supraradical mycelium of morphogroup IV displaying extremely large, multibranched structures (mb) not ensheated by brown septate hyphae; lobed supraradical vesicle (sv) and thick hyphae with differently staining wall layers (arrows). Scale bars 10 μm (GIF 339 kb)

High resolution image file (TIF 5026 kb)

Table S1

Characters used for distinguishing the AM morphotypes as presented in Tables 1, 2, and 3 and Figs. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 and 17 (DOC 41 kb)

Table S2

Primer names, sequences, and references (DOC 23 kb)

Table S3

Primer combinations, annealing temperatures used for nested PCR, and taxa identified (DOC 24 kb)

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Beck, A., Haug, I., Oberwinkler, F. et al. Structural characterization and molecular identification of arbuscular mycorrhiza morphotypes of Alzatea verticillata (Alzateaceae), a prominent tree in the tropical mountain rain forest of South Ecuador. Mycorrhiza 17, 607–625 (2007). https://doi.org/10.1007/s00572-007-0139-0

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