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Characterization of mycorrhizal fungi isolated from the threatened Cypripedium macranthos in a northern island of Japan: two phylogenetically distinct fungi associated with the orchid

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An Erratum to this article was published on 11 August 2009

Abstract

We isolated Rhizoctonia-like fungi from populations of the threatened orchid Cypripedium macranthos. In ultrastructural observations of the septa, the isolates had a flattened imperforate parenthesome consisting of two electron-dense membranes bordered by an internal electron-lucent zone, identical to the septal ultrastructure of Rhizoctonia repens (teleomorph Tulasnella), a mycorrhizal fungus of many orchid species. However, hyphae of the isolates did not fuse with those of known tester strains of R. repens and grew less than half as fast as those of R. repens. In phylogenetic analyses, sequences for rDNA and internal transcribed spacer (ITS) regions of the isolates were distinct from those of the taxonomically identified species of Tulasnella. On the basis of the ITS sequences, the isolates clustered into two groups that corresponded exactly with the clades demonstrated for other Cypripedium spp. from Eurasia and North America despite the geographical separation, suggesting high specificity in the Cypripedium–fungus association. In addition, the two phylogenetic groups corresponded to two different plant clones at different developmental stages. The fungi from one clone constituted one group and did not belong to the other fungal group isolated from the other clone. The possibility of switching to a new mycorrhizal partner during the orchid’s lifetime is discussed.

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Acknowledgments

We are most grateful to Dr. Chikara Masuta for many helpful discussions and suggestions. We thank Mr. Akihiko Matsuzawa for his generous help in molecular works. This work was supported in part by a Grant-in-Aid (Conservation study on designated national endangered plants with the model case C. macranthos var. rebunense) from the Ministry of the Environment, Japan, and by a Grant-in-Aid (no. 16310157) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Laboratory of Crop Physiology, Graduate School of Agriculture, Hokkaido University, Kita-ku, Kita 9, Nishi 9, Sapporo, 060-8589, Japan

    Hanako Shimura, Mayumi Matsuura & Yasunori Koda

  2. Laboratory of Plant Pathology, Graduate School of Agriculture, Hokkaido University, Sapporo, 060-8589, Japan

    Mai Sadamoto & Shigeo Naito

  3. Forest Dynamics and Diversity Group, Hokkaido Research Center, Forestry and Forest Products Research Institute, Sapporo, 062-8516, Japan

    Takayuki Kawahara

Authors
  1. Hanako Shimura
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  2. Mai Sadamoto
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  3. Mayumi Matsuura
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  4. Takayuki Kawahara
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  5. Shigeo Naito
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  6. Yasunori Koda
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Corresponding author

Correspondence to Hanako Shimura.

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Supplementary Fig. S1

Phylogenetic tree of the ITS–5.8S sequences from fungal isolates from Cypripedium on Rebun island and Epulorhiza spp. (teleomorph Tulasnella spp.) isolates. The best tree resulting from heuristic maximum likelihood analysis in PAUP* is presented with support values derived using 1,000 bootstrap ML replicates (values >70% are shown). Cypripedium isolates were boxed. Unidentified mycorrhizal fungi of Cypripedium species collected worldwide (bold face; Shefferson et al. 2007) were also included in this analysis to compare with the fungi we isolated. Accession DQ925516 and DQ925552 were derived from C. macranthos var. rebunense, accession DQ925643 was derived from C. macranthos var. speciosum, and other accessions were derived from other Cypripedium spp. from Eurasia and North America (Shefferson et al. 2007). Athelia (Sclerotium) rolfsii (AY684917) was used as an outgroup (Sharon et al. 2008). The isolates from Cypripedium grouped in separate clusters, distant from the clusters of the Tulasnella and Epulorhiza groups. The clade TG-A to TG-F are classified by Sharon et al. (2008) (PPT 1594 kb)

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Shimura, H., Sadamoto, M., Matsuura, M. et al. Characterization of mycorrhizal fungi isolated from the threatened Cypripedium macranthos in a northern island of Japan: two phylogenetically distinct fungi associated with the orchid. Mycorrhiza 19, 525–534 (2009). https://doi.org/10.1007/s00572-009-0251-4

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