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Four mycelial strains of Entoloma clypeatum species complex form ectomycorrhiza-like roots with Pyrus betulifolia seedlings in vitro, and one develops fruiting bodies 2 months after inoculation

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Abstract

Entoloma clypeatum species complex (ECSC) forms ectomycorrhiza-like roots (EMLR) with host plant species of Rosaceae or Ulmaceae. The EMLR colonized with ECSC are characterized by a thick fungal mantle, absence of a Hartig net structure, and collapse of the apical meristem caused by hyphal invasion. Some researchers have suggested parasitism of ECSC because of this unique mode of colonization; however, the nature of the interaction between ECSC and host plants has not been investigated in co-culture because of the difficulty of culturing this group of fungi. We established a procedure to synthesize EMLR of ECSC on pear seedlings using fungal cultures. Three conspecific strains of ECSC isolated from basidiospores and one strain isolated from EMLR were tested. Cultured mycelia were inoculated onto a modified Norkrans’ C (MNC) or Hyponex-yeast-glucose (HYG) medium slant on the bottom of a polycarbonate jar and covered with autoclaved andosol or a vermiculite/sphagnum moss mixture (VSM); an axenically cultivated Pyrus betulifolia seedling was then planted in the jar. Five months after inoculation, the formation of EMLR with Hartig net-like hyphae was confirmed in all of the experimental plots. However, the rate of root colonization was significantly higher in experimental plots using andosol than in those using VSM. The growth of pear seedlings was similar irrespective of the level of root colonization, suggesting commensalism rather than parasitism of ECSC. One experimental plot using strain A3, an MNC slant, and andosol as a substrate produced ECSC fruiting bodies with mature basidia and basidiospores. The results suggested that our procedure enables the synthesis of EMLR of ECSC and cultivation of their fruiting bodies.

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Acknowledgements

We thank Dr. Akiyoshi Yamada for technical advice in our experiment. We thank Ms. Sachiko Ueta, Ms. Ayako Eriguchi, Ms. Mizuki Yokono, Ms. Masako Oka, and Ms. Kiko Hirata for cryopreservation of ECSC strains. We thank Ms. Tatsumi Matsushima and Mr. Tomio Fuji for their assistance in sampling ECSC specimens at the farm of Ms. Matsushima, Yurihama-cho, Tottori Prefecture. We also thank the staff of the Division of Instrumental Research, Research Center for Supports to Advanced Science, Shinshu University, for their technical support regarding DNA sequencing.

Funding

This study was supported in part by a Grant-in-Aid for JSPS KAKENHI Grant Number JP18K14458 from the Ministry of Education, Culture, Sports, Science and Technology, Japan. This study was also supported by a research fund from the president of Tottori University.

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Authors and Affiliations

  1. Graduate School of Sustainable Science, Tottori University, 4-101 Koyama, Tottori, 680-8553, Japan

    Manami Shishikura

  2. Faculty of Agriculture, Tottori University, 4-101 Koyama, Tottori, 680-8553, Japan

    Yoshihiro Takemura

  3. Fungus/Mushroom Resource and Research Center, Faculty of Agriculture, Tottori University, 4-101 Koyama, Tottori, 680-8553, Japan

    Kozue Sotome, Nitaro Maekawa, Akira Nakagiri & Naoki Endo

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  1. Manami Shishikura
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  2. Yoshihiro Takemura
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  3. Kozue Sotome
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  4. Nitaro Maekawa
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  5. Akira Nakagiri
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  6. Naoki Endo
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Correspondence to Naoki Endo.

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All the experiments undertaken in this study comply with the current laws of the country where they were performed.

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Shishikura, M., Takemura, Y., Sotome, K. et al. Four mycelial strains of Entoloma clypeatum species complex form ectomycorrhiza-like roots with Pyrus betulifolia seedlings in vitro, and one develops fruiting bodies 2 months after inoculation. Mycorrhiza 31, 31–42 (2021). https://doi.org/10.1007/s00572-020-00994-4

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