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Spore germination and ectomycorrhizae formation of Tricholoma matsutake on pine root systems with previously established ectomycorrhizae from a dikaryotic mycelial isolate of T. matsutake

Mycorrhiza volume 31, pages 335–347 (2021)Cite this article

Abstract

In vitro ectomycorrhizal synthesis of Tricholoma matsutake with host plants has been widely conducted to elucidate fungal symbiotic properties for future cultivation practices. Here, we report on the importance of basidiospore inocula for this fungus to provide ectomycorrhizal seedlings in vitro. Ectomycorrhizal pine seedlings synthesized in vitro with cultured mycelium of T. matsutake (isolate #45 or #84) in a 250-mL culture vessel (soil volume) were transplanted to a large 1-L culture vessel. Fresh basidiospores of this fungus were aseptically inoculated on the ectomycorrhizal root system. The ectomycorrhizal seedlings in the 1-L vessel were grown for 9 months, and some plants were further grown for 6 more months under non-aseptic conditions in 4.1-L jars. The ectomycorrhizal seedlings previously inoculated with isolate #84 in the 1-L vessel showed significant ectomycorrhizal biomass (mycorrhizal root length) after spore inoculation. The ectomycorrhizal seedlings in the 4.1-L vessel showed large shiro structures (> 10 cm in diameter). PCR amplification of intergenic spacer 1 of the rRNA gene and long terminal repeat retroelement of T. matsutake in ectomycorrhizal root tips in both the 1-L vessels and 4.1-L jars revealed the presence of amplicons of the previously inoculated culture isolate of T. matsutake and the new genet(s) that established via germination of the inoculated basidiospores. This is the first report that inoculated basidiospores of T. matsutake germinated and colonized the host root to generate ectomycorrhizae in vitro.

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Acknowledgements

We acknowledge Masamichi Ichikawa for the supply of fresh Tricholoma matsutake samples in the spore inoculation experiment, and Hitoshi Murata in Forestry and Forest Products Research Institute for the technical advice in DNA analyses. We thank the members of Applied Mycology Laboratory, Shinshu University for their support of this study.

Funding

This study was supported in part by KAKENHI Grant Number 15H01751 from the Japan Society for the Promotion of Science (JSPS), and a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan, “Technology development for the optimal use of forest resources.”

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Authors and Affiliations

  1. Department of Agriculture, Graduate School of Science and Technology, Shinshu University, Nagano, Minami-minowa, 8304399-4598, Japan

    Yuka Horimai, Hiroki Misawa, Yu Tateishi, Masaki Fukuda & Akiyoshi Yamada

  2. Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Nagano, Minami-minowa, 8304399-4598, Japan

    Kentaro Suzuki, Yu Tateishi, Masaki Fukuda & Akiyoshi Yamada

  3. Nagano Prefectural Forest Research Center, Kataoka, Shiojiri, Nagano, 399-0711, Japan

    Hitoshi Furukawa

  4. Forestry and Forest Products Research Institute, Matsunosato, Tsukuba, Ibaraki, 305-8687, Japan

    Takashi Yamanaka

  5. KOA Corporation, Nagano, Nakaminowa, 14016399-4697, Japan

    Shozo Yamashita & Toshiharu Takayama

  6. Division of Mountain Ecosystem, Institute for Mountain Science, Shinshu University, Nagano, Minami-minowa, 8304399-4598, Japan

    Akiyoshi Yamada

Authors
  1. Yuka Horimai
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  2. Hiroki Misawa
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  3. Kentaro Suzuki
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  4. Yu Tateishi
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  7. Shozo Yamashita
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  8. Toshiharu Takayama
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  10. Akiyoshi Yamada
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Correspondence to Akiyoshi Yamada.

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Horimai, Y., Misawa, H., Suzuki, K. et al. Spore germination and ectomycorrhizae formation of Tricholoma matsutake on pine root systems with previously established ectomycorrhizae from a dikaryotic mycelial isolate of T. matsutake. Mycorrhiza 31, 335–347 (2021). https://doi.org/10.1007/s00572-021-01028-3

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Keywords

  • Edible mycorrhizal mushrooms
  • Fungus–plant interaction
  • Genetic diversity
  • Pinus densiflora
  • Seedling growth