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Successful inoculation of mature pine with Tricholoma matsutake

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Abstract

Our finding demonstrates, for the first time, that the roots of mature pine trees can be successfully inoculated with a symbiotic ectomycorrhizal fungus, the valuable matsutake mushroom. Long root segments (ca. 5–10 mm in diameter, ca. 50 cm in length) of 50-year-old Pinus densiflora trees were excavated, washed, auxin-treated (2–5 mg indole butyric acid, IBA, per root) and incubated in moist Spagnum moss. Twelve months later, short roots were regenerated, of which approximately 90% were free of mycorrhizae. Mycorrhiza-free short roots were inoculated with mycelial pieces of Tricholoma matsutake and incubated further in a sterilized substrate. Four-and-a-half months later, roots putatively colonized by Matsutake were sampled near the inoculation points. A T. matsutake-specific ITS-rDNA fragment was amplified by nested PCR from approximately 80% of the root samples analyzed, whereas approximately 66% of the root samples processed for staining with Chlorazol black E displayed characteristic T. matsutake Hartig net structures. These results confirm the symbiotic infection of mature P. densiflora roots by matsutake.

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References

  • Gill WM, Lapeyrie F, Gomi T, Suzuki K (1999) Tricholoma matsutake—an assessment of in situ and in vitro infection by observing cleared and stained whole roots. Mycorrhiza 9:227–231

    Google Scholar 

  • Gill WM, Guerin-Laguette A, Lapeyrie F, Suzuki K (2000) Matsutake—morphological evidence of ectomycorrhiza formation between Tricholoma matsutake and host roots in a pure Pinus densiflora forest stand. New Phytol 147:381–388

    Google Scholar 

  • Guerin-Laguette A, Vaario LM, Gill WM, Lapeyrie F, Matsushita N, Suzuki K (2000a) Rapid in vitro ectomycorrhizal infection on Pinus densiflora roots by Tricholoma matsutake. Mycoscience 41:389–393

    Google Scholar 

  • Guerin-Laguette A, Plassard C, Mousain D (2000b) Effects of experimental conditions on mycorrhizal relationships between Pinus sylvestris and Lactarius deliciosus and unprecedented fruit-body formation of the Saffron milk cap under controlled soilless conditions. Can J Microbiol 46:790–799

    Google Scholar 

  • Guerin-Laguette A, Matsushita N, Kikuchi K, Iwase K, Lapeyrie F, Suzuki K (2002) Identification of a prevalent Tricholoma matsutake ribotype in Japan by rDNA IGS1 spacer characterization. Mycol Res 106:435–443

    Google Scholar 

  • Guerin-Laguette A, Vaario LM, Matsushita N, Shindo K, Suzuki K, Lapeyrie F (2003) Growth stimulation of Tricholoma matsutake on solid substrates by non-ionic surfactants or vegetable oil. Mycol Prog 2:37–44

    Google Scholar 

  • Guerin-Laguette A, Shindo K, Matsushita N, Suzuki K, Lapeyrie F (2004) The mycorrhizal fungus Tricholoma matsutake stimulates Pinus densiflora seedling growth in vitro. Mycorrhiza 14:397–400

    Google Scholar 

  • Hall IR, Wang Y, Amicucci A (2003) Cultivation of edible ectomycorrhizal mushrooms. Trends Biotechnol 21:433–438

    Google Scholar 

  • Hosford D, Pilz D, Molina R, Amaranthus M (1997) Ecology and management of the commercially harvested American matsutake. USDA general technical report PNW-GTR-412

  • Ito T (2000) In: Ogawa M, Kinugawa K (eds) Mushroom handbook. Asakura, Japan, pp 9–21

    Google Scholar 

  • Kikuchi K, Matsushita N, Guerin-Laguette A, Ohta A, Suzuki K (2000) Detection of Tricholoma matsutake by specific ITS primers. Mycol Res 104:1427–1430

    Google Scholar 

  • Kraigher H, Agerer R, Javornik B (1995) Ectomycorrhizae of Lactarius lignyotus on Norway spruce, characterized by anatomical and molecular tools. Mycorrhiza 5:175–180

    Google Scholar 

  • Ogawa M (1982) How to produce matsutake mushroom. Matsutake Research and Consultation Office, Forest Service Research Station, Nara, Japan

  • Ohta A (1990) A new medium for mycelial growth of mycorrhizal fungi. Trans Mycol Soc Japan 31:323–334

    Google Scholar 

  • Suzuki K (2005) Ectomycorrhizal ecophysiology and the puzzle of Tricholoma matsutake. J Jpn For Soc 87:90–102 (in Japanese with English summary)

    Google Scholar 

  • Vaario LM, Guerin-Laguette A, Gill WM, Lapeyrie F, Suzuki K (2000) Only two weeks are required for Tricholoma matsutake to differentiate ectomycorrhizal Hartig net structures in roots of Pinus densiflora seedlings cultivated on artificial substrate. J For Res 5:293–297

    Google Scholar 

  • Vaario LM, Guerin-Laguette A, Suzuki K, Lapeyrie F (2002) Investigation of Tricholoma matsutake saprotrophic potential: growth over pine bark treated with surfactants. Mycorrhiza 12:1–5

    Google Scholar 

  • Wang Y (1995) Tricholoma matsutake. PhD thesis, University of Otago, Dunedin, New Zealand. 3 volumes

  • Wang Y, Hall IR (2004) Edible ectomycorrhizal mushrooms: challenges and achievements. Can J Bot 82:1063–1073

    Google Scholar 

  • Wang Y, Hall IR, Evans LA (1997) Ectomycorrhizal fungi with edible fruiting bodies. 1. Tricholoma matsutake and related fungi. Econ Bot 51:311–327

    Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Snisky JJ, White TJ (eds) PCR protocols. A guide to methods and applications. Academic, San Diego, CA, pp 315–322

    Google Scholar 

  • Yamada A, Kanekawa S, Ohmasa M (1999a) Ectomycorrhiza formation of Tricholoma matsutake on Pinus densiflora. Mycoscience 40:193–198

    Google Scholar 

  • Yamada A, Maeda K, Ohmasa M (1999b) Ectomycorrhiza formation of Tricholoma matsutake isolates on seedlings of Pinus densiflora in vitro. Mycoscience 40:455–463

    Google Scholar 

  • Yoshimura F (2004) This is how far we’ve got! Matsutake cultivation. Toronto, 109 pp (in Japanese)

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Acknowledgements

This research was supported by the Japanese Society for the Promotion of Science (JSPS) Project No. 14104006.

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Correspondence to Alexis Guerin-Laguette.

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Guerin-Laguette, A., Matsushita, N., Lapeyrie, F. et al. Successful inoculation of mature pine with Tricholoma matsutake. Mycorrhiza 15, 301–305 (2005). https://doi.org/10.1007/s00572-005-0355-4

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  • DOI: https://doi.org/10.1007/s00572-005-0355-4

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