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Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ

Received: 26 July 2023     Accepted: 4 September 2023     Published: 18 September 2023
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Abstract

We studied on the ectomycorrhiza formation of Tricholoma matsutake on mature pine (Pinus densiflora) root using matsutake solid spawn in situ. To find out suitable medium for the solid spawn, the matsutake mycelium growths on different media were compared. Among them, GYP medium (glucose 20g, yeast extract 5g, peptone 5g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) and GWR medium (glucose 20g, wheat bran 50g, rice bran 50g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) showed good proliferation of matsutake mycelium. The GYP and GWR media were used to make charcoal and vermiculite-soil spawns. The ergosterol amounts of charcoal and vermiculite-soil spawns were 1.53 and 1.61 times higher than that of the natural shiro, respectively. In spring, the roots of about 30-year-old red pine were shortened with unsterilized scissors. About twelve months later, the regenerated root tips from shortened points were inoculated with charcoal and vermiculite-soil spawns, respectively. After four months by the inoculation with charcoal and vermiculite-soil spawns, 55% and 50% of experimental plots inoculated with charcoal and vermiculite-soil spawns revealed dichotomic root tips characteristic to ectomycorrhiza, respectively. The nested PCR amplification results confirmed that the dichotomous root tips were colonised by matsutake spawn. Our findings demonstrated that the ectomycorrhiza of T. matsutake could be formed on the mature pine root in situ by using matsutake solid spawns.

Published in International Journal of Ecotoxicology and Ecobiology (Volume 8, Issue 3)
DOI 10.11648/j.ijee.20230803.13
Page(s) 41-48
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2023. Published by Science Publishing Group

Keywords

Tricholoma matsutake, Ectomycorrhiza, Solid Spawn, Inoculation, Pinus desiflora, Nested PCR

References
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[2] ENDO N., DOKMAI P., SUWANNASAI N., PHOSRI C., HORIMAI Y., HIRAI N., et al. 2015. Ectomycorrhization of Tricholoma matsutake with Abies veitchii AND Tsuga diversifolia in the subalpine forests of Japan. Mycoscience. 56: 402-412.
[3] VAARIO L. M., SAVONEN E. M., PELTONIEMI M., MIYAZAWA T., PULKKINEN P., AND SARJALAT. 2015. Fruiting pattern of Tricholoma matsutake in southern Finland. Scandinavian Journal of Forest Research. 30: 259~265.
[4] OHARA H., AND HAMADA M. 1967. Disappearance of bacteria from the zone of active mycorrhizas in Tricholoma matsutake (S. Ito et Imai) singer. Nature. 213: 528-529.
[5] NARIMATSU M., KOIWA T., MASAKI T., SAKAMOTO Y., OHMORI H., AND TAWARAYA K. 2015. Relationship between climate, expansion rate, and fruiting in fairy rings (‘shiro’) of an ectomycorrhizal fungus Tricholoma matsutake in a Pinus densifloraforest. Fungal Ecology. 15: 18~28.
[6] TAKASHI Y., AKIYOSHI Y., AND HITOSHI F. 2020. Advances in the cultivation of the highly-prized ectomycorrhizal mushroom Tricholoma matsutake. Mycoscience. 61: 49-57.
[7] KUSUDA M., UEDA M., MIYATAKE K., AND TERASHITA T. 2008. Characterization of the carbohydrase productions of an ectomycorrhizal fungus, Tricholoma matsutake. Mycoscience. 49: 291-297. https://doi.org/10.1007/S10267-008-0423-7.
[8] KUSUDA M., UEDA M., NAKAZAWA M., MIYATAKE K., AND YAMANAKA K. 2006. Detection of b-glucosidase as saprotrophic ability from an ectomycorrhizal mushroom, Tricholoma matsutake. Mycoscience. 47: 184e189.
[9] SHIMOKAWA T., YAMAGUCHI M., AND MURATA H. 2017. Agar plate assays using dye linked substrates differentiate members of Tricholoma sect. caligata, ectomycorrhizal symbionts represented by Tricholoma matsutake. Mycoscience. 58: 432-437.
[10] YAMADA A., MAEDA K., AND OHMASA M. 1999a. Ectomycorrhiza formation of Tricholoma matsutake isolates on seedlings of Pinus densiflora in vitro. Mycoscience. 40: 455-463.
[11] GUERIN-LAGUETTE A., VAARIO L. M., GILLW M., LAPEYRIE F., MATSUSHITA N., AND SUZUKI K. 2000. Rapid in vitro ectomycorrhizal infection on Pinus densiflora roots by Tricholoma matsutake. Mycoscience. 41: 389-393.
[12] VAARIO L. M., GUERIN-LAGUETTE A., GILL W. M., LAPEYRIE F., AND 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. Journal of Forest Research. 5: 293-297.
[13] YAMADA A., KANEKAWA S., AND OHMASA M. 1999b. Ectomycorrhiza formation of Tricholoma matsutake on Pinus densiflora. Mycoscience. 40: 193-198. https://doi.org/10.1007/BF02464298.
[14] GUERIN-LAGUETTE A., VAARIO L. M., MATSUSHITA N., SHINDO K., SUZUKI K., AND LAPEYRIE F. 2003. Growth stimulation of a Shiro-like, mycorrhiza forming, mycelium of Tricholoma matsutake on solid substrates by non-ionic surfactants or vegetable oil. Mycological Progress. 2: 37-44.
[15] KA K. H., KIM H. S., HUR T. C., PARK H., JEON S. M., RYOO R., et al. 2018. Analysis of environment and production of Tricholoma matsutake in matsutake-infected pine trees (in Korean). Korean Journal of Mycology. 46: 34-42.
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[18] LEE W. H., HAN S. K., KIM B. S., BHUSHAN S, LEES Y., KOC S., SUNG G. H., AND SUNG J. M. 2007. Proliferation of Tricholoma matsutake Mycelial Mats in Pine Forest Using Mass Liquid Inoculum. Mycobiology. 35 (2): 54-61.
[19] HAMADA M. 1964. Method of pure culture of the mycelium of Armillaria matsutake and of its allies. In: The Matsutake Research Association (Ed.), Matsutake (Tricholoma matsutake Singer)-Its fundamental studies and economic production of the fruit-body (pp. 97-100). Kyoto: Nakanishi printing (in Japanese).
[20] KIKUCHI K., MATSUSHITA N., GUERIN-LAGUETTE A., OHTA A. AND SUZUKI K. 2000. Detection of Tricholoma matsutakeby specific primers. Mycol. Res. 104: 1427-1430.
[21] ZHAO X. R., LINA Q., AND BROOKES P. C. 2005. Does soil ergosterol concentration provide a reliable estimate of soil fungal biomass? Soil biology and biochemistry. 37: 311-315.
[22] WHITE T. J., BRUNS T., LEE S., AND TAYLOR J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. P 315–322 in Innis MA, Gelfand DH, Snisky JJ, White TJ (eds) PCR protocols. A guide to methods and applications. Academic, San Diego, CA.
[23] Kawai M., IMAJI A., YAMADA A., AND KINOSHITA A. 2018. Shiro formation and fruit body flashing by inoculation of Tricholoma bakamatsutake mycelia in the forest. P. 25 in Abstracts of the annual meeting of the mycological Society of Japan. https://doi.org/10.11556/msj7abst.62.0_25.
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    Yong Chan Kim, Kol Nam, Myong Song Kim, Un Chol Kim, Un Hyok Kim, et al. (2023). Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ. International Journal of Ecotoxicology and Ecobiology, 8(3), 41-48. https://doi.org/10.11648/j.ijee.20230803.13

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    ACS Style

    Yong Chan Kim; Kol Nam; Myong Song Kim; Un Chol Kim; Un Hyok Kim, et al. Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ. Int. J. Ecotoxicol. Ecobiol. 2023, 8(3), 41-48. doi: 10.11648/j.ijee.20230803.13

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    AMA Style

    Yong Chan Kim, Kol Nam, Myong Song Kim, Un Chol Kim, Un Hyok Kim, et al. Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ. Int J Ecotoxicol Ecobiol. 2023;8(3):41-48. doi: 10.11648/j.ijee.20230803.13

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  • @article{10.11648/j.ijee.20230803.13,
      author = {Yong Chan Kim and Kol Nam and Myong Song Kim and Un Chol Kim and Un Hyok Kim and Myong Bok Kim and Gum Chol Pak and Hui Won Kim},
      title = {Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ},
      journal = {International Journal of Ecotoxicology and Ecobiology},
      volume = {8},
      number = {3},
      pages = {41-48},
      doi = {10.11648/j.ijee.20230803.13},
      url = {https://doi.org/10.11648/j.ijee.20230803.13},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijee.20230803.13},
      abstract = {We studied on the ectomycorrhiza formation of Tricholoma matsutake on mature pine (Pinus densiflora) root using matsutake solid spawn in situ. To find out suitable medium for the solid spawn, the matsutake mycelium growths on different media were compared. Among them, GYP medium (glucose 20g, yeast extract 5g, peptone 5g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) and GWR medium (glucose 20g, wheat bran 50g, rice bran 50g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) showed good proliferation of matsutake mycelium. The GYP and GWR media were used to make charcoal and vermiculite-soil spawns. The ergosterol amounts of charcoal and vermiculite-soil spawns were 1.53 and 1.61 times higher than that of the natural shiro, respectively. In spring, the roots of about 30-year-old red pine were shortened with unsterilized scissors. About twelve months later, the regenerated root tips from shortened points were inoculated with charcoal and vermiculite-soil spawns, respectively. After four months by the inoculation with charcoal and vermiculite-soil spawns, 55% and 50% of experimental plots inoculated with charcoal and vermiculite-soil spawns revealed dichotomic root tips characteristic to ectomycorrhiza, respectively. The nested PCR amplification results confirmed that the dichotomous root tips were colonised by matsutake spawn. Our findings demonstrated that the ectomycorrhiza of T. matsutake could be formed on the mature pine root in situ by using matsutake solid spawns.},
     year = {2023}
    }
    

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  • TY  - JOUR
    T1  - Ectomycorrhiza Formation of Tricholoma matsutake on Mature Pine Root in Situ
    AU  - Yong Chan Kim
    AU  - Kol Nam
    AU  - Myong Song Kim
    AU  - Un Chol Kim
    AU  - Un Hyok Kim
    AU  - Myong Bok Kim
    AU  - Gum Chol Pak
    AU  - Hui Won Kim
    Y1  - 2023/09/18
    PY  - 2023
    N1  - https://doi.org/10.11648/j.ijee.20230803.13
    DO  - 10.11648/j.ijee.20230803.13
    T2  - International Journal of Ecotoxicology and Ecobiology
    JF  - International Journal of Ecotoxicology and Ecobiology
    JO  - International Journal of Ecotoxicology and Ecobiology
    SP  - 41
    EP  - 48
    PB  - Science Publishing Group
    SN  - 2575-1735
    UR  - https://doi.org/10.11648/j.ijee.20230803.13
    AB  - We studied on the ectomycorrhiza formation of Tricholoma matsutake on mature pine (Pinus densiflora) root using matsutake solid spawn in situ. To find out suitable medium for the solid spawn, the matsutake mycelium growths on different media were compared. Among them, GYP medium (glucose 20g, yeast extract 5g, peptone 5g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) and GWR medium (glucose 20g, wheat bran 50g, rice bran 50g, KH2PO4 1g, CaCl2 50mg and vitamin B1 50mg) showed good proliferation of matsutake mycelium. The GYP and GWR media were used to make charcoal and vermiculite-soil spawns. The ergosterol amounts of charcoal and vermiculite-soil spawns were 1.53 and 1.61 times higher than that of the natural shiro, respectively. In spring, the roots of about 30-year-old red pine were shortened with unsterilized scissors. About twelve months later, the regenerated root tips from shortened points were inoculated with charcoal and vermiculite-soil spawns, respectively. After four months by the inoculation with charcoal and vermiculite-soil spawns, 55% and 50% of experimental plots inoculated with charcoal and vermiculite-soil spawns revealed dichotomic root tips characteristic to ectomycorrhiza, respectively. The nested PCR amplification results confirmed that the dichotomous root tips were colonised by matsutake spawn. Our findings demonstrated that the ectomycorrhiza of T. matsutake could be formed on the mature pine root in situ by using matsutake solid spawns.
    VL  - 8
    IS  - 3
    ER  - 

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Author Information
  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Laboratory of Matsutake, Central Institute of Mushroom, Agricultural Committee, Pyongyang, Korea

  • Department of Daily Food, Institute of Microbiology, The State Academy of Sciences, Pyongyang, Korea

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