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Estimation of the most suitable nitrogen concentration for sporocarp formation in Laccaria japonica colonizing Pinus densiflora seedlings through in vitro mycelial culture

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Abstract

Many ectomycorrhizal (ECM) fungi produce commercially valuable edible sporocarps. However, the effects of nitrogen (N) application on ECM fungal sporocarp formation remain poorly understood. In this study, we investigated the effect of application of various N concentrations (0, 5, 25, 50, 100, and 200 mg/L) on the growth of Laccaria japonica mycelia in vitro for 1 month. The results showed that L. japonica mycelial biomass was highest in the 50 mg/L treatment and was significantly inhibited at N concentrations higher than 200 mg/L. Next, we investigated the effects of N application on mycorrhizal colonization and sporocarp formation in L. japonica colonizing Pinus densiflora seedlings in pots. The seedlings were watered with nutrient solutions containing 0, 5, 25, 50, or 100 mg N/L. The biomass, photosynthetic rate, and mycorrhizal colonization rates of the seedlings were measured at 45 days (first appearance of primordia), 65 days (sporocarp appearance on the substrate surface), and 4 months after seedlings were transplanted. The numbers of primordia and sporocarps were recorded during the experimental period. Total carbon (C) and N content were determined in seedlings at 4 months after transplantation, and in L. japonica sporocarps. Both mycelial growth and sporocarp production reached their maximum at an N application concentration of 50 mg/L, suggesting that the most suitable N concentration for ECM fungal sporocarp formation can easily be estimated in vitro during mycelial growth. This finding may help determine the most suitable N conditions for increasing edible ECM fungus sporocarp production in natural forests.

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Acknowledgements

We are grateful to Mitsuko Goto, Yang Liu, Suguru Tanaka, Masayuki Kubota, Huayong Wang for technical assistance, the staff of the University of Tokyo Tanashi Forest for providing the forest soils, Kazuhide Nara for providing the culture of ectomycorrhizal fungus and Maki Narimatsu for providing the seeds of P. densiflora. We thank two anonymous reviewers for revising and useful comments on the manuscript.

Funding

This work was supported in part by JSPS KAKENHI (17H03824).

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Authors and Affiliations

  1. Laboratory of Forest Symbiology, Asian Research Center for Bioresource and Environmental Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

    Shijie Zhang, Momi Tsuruta, Chaofeng Li, Ruiyang Xu, Jiali Li & Chunlan Lian

  2. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, China

    Shijie Zhang

  3. Department of Forest Sciences, University of Helsinki, PO Box 27, 00014, Helsinki, Finland

    Lu-Min Vaario

  4. College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China

    Yan Xia

  5. Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, Japan

    Norihisa Matsushita & Hiroyuki Kurokochi

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  1. Shijie Zhang
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Zhang, S., Tsuruta, M., Li, C. et al. Estimation of the most suitable nitrogen concentration for sporocarp formation in Laccaria japonica colonizing Pinus densiflora seedlings through in vitro mycelial culture. Mycorrhiza 32, 451–464 (2022). https://doi.org/10.1007/s00572-022-01085-2

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