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The effects of co-colonising ectomycorrhizal fungi on mycorrhizal colonisation and sporocarp formation in Laccaria japonica colonising seedlings of Pinus densiflora

  • Shijie Zhang
  • Lu-Min Vaario
  • Yan Xia
  • Norihisa Matsushita
  • Qifang Geng
  • Momi Tsuruta
  • Hiroyuki Kurokochi
  • Chunlan LianEmail author
Original Article


Forest trees are colonised by different species of ectomycorrhizal (ECM) fungi that interact competitively or mutualistically with one another. Most ECM fungi can produce sporocarps. To date, the effects of co-colonising fungal species on sporocarp formation in ECM fungi remain unknown. In this study, we examined host plant growth, mycorrhizal colonisation, and sporocarp formation when roots of Pinus densiflora are colonised by Laccaria japonica and three other ECM fungal species (Cenococcum geophilum, Pisolithus sp., and Suillus luteus). Sporocarp numbers were recorded throughout the experimental period. The biomass, photosynthetic rate, and mycorrhizal colonisation rate of the seedlings were also measured at 45 days, 62 days, and 1 year after seedlings were transplanted. Results indicated that C. geophilum and S. luteus may negatively impact mycorrhizal colonisation and sporocarp formation in L. japonica. Sporocarp formation in L. japonica was positively correlated with conspecific mycorrhizal colonisation but negatively correlated with the biomass of seedlings of P. densiflora. The co-occurring ECM fungi largely competed with L. japonica, resulting in various effects on mycorrhizal colonisation and sporocarp formation in L. japonica. A variety of mechanisms may be involved in the competitive interactions among the different ECM fungal species, including abilities to more rapidly colonise root tips, acquire soil nutrients, or produce antibiotics. These mechanisms need to be confirmed in further studies.


Sporocarps formation Co-colonisation Ectomycorrhizal fungi Laccaria japonica Pinus densiflora 



We are grateful to Mitsuko Goto, Ruiyang Xu, Jiali Li, Suguru Tanaka, Masayuki Kubota for technical assistance; the staff of the University of Tokyo Tanashi Forest for providing the forest soils and the vacuum freeze dryer; Kazuhide Nara for providing the cultures of ECM fungi; Maki Narimatsu for providing the seeds of P. densiflora. We are thankful to Kenji Fukuda for providing insightful comments which improved this manuscript. We thank Dr. Jan Colpaert and two anonymous reviewers for revising and useful comments on the manuscript.

Funding information

This work was supported in part by a JSPS KAKENHI (granted to CL, no. 17H03824).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

572_2019_890_MOESM1_ESM.pdf (42 kb)
Supplementary Figure S1 Agarose gel electrophoresis of PCR products from amplification using the primer Lj03f/03r. Lane M: 100 bp ladder marker. Lane 1: DNA extracted from mycelia of Laccaria japonica. Lane 2: negative control. Lane 3: DNA extracted from mycelia of Cenococcum geophilum. Lane 4: DNA extracted from mycelia of Pisolithus sp. Lane 5: DNA extracted from mycelia of Suillus luteus. Lane 6: DNA extracted from soil control. Lane 7: DNA extracted from non-mycorrhizal root tips. (PDF 42 kb)
572_2019_890_MOESM2_ESM.pdf (31 kb)
Supplementary Figure S2 The net photosynthetic rates of (a) non-mycorrhizal seedlings (NM), (b) seedlings colonised by Laccaria japonica (Lj), and (c) seedlings colonised by Cenococcum geophilum (Cg), Pisolithus sp. (PS), and Suillus luteus (Sl) at 45 days, 62 days, and 1 year after seedlings were transplanted into pots. Each pot contained four seedlings. Con: control with only NM seedlings; Lj + NM: treatment with NM seedlings and Lj-colonised seedlings; Lj + Cg: treatment with NM seedlings and Lj- and Cg-colonised seedlings; Lj + PS: treatment with NM seedlings and Lj- and PS-colonised seedlings; Lj + Sl: treatment with NM seedlings and Lj- and Sl-colonised seedlings. Values are represented as mean ± SD (n = 3–5). Pairwise comparisons using the Wilcoxon rank sum test were conducted to test for differences among treatments. Letters indicate significant differences (P < 0.05, P value adjusted using the BH method). The Steel test was also used to test for differences between the control and each treatment, *P < 0.05. (PDF 30 kb)
572_2019_890_MOESM3_ESM.docx (17 kb)
Supplementary Table S1 (DOCX 16 kb)
572_2019_890_MOESM4_ESM.docx (16 kb)
Supplementary Table S2 (DOCX 16 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Shijie Zhang
    • 1
  • Lu-Min Vaario
    • 2
    • 3
  • Yan Xia
    • 4
  • Norihisa Matsushita
    • 2
  • Qifang Geng
    • 1
  • Momi Tsuruta
    • 1
  • Hiroyuki Kurokochi
    • 2
  • Chunlan Lian
    • 1
    Email author
  1. 1.Asian Natural Environmental Science CenterThe University of TokyoTokyoJapan
  2. 2.Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  3. 3.Department of Forest SciencesUniversity of HelsinkiHelsinkiFinland
  4. 4.College of Life SciencesNanjing Agricultural UniversityNanjingChina

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