Table S1. Macro- and microscopic features of ectomycorrhizal phylotypes on current-year or older Pinus thunbergii seedlings in a Japanese coastal forest.

Phylotype / Color / Mantle
structurea / Clamp
connection / Cystidia / Emanatinghyphae (µm) / Other features / Seedling ages
Current / Older b
Basidiomycete sp.1 / White / FP / ○ / 2–3 / ○ / ○
Lactarius quieticolor / White / NS / - / Laticiferouscells / ○ / ○
Clavulinaceae sp.1 / White / NS (-NIS) / 1–3 / Lipid materials, rhizomorph (10 to 20 µm) / ○ / ○
Thelephoraceae sp.1 / White / NIS / ○ / 2–5 / ○ / ○
Russula sp. 1 / White / IIS / - / Gelatinous substances / ○ / ○
Trichophaea sp.1 / Brown / FP / ○ / 1–3 / ○ / ○
Sebacinaceae sp.1 / Brown / NP / 1–2 / ○ / ○
Bankeraceae sp.1 / Brown / NP / 1–3 / Crystal substances / ○
Tomentellaceae sp.1 / Brown / NS(-NIS) / 1–3 / ○
Russula laricina / Brown / NIS / 1–2 / ○ / ○
Basidiomycete sp.2 / Brown / NIS / ○ / 2–3 / Rhizomorph (20 to 30µm) / ○ / ○
Tuberaceae sp.1 / Brown / NIS(-IIS) / Awlc / 3–7 / ○
Cenococcum geophilum / Black / NS / 3–6 / ○ / ○

a FP, felt prosenchyma; NP, net prosenchyma; NS, net synenchyma; NIS, non-interlocking irregular synenchyma; IIS, interlocking irregular synenchyma. Terminology follows Ingleby et al. (1990).

b Older seedlings were 1 to 5 years old based on their branching pattern.

c A type of cystidia that forms bristle shapes, according to the description in Agerer (1987-2002).

The number of ECM phylotypes did not differ significantly between the seedling age groups (χ2 test, P0.05).

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Table S2. Estimated DNA sizes of the internal transcribed spacer (ITS) region and their restriction fragments from ectomycorrhizal (ECM) roots on Pinus thunbergii seedlings in a Japanese coastal forest and from ECM fruiting bodies.

Phylotype or
ECM species / ITS (bp) / Alu I restriction fragment length (bp) / Hinf I restriction fragment length (bp)
Basidiomycete sp.1 / 631 / 646 / 240/ 196
Lactarius quieticolor / 749 / 536/ 227 / 387/ 212
Clavulinaceae sp.1 / 692 / 309/ 204 / 185/ 165
Thelephoraceae sp.1 / 712 / 302 / 369
Russula sp. 1 / 717 / 507/ 223 / 377
Trichophaea sp.1 / 632 / 646 / 344/ 253
Sebacinaceae sp.1 / 643 / 616 / 359
Bankeraceae sp.1 / 589 / 321/ 151/ 92 / 355/ 286
Tomentellaceae sp.1 / 716 / 299/ 192/ 159 / 295/ 201/ 147
Russula laricina / 722 / 355/ 153 / 367/ 153
Basidiomycete sp.2 / 723 / 620/ 85 / 381
Tuberaceae sp.1 / 607 / 491 / 228/ 188/ 151
Cenococcum geophilum / 1070 / 692/ 385 / 613/ 166/ 119/ 100
Amanita pantherina a / 693 / 258/ 193/ 144/ 104 / 378
Pisolithus tinctorius a / 711 / 195 / 242/ 198
Russula emetica a / 709 / 359/ 223/ 154 / 373
Suillus granulatus a / 696 / 637/ 109 / 233/ 192/ 130

a ECM species that originated from fruiting bodies

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1c1 / 1o1 / 1o2 / 2c1 / 2c2 / 2c3 / 2o1 / 3c1 / 3o1 / 3o2 / 4c1 / 4o1 / 5o1 / 6c1 / 7o1 / 7o2 / 8c1 / 9c1 / 9c2 / 9c3 / 10c1 / 11c1 / 11c2 / 11o1 / 11o2 / 12c1 / 12o1 / 12o2 / 12o3
1c1 / 0.72 / 0.86 / 0.69 / 0.68 / 0.29 / 0.10 / 0.31 / 0.27 / 0.37 / 0.86 / 0.69 / 0.33 / 0.56 / 0.04 / 0.43 / 0.78 / 0.45 / 0.97 / 0.87 / 0.90 / 0.89 / 0.98 / 0.65 / 0.55 / 0.99 / 0.73 / 0.62 / 0.76
1o1 / 0.67 / 0.73 / 0.56 / 0.55 / 0.41 / 0.07 / 0.24 / 0.22 / 0.46 / 0.88 / 0.65 / 0.27 / 0.45 / 0.17 / 0.53 / 0.64 / 0.40 / 0.73 / 0.72 / 0.71 / 0.70 / 0.73 / 0.58 / 0.47 / 0.72 / 0.62 / 0.55 / 0.66
1o2 / 0.33 / 0.25 / 0.73 / 0.72 / 0.36 / 0.18 / 0.38 / 0.42 / 0.46 / 0.94 / 0.86 / 0.32 / 0.62 / 0.13 / 0.53 / 0.93 / 0.61 / 0.87 / 0.88 / 0.88 / 0.83 / 0.87 / 0.87 / 0.53 / 0.90 / 0.73 / 0.69 / 0.81
2c1 / 0.50 / 0.33 / 0.67 / 1.00 / 0.71 / 0.74 / 0.88 / 0.86 / 0.79 / 0.71 / 0.69 / 0.25 / 0.98 / 0.69 / 0.90 / 0.59 / 0.36 / 0.69 / 0.73 / 0.93 / 0.66 / 0.70 / 0.52 / 0.41 / 0.69 / 0.66 / 0.82 / 0.88
2c2 / 0.50 / 0.33 / 0.67 / 1 / 0.72 / 0.75 / 0.89 / 0.87 / 0.80 / 0.70 / 0.68 / 0.25 / 0.99 / 0.70 / 0.90 / 0.58 / 0.35 / 0.68 / 0.72 / 0.92 / 0.65 / 0.68 / 0.51 / 0.41 / 0.68 / 0.65 / 0.82 / 0.88
2c3 / 0.67 / 0.75 / 0.50 / 0.67 / 0.67 / 0.66 / 0.74 / 0.75 / 0.98 / 0.33 / 0.43 / 0.11 / 0.75 / 0.67 / 0.88 / 0.25 / 0.16 / 0.28 / 0.35 / 0.54 / 0.27 / 0.28 / 0.23 / 0.21 / 0.28 / 0.34 / 0.57 / 0.57
2o1 / 0.50 / 0.33 / 0.67 / 1 / 1 / 0.67 / 0.96 / 0.93 / 0.69 / 0.14 / 0.24 / 0.03 / 0.84 / 0.95 / 0.77 / 0.08 / 0.04 / 0.09 / 0.17 / 0.46 / 0.09 / 0.10 / 0.06 / 0.05 / 0.10 / 0.20 / 0.50 / 0.46
3c1 / 0.50 / 0.33 / 0.67 / 1 / 1 / 0.67 / 1 / 0.98 / 0.79 / 0.35 / 0.41 / 0.11 / 0.95 / 0.92 / 0.88 / 0.26 / 0.15 / 0.31 / 0.37 / 0.65 / 0.29 / 0.31 / 0.22 / 0.18 / 0.31 / 0.37 / 0.65 / 0.64
3o1 / 0.40 / 0.29 / 0.86 / 0.80 / 0.80 / 0.57 / 0.80 / 0.80 / 0.80 / 0.37 / 0.47 / 0.10 / 0.93 / 0.91 / 0.89 / 0.34 / 0.14 / 0.28 / 0.35 / 0.62 / 0.26 / 0.28 / 0.31 / 0.16 / 0.29 / 0.35 / 0.64 / 0.62
3o2 / 0.67 / 0.75 / 0.50 / 0.67 / 0.67 / 1 / 0.67 / 0.67 / 0.57 / 0.43 / 0.50 / 0.14 / 0.82 / 0.69 / 0.94 / 0.33 / 0.21 / 0.38 / 0.44 / 0.64 / 0.36 / 0.37 / 0.31 / 0.25 / 0.37 / 0.42 / 0.65 / 0.66
4c1 / 0.33 / 0.50 / 0.75 / 0.67 / 0.67 / 0.50 / 0.67 / 0.67 / 0.86 / 0.50 / 0.83 / 0.32 / 0.59 / 0.17 / 0.54 / 0.90 / 0.46 / 0.87 / 0.85 / 0.87 / 0.83 / 0.87 / 0.82 / 0.53 / 0.90 / 0.73 / 0.67 / 0.79
4o1 / 0.57 / 0.44 / 0.67 / 0.57 / 0.57 / 0.67 / 0.57 / 0.57 / 0.75 / 0.67 / 0.67 / 0.27 / 0.60 / 0.21 / 0.55 / 0.84 / 0.40 / 0.71 / 0.73 / 0.77 / 0.69 / 0.71 / 0.82 / 0.46 / 0.73 / 0.64 / 0.65 / 0.74
5o1 / 0.40 / 0.29 / 0.29 / 0.40 / 0.40 / 0.29 / 0.40 / 0.40 / 0.33 / 0.29 / 0.29 / 0.25 / 0.20 / 0.01 / 0.17 / 0.29 / 0.25 / 0.33 / 0.32 / 0.32 / 0.32 / 0.34 / 0.25 / 0.20 / 0.33 / 0.27 / 0.24 / 0.29
6c1 / 0.50 / 0.33 / 0.67 / 1 / 1 / 0.67 / 1 / 1 / 0.80 / 0.67 / 0.67 / 0.57 / 0.40 / 0.80 / 0.92 / 0.48 / 0.29 / 0.56 / 0.61 / 0.85 / 0.53 / 0.56 / 0.42 / 0.33 / 0.56 / 0.56 / 0.78 / 0.82
7o1 / 0.40 / 0.57 / 0.57 / 0.80 / 0.80 / 0.57 / 0.80 / 0.80 / 0.67 / 0.57 / 0.86 / 0.50 / 0.33 / 0.80 / 0.81 / 0.03 / 0.02 / 0.04 / 0.12 / 0.40 / 0.04 / 0.04 / 0.03 / 0.02 / 0.04 / 0.16 / 0.47 / 0.42
7o2 / 0.33 / 0.75 / 0.50 / 0.67 / 0.67 / 0.75 / 0.67 / 0.67 / 0.57 / 0.75 / 0.75 / 0.44 / 0.29 / 0.67 / 0.86 / 0.39 / 0.24 / 0.44 / 0.51 / 0.73 / 0.43 / 0.44 / 0.36 / 0.27 / 0.44 / 0.49 / 0.73 / 0.74
8c1 / 0.50 / 0.33 / 0.67 / 0.50 / 0.50 / 0.33 / 0.50 / 0.50 / 0.80 / 0.33 / 0.67 / 0.57 / 0.40 / 0.50 / 0.40 / 0.33 / 0.40 / 0.79 / 0.75 / 0.75 / 0.75 / 0.79 / 0.91 / 0.47 / 0.85 / 0.63 / 0.56 / 0.67
9c1 / 0.40 / 0.29 / 0.57 / 0.40 / 0.40 / 0.29 / 0.40 / 0.40 / 0.33 / 0.29 / 0.29 / 0.25 / 0.67 / 0.40 / 0.33 / 0.29 / 0.40 / 0.46 / 0.53 / 0.45 / 0.45 / 0.46 / 0.43 / 0.29 / 0.46 / 0.39 / 0.35 / 0.42
9c2 / 0.50 / 0.33 / 0.33 / 0.50 / 0.50 / 0.33 / 0.50 / 0.50 / 0.40 / 0.33 / 0.33 / 0.29 / 0.40 / 0.50 / 0.40 / 0.33 / 0.50 / 0.40 / 0.95 / 0.90 / 0.89 / 1.00 / 0.73 / 0.71 / 0.97 / 0.74 / 0.64 / 0.78
9c3 / 0.33 / 0.25 / 0.75 / 0.67 / 0.67 / 0.50 / 0.67 / 0.67 / 0.57 / 0.50 / 0.50 / 0.44 / 0.29 / 0.67 / 0.57 / 0.50 / 0.33 / 0.57 / 0.67 / 0.88 / 0.84 / 0.94 / 0.80 / 0.83 / 0.88 / 0.73 / 0.68 / 0.80
10c1 / 0.50 / 0.33 / 0.67 / 1 / 1 / 0.67 / 1 / 1 / 0.80 / 0.67 / 0.67 / 0.57 / 0.40 / 1 / 0.80 / 0.67 / 0.50 / 0.40 / 0.50 / 0.67 / 0.85 / 0.90 / 0.65 / 0.53 / 0.91 / 0.76 / 0.80 / 0.91
11c1 / 0.50 / 0.33 / 0.33 / 0.50 / 0.50 / 0.33 / 0.50 / 0.50 / 0.40 / 0.33 / 0.33 / 0.29 / 0.40 / 0.50 / 0.40 / 0.33 / 0.50 / 0.40 / 0.50 / 0.33 / 0.50 / 0.89 / 0.65 / 0.52 / 0.89 / 0.95 / 0.82 / 0.75
11c2 / 0.50 / 0.33 / 0.33 / 0.50 / 0.50 / 0.33 / 0.50 / 0.50 / 0.40 / 0.33 / 0.33 / 0.29 / 0.40 / 0.50 / 0.40 / 0.33 / 0.50 / 0.40 / 1 / 0.67 / 0.50 / 0.50 / 0.72 / 0.69 / 0.98 / 0.74 / 0.64 / 0.78
11o1 / 0.33 / 0.25 / 0.75 / 0.33 / 0.33 / 0.25 / 0.33 / 0.33 / 0.57 / 0.25 / 0.50 / 0.44 / 0.29 / 0.33 / 0.29 / 0.25 / 0.67 / 0.57 / 0.67 / 0.75 / 0.33 / 0.33 / 0.67 / 0.68 / 0.71 / 0.57 / 0.51 / 0.61
11o2 / 0.80 / 0.57 / 0.29 / 0.40 / 0.40 / 0.57 / 0.40 / 0.40 / 0.33 / 0.57 / 0.29 / 0.50 / 0.33 / 0.40 / 0.33 / 0.29 / 0.40 / 0.33 / 0.8 / 0.57 / 0.40 / 0.40 / 0.80 / 0.57 / 0.55 / 0.45 / 0.39 / 0.48
12c1 / 0.50 / 0.33 / 0.67 / 0.50 / 0.50 / 0.33 / 0.50 / 0.50 / 0.80 / 0.33 / 0.67 / 0.57 / 0.40 / 0.50 / 0.40 / 0.33 / 1 / 0.4 / 0.5 / 0.33 / 0.50 / 0.50 / 0.50 / 0.67 / 0.40 / 0.73 / 0.63 / 0.77
12o1 / 0.40 / 0.29 / 0.57 / 0.80 / 0.80 / 0.57 / 0.80 / 0.80 / 0.67 / 0.57 / 0.57 / 0.50 / 0.33 / 0.80 / 0.67 / 0.57 / 0.40 / 0.33 / 0.40 / 0.57 / 0.80 / 0.80 / 0.40 / 0.29 / 0.33 / 0.40 / 0.93 / 0.71
12o2 / 0.40 / 0.29 / 0.57 / 0.80 / 0.80 / 0.57 / 0.80 / 0.80 / 0.67 / 0.57 / 0.57 / 0.50 / 0.33 / 0.80 / 0.67 / 0.57 / 0.40 / 0.33 / 0.40 / 0.57 / 0.80 / 0.80 / 0.40 / 0.29 / 0.33 / 0.40 / 1 / 0.78
1203 / 0.40 / 0.29 / 0.57 / 0.80 / 0.80 / 0.57 / 0.80 / 0.80 / 0.67 / 0.57 / 0.57 / 0.50 / 0.33 / 0.80 / 0.67 / 0.57 / 0.40 / 0.33 / 0.40 / 0.57 / 0.80 / 0.40 / 0.40 / 0.29 / 0.33 / 0.40 / 0.67 / 0.67

Table S3. Morisita-Horn index (upper half of the matrix) and Sørensen index (lower half of the matrix) for the similarity among pairs of pine seedlings collected from a Japanese coastal pine forest.

Index values for the similarity between current-year and older seedlings within the same plot are shown in boldface. The seedling identification comprises three letters or groups of characters: the first, second, and third groups indicate the plot number, seedling age (c, current year; o, older), and sample number, respectively. Rows that contain the data for current-year seedlings are shaded.

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References

Agerer R (1987–2002) Colour Atlas of Ectomycorrhizae. Einhorn-Verlag, Schwabisch Gmund

Ingleby K, Mason PA, Last FT, Fleming LV(1990) Identification of Ectomycorrhizas. HMSO, London

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