Matsutake mushroom Mycorrhizal isolationNuclear ribosomal RNA gene (rDNA) IGS1 region a b s t r a c t Tricholoma matsutake is one of the most highly valued ectomycorrhizal mushrooms, primarily associated with conifers. Here, we examined the association of T. matsutake with hemlock and fir species native to the subalpine forests of Japan. Basidiomata of T. matsutake were harvested from the forests of Honshu Island, Japan, along with two soil samples directly beneath the basidiomata; ectomycorrhizal root tips were also collected, and used for fungal isolation and molecular analyses. Mycorrhizal fungi were isolated from the roots of Abies veitchii (Veitch's fir) and Tsuga diversifolia (northern Japanese hemlock). These fungi were identified as T. matsutake based upon their production of whitish mycelial colonies on nutrient agar media, and confirmed using molecular analyses; the ectomycorrhizal association between T. matsutake and A. veitchii was also confirmed by mycorrhizal synthesis in vitro. This work represents the first description of T. matsutake ectomycorrhizations withAbies and Tsuga species, as well as a detailed morphological description of these associations. /locate /m yc m y c o s c i e n c e 5 6 ( 2 0 1 5 ) 4 0 2 e4 1 2 http://dx.

Matsutake (Tricholoma matsutake) is a commercially valuable edible ectomycorrhizal mushroom. The physiological traits of T. matsutake have been previously assessed using mycelial isolates isolated from basidiomata; however, few studies have focused on basidiospores. Here, we report that sibling T. matsutake isolates generated from basidiospores on a single basidioma show distinct physiological variation. We first established 145 isolates of T. matsutake on modified Norkrans' C (MNC) agar medium and found that their radial growth varied significantly. The mycelial biomasses of nine isolates with different growth rates were reduced on low-carbon and low-nitrogen MNC media. However, the colony diam of one isolate was significantly elevated on low-carbon medium, and the colony diam of two isolates were significantly elevated on low-nitrogen medium. In co-cultures of two or three isolates, commensal and amensal interactions were observed. The physiological variation induced by low carbon and nitrogen levels and the mycelial interactions between sibling isolates imply mechanisms for the genetic and functional characteristics of mycelia of T. matsutake.

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