Tricholoma matsutake, one of the most famous edible mushroom in Asia, has been cultivated in red pine forest. Because of its difficulty in artificial cultivation, T. matsutake cultivation has relied on foraging in the forest. Under certain environmental conditions, T. matsutake form the Shiro, the condensed mycelium, and develop into fruiting bodies. Among the certain environmental conditions, fungal communities play a major role in the mushroom development. Therefore, fungal community investigations for Bonghwa and Yangyang have been conducted on soil with fairy rings in the past, soil with existing fairy rings, and soil with presumptive fairy rings developing in the future. From the six soil samples, total 163 genera of fungi were detected and species diversity and species abundance of each sample were also analyzed. In result, the species diversity and species abundance of the fairy rings in Yangyang were lower than those in Bonghwa. In comparison with the fairy rings in Yangyang and Bonghwa, the dominance of the genus Tricholoma was higher and that of the genus Motilella was lower in Yangyang. Through the continuous study, establishment of optimal environmental conditions to promote the T. matsutake cultivation is highly expected.

Tricholoma matsutake is an economically important ectomycorrhizal mushroom, but artificial cultivation remains challenging. While much research has been conducted on the bacteria and fungi associated with T. matsutake, studies on archaea have been limited. This study compared and analyzed the diversity and structural differences of archaeal communities in the fairy ring and non-fairy ring soils using a metabarcoding approach. The alpha diversity analysis revealed that the archaeal community in fairy ring soil had lower diversity compared to non-fairy ring soil, and beta diversity analysis clearly separated the community structures between the two soil types. Among all analyzed soils, Nitrososphaeria (Nitrososphaerota) dominated the archaeal community at class level. In fairy ring soils, Nitrosotalea (Nitrosotaleaceae) was predominant, while in non-fairy ring soils, Nitrosocaldaceae dominated. These results suggest that Nitrosotalea may have a close association with T. matsutake, highlighting the need for further in-depth research on archaea as potential growth promoting microorganism for T. matsutake mycelial growth.

As a member of ectomycorrhizal fungi, Tricholoma matsutake has a symbiotic relationship with its host, Pinus densiflora. To cultivate T. matsutake artificially, the co-cultivation of T. matsutake mycelia and bacteria from shiro was introduced. In this study, bacteria were isolated from soil samples in Bonghwa-gun, and seven bacterial isolates (B22_7_B05, B22_7_B06, B22_7_B07, B22_7_B08, B22_7_B10, B22_7_B13, and B22_7_B14) promoted the growth of T. matsutake mycelia (147.48, 232.11, 266.72, 211.43, 175.17, 154.62, and 177.92%, respectively). Sequencing of the 16S rRNA region of the isolated bacteria was performed. B22_7_B05 and B22_7_B10 were identified as Bacillus toyonensis, B22_7_B06 and B22_7_B08 as Paenibacillus taichungensis, B22_7_B07 and B22_7_B14 as P. gorilla, and B22_7_B13 as P. odorifer. These bacterial isolates were associated with the shiro community and are expected to contribute to the cultivation of T. matsutake.

Tricholoma matsutake is a traditional favorite food in East Asia, cultivated in fairy rings called “shiro,” which are found near Pinus densiflora. For effective artificial cultivation of Tri. matsutake, microorganisms from symbiotic fairy rings are co-cultivated. In this study, one bacterial isolate (Y22_B35) and two fungal isolates (Y22_F64 and Y22_F68) displayed growth-promoting effects on Tri. matsutake mycelium (158.47, 125.00, and 122.26% enhanced growth, respectively). For identification, 16S rRNA or ITS regions from the microorganisms¡¯ genomes were sequenced. Other sequences, including BenA, CaM, and RPB2 were sequenced in the fungal isolates. The bacterial isolate Y22_B35 was identified as Bacillus cereus. Y22_F64 and Y22_F68 were identified as Umbelopsis nana and Aspergillus parvulus, respectively. To identify the effects of the dominant microorganisms on Tri. Matsutake cultivation, metagenomic analyses were performed. Discovery of these Tri. matsutake mycelium growth-promoting microorganisms and metagenomics analyses are expected to contribute to our understanding of Tri. matsutake fruiting body growth and construction of biomimicry.