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.

가송이(Tricholoma bakamatsutake Hongo)는 주름버섯목(Agaricales), 송이과(Tricholomataceae)에 속 하는 외생균근성 버섯류의 하나로, 송이(T. matsutake)와 일반적인 외형이 거의 비슷하며, 송이향과 맛이 강하게 나기 때문에 이 두 균종은 쉽게 혼동되며, 실제 분류 및 계통발생학적으로도 가송이와 송이는 유연 관계가 있는 것으로 밝혀졌다. 가송이는 한국, 일본, 대만, 중국의 신갈나무 등과 같은 활엽수림에 분포하는 것으로 알려져 있으며, 최근에는 제주도 구실잣밤나무림에서 발견되었다. 가송이는 균사생장이 매우 느려 연구에 어려움이 많아 균사배양 최적 조건을 구명하고자 본 연구를 실시하였다. 온도에 따른 가송이 균주 별 균사생장 특성을 조사한 결과, 모든 균주에서 25℃에서 가장 균사생장속도가 가장 빨랐으며, 특히 3833 균주가 다른 균주에 비해 약 1.5배 빠른 것으로 조사되었다.

To cultivate pine mushroom (Tricholoma matsutake) artificially, co-cultivation with microorganisms has been introduced. Here, experiments were performed to assess the growth-promoting effect of bacteria on T. matsutake mycelia. Bacteria were isolated from soil samples collected in Yangyang County, Korea. Four of the bacterial isolates (Y22_B06, Y22_B11, Y22_B18, and Y22_B22) exhibited a growth-promoting effect on T. matsutake mycelia (154.67%, 125.91%, 134.06%, and 158.28%, respectively). To analyze the characteristics of the bacteria, especially the antifungal activity, -amylase and cellulase activity assays were performed. In comparison with the controls, the isolated bacteria exhibited low -amylase and cellulase activity. 16S rRNA gene sequencing was performed to identify the four bacterial isolates. The isolates belonged to the Terrabacteria group and were identified as Microbacterium paraoxydans, Paenibacillus castaneae, Peribacillus frigoritolerans, and P. butanolivorans. These bacterial isolates are expected to have contributed to the growth promotion of T. matsutake mycelia and the artificial cultivation of T. matsutake.

This study was conducted to develop a renewable and sustainable bio-material to replace polystyrene (EPS) in fungal-mycelium-based composite using agricultural by-products. Four mushrooms (Ganoderma lucidum, Fomitella fraxinea, Phellinus linteus, and Schizophyllum commune) were cultured in an oak sawdust plus rice bran substrate to select the mushroom with the best growth. The mycelia of G. lucidum showed the best growth. To investigate the optimal mixing ratio with spent mushroom substrate (SM) and oak sawdust (OS), samples were prepared by mixing SM and OS at ratios of 50%:50%, 60%:40%, and 80%:20% (w/w). Each substrate was then inoculated with G. lucidum. G. lucidum showed the best mycelial growth of 140.0 mm in the substrate with SM and OS mixed at a 60%:40% ratio. It was also found that the substrate with SM and OS mixed at a 60%:40% ratio had the best handling properties. The compressive strength of mycelial materials inoculated with G. lucidum was in the range of 300–302 kgf mm-1, and the materials were four times stronger than polystyrene materials. These results indicate that substrates comprising spent mushroom substrate mixed with oak sawdust can be successfully upcycled to mycelium-based composite materials using G. lucidum. This represents a sustainable approach.

The importance of biocomposites has increased owing to the changes in global consumption trends and rapid climate change. Technologies using mushroom mycelium cultivation, and molding methods for mycelial application have gained attention as potential strategies for producing eco-friendly composites. Currently, mushroom mycelia are used as raw materials for food and cosmetics; however, research on their utilization as biocomposite materials is limited. Therefore, the potential for the development of mushroom mycelium-related products and technologies is high. This review analyzes the domestic and international patent application trends related to the technologies for composite (packaging, insulation, adhesives, and leather) and food (substitute for meat) materials using mushroom mycelium, as an eco-friendly biocomposite material, to provide objective patent information that can further research and development (R&D) in this field.

In this study, we aimed to compare the mycelial growth of Pleurotus ostreatus after medium supplementation with various amino acids at different concentrations to select the optimal medium nutrient composition for mycelial growth. The mycelial growth of P. ostreatus was investigated after adding four amino acids (tryptophan, threonine, methionine, and lysine) at 0.5% or 1% to the medium.The rate of P. ostreatus mycelial growthwas faster in the potato dextrose agar (PDA) medium supplemented with threonine at 0.5% or 1% than that of the control, whereas it was inhibited by tryptophan treatment. Supplementation of sawdust mediumwith all amino acids, except tryptophan, at 0.5% did not alter the mycelial growth, compared to the controls. However, addition of any amino acid to sawdust medium at a higher concentration (1%) inhibited the mycelial growth. The laccase acitivity of P. ostreatus mycelium cultured in PDA medium was the highest when threonine was added, and the lowest when tryptophane was added, consistent with the results of the mycelial growth. Therefore, the addition of threonine, methionine, or lysine to PDA medium at a concentration of 0.5-1%was effective for increasing the mycelial growth of P. ostreatus; however, it inhibited mycelial growth insawdust medium, suggesting that the effects of amino acids dependedon the medium nutrient composition.

To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.

After liquid culture of Phellinus baumii (P. baumii) mycelium (LPBM) was prepared, LPBM was fractionated into A∼E fraction (A; hot-water extract of liquid culture including mycelia, B; crude polysaccharide of A, C; hot-water extract of mycelia, D; crude polysaccharide of C, and E; crude polysaccharide of culture broth) to evaluate for possibility as functional materials with immunostimulatory activity. In macrophage stimulatory activity, E fraction as postbiotics significantly increased secretion of NO and IL-12 from RAW 264.7 cells. Next, when the splenocytes of C3H/HeN mice were primary cultured, E fraction showed significantly mitogenic activity with enhancing mitogen-related cytokines (IFN-γ and TNF-α) production from splenocyte. E fraction also potently stimulated GM-CSF production from Peyer’s patch cells as well as Peyer’s patch-mediated bone marrow cell proliferation. In addition, the immunostimularoy E fraction contained neutral sugar (73.8%), uronic acid (10.6%), protein (7.8%), and polyphenol (7.5%), and mainly consisted of glucose (39.1%), galactose (21.7%), mannose (11.1%), galacturonic acid (9.9%), and arabinose (8.9%) as component sugars. In conclusion, it was demonstrated that postbiotics including exopolysaccharide fractionated from liquid culture of the P. baumii mycelium could enhanced immunostimulatory activity.