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.

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.

The beneficial effects of EM on plant growth, yield, quality in agriculture have been consistently demonstrated. In this study, EM of four types used is Lactobaillus sp., saccharomyces cerevisiea, Rhodobacter sp. and Bacillus sp.. Bacillus sp. inhibited Mycelium growth of the oyster mushroom in agar plate. Bacillus sp. is known to suppress plant pathogenic microorganism in soil. The other three type of EM did not inhibit mycelium growth of oyster mushroom. Also when filtering media of Lactobacillus sp. and saccharomyces cerevisiea cultivation add to liquid media(PDB) of oyster mushroom, mycelium growth was a little faster than control. In the column test, Mycelium growth of the medium inoculated with 0.5% saccharomyces cerevisiea was faster than the control. In the substrate inoculated Rhodobacter sp. was similar to the control. In the substrate inoculated Lactobacillus sp. of higher concentration, mycelium growth of the oyster mushroom was showed late than control.

본 연구에서는 효율적으로 팽나무버섯 액체종균을 생산할 때에 폭기(공기방울을 매우 작게하여) 배양의 효과에 대하여 액체 배지 내 이산화탄소 농도와 잔류 유리당 함량 변화로써 측정하였다. 이산화탄소 농도는 폭기 진행 중 배양용기의 배출구에서 측정하였다. 액체종균은 폭기 배양 3일째에 균사생장이 급격히 증가하였으며, 이산화탄소 농도(CO2)도 급증하였으나 폭기 5일째에는 정점을 나타낸 후 감소하였다. 액체배지 내 환원당은 배양 7일째까지 일정 속도로 감소되었고 이후에는 검출되지 않았고, 침전 균사체의 중량은 3일째 이후 비슷한 수준으로 유지되었다. 총질소(T-N)량은 폭기 11일째까지 일정한 수준으로 감소되었다. 유리당의 함량은 폭기 배양 7일째에 가장 낮았으므로 폭기배양 6~7일째에 접종원으로 사용하는 것이 가장 효과적인 것으로 생각되었다. 그리고 액체종균의 배양 중에 배출구에서의 이산화탄소 농도 측정은 저비용으로 액체종균의 균사배양 정도를 추정하는 간이 지표로 이용할 수 있을 것으로 판단되었다.

Mushroom is cultivated as one of the major economical crops in many areas of the Korea. The total production have steadily increased approximately 151,913 M/T in 2000 to 186,400 M/T in 2007. This study was carried out to investigate applicability of mushroom production using various organic media resources within the country. Eight organic resources were collected from various areas. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at the media of 10% red ginseng marc, 20% lacquer tree, 20% Juglans mandshurica, 10% Cudrania tricuspidata, 10~20% Acer pensylvanicum, 10% Lindera glauca. Mushroom mycelial growth at red ginseng marc media was slower than that of the control. But the sponin of the red ginseng was not detected at the fruiting body grown from red ginseng marc media, And three organic resources(barly powder, sweet potato powder, potato powder) was used to substitute rice bran used in mushroom cultivation. Pleurotus ostreatus and Fulammulina velutipes showed the highest growth at 10~30% sweet potato powder, 20% barly powder and 10% potato powder.

Ionizing treatments were applied at 5Gy, 10Gy, 50Gy, 100Gy and 500Gy to mushroom mycelium (Lentinula edodes) in order to assesss the effect of the gamma-ray radiation. We mutated by gamma irradiation, 12 strains were isolated. 9 strains of 12 strains were appeared antagonistic interaction on solid medium. Growth rate of mycelium in JMIR-3 and JMIR-6 strains were similar to their control. But, the growth rate of strain JMIR-1, JMIR-2, JMIR-4, JMIR-5, JMIR-7, JMIR-8, JMIR-9, JMIR-10, JMIR-11 and JMIR-12 had generally different compared to control. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 696 to 780bp. The reciprocal homologies of the ITS region sequences in 5Gy irradiation group (JMIR-5, JMIR-6, JMIR-7 and JMIR-8) and 10Gy irradiation group (JMIR-9, JMIR-10, JMIR-1 and JMIR-12) indicated 99%. The reciprocal homologies of the ITS region sequences in 500Gy irradiation group (JMIR-1, JMIR-2, JMIR-3 and JMIR-4) indicated from 95 to 99%. It seemed that mycelium growth and ITS sequence could be changed the irradiation dose of the gamma-ray radiation.

Effect of trehalose for the mycelial growth of Tricholoma matsutake were examined. When T. matsutake Z-1 strain was cultured in the partially modified matsutake liquid (PMML) medium and the Hamada matsutake liquid (HML) medium supplemented with trehalose at 24 ℃ for 80days, the vegetative mycelial dry weights showed higher value compared with those of PMML medium and HML medium supplemented with glucose (control). The range of the effect of 1.0-8.0% carbohydrate substrate on vegetative mycelial growth was investigated. The optimal concentration for mycelial growth was 2.0% for the glucose medium but 8.0% for the trehalose medium. To evaluate the potential of the production of trehalase from T. matsutake, the extracellular trehalase activity during the vegetative mycelial growth was measured. The activity of the extracellular trehalase increased during vegetative mycelial growth of T. matsutake and was maximal 70 days after inoculation. This extracellular enzyme was investigated for the purification and the characterization. The partially purified trehalase was obtained from about 1.53l static culture filtrate, with 19.1% recovery, and about 2,940 fold purification. The molecular mass was about 62.6kDa (SDS-PAGE) and 70kDa (Gel-filtration). The enzyme was most active around 40℃ and pH 5.0 and stable over a pH of 4.5~ 6.5 for 30min at 37℃. The enzyme readily hydrolyzed trehalose having α -1,1 glucosidic bond. However, it did not hydrolyze disaccharides such as maltose, iso-maltose, cellobiose, saccharose and lactose.