The main objectives of this research were to study the cultural and nutritional characteristics of Tricholoma matsutake and to establish its liquid culture system. The optimum growth of T. matsutake was observed in HA and TMM agar media. Similarly highest growth was observed in PDB and TMM liquid media. The optimal temperature for the mycelial growth was 25 oC. The most suitable carbon source was dextrin among 12 different carbon sources tested. Yeast extract and peptone were best nitrogen sources among 17 different sources tested. The optimum mineral salts were Fe₂(SO₄)₃·H₂O and KCl among 9 different sources tested. Shaking culture gave higher mycelial growth compared to stationary culture. Similarly, optimum medium amount for shaking culture was 100 ml per 250 ml flask. The highest mycelial growth was obtained when 5~7 mycelial discs were inoculated in 100 ml of medium and incubated for 8~9 weeks, respectively. The highest proportion of mycelial growth was observed at 40 : 1 ratio of medium to inoculum volume in 8 l air-lift fermenter.