Patent ID: 7269923

Claim:
A method of preparing a Tricholoma matsutake -infected young pine tree, comprising the steps of: inoculating fungal mycelia obtained by pulverizing Tricholoma matsutake fruit bodies liquid-cultured in potato dextrose broth medium into the bottom of a sterilized culture container at an amount of 0.01-0.02 mg dry weight/mL sterile water; mixing perlite and sphagnum peatmoss at a ratio of 80:1-2, and placing the resulting mixed soil onto the inoculated fungal mycelia; preparing K-liquid medium containing 1.65 g of NH 4 NO 3 , 0.2 g of KNO 3 , 0.002 g of CaCl 2 .2H 2 O, 0.02 g of KCI, 0.2 g of KH 2 PO 4 , 0.9 g of MgSO 4 .7H 2 O, 0.2 g of (NH 4 ) 2 HPO 4 , 0.5 g of NH 4 -Tar, 0.5 ml of Fe-Cit, 0.031 g of H 3 BO 3 , 0.01516 g of MnSO 3 .4H 2 O, 0.0086 g of ZnSO 4 .7H 2 O, 0.00083 g of KI, 0.00025 g of Na 2 MoO 4 .2H 2 O, 100 μg of Thiamine HCl, 1.0 g of malt extract, 0.5 g of yeast extract, 0.3 g of casein and 3.0 g of glucose per 1 L in water, adjusting pH of the medium to pH 5.5-5.6, and aliquotting the K-liquid medium onto the mixed soil; aseptically germinating pine seeds up to 3 cm in length, planting the resulting aseptic seedlings into infection medium containing the mixed soil and the K-liquid medium, and covering the culture container with a lid; and coculturing the pine seedling and the Tricholoma matsutake mycelia at 15-25° C. for 24 hrs under 10-40,000 lux light intensity.