Abstract: A pair of general primer (ITS4ITS5) and a pair of specific primer (TMFTMR) were used to test in the amplification of the internal transcribed spacer (ITS) region of the chromosomal DNA of fruit bodies of Tricholoma matsutake and its 6 isolates,Tricholoma bakamatsutake and its 3 isolates,fruit bodies of Flammulina velutipes,Pleurotus ostreatus and Agaricus bisporus.The primers of ITS4 and ITS5 amplified about 600?bp fragments for all samples.However,with TMF and TMR primers we can amplify about 500?bp fragments only from the fruit bodies of Tricholoma matsutake and their corresponding isolates.The ITS fragments of Tricholoma matsutake fruit body (TF25) and its corresponding isolate (TM251122) were sequenced and aligned.Their sequences showed a 100% homogeneity.Based on these facts,6 isolates designated as TM06112,TM151101,TM251122,TM281121,TM291121 and TM30122 were confirmed to be pure cultures of Tricholoma matsutake.

Key words: Tricholoma matsutake