ITS sequence was used as a barcode to identify Lasiosphaera Calvatia and its adulterants.Methods:We collected 21 samples of Lasiosphaera Calvatia and extracted their DNA，then amplified their ITS sequences.We also downloaded sequences of Lasiosphaera Calvatia’s adulterants from GenBank.Sequences were assembled and analysis their variable sites.The Kimura 2-Paramter(K2P)genetic distances and compute the Neighbor-joining(NJ)phylogenetic tree were computed.Results:The maximum intraspecific K2P genetic distances of the ITS sequence of Calvatia gigantea(Batsch ex Pers.)Lloyd is 0.The maximum intraspecific K2P genetic distances of the ITS sequence of Lasiosphaera fenzlii Reich.is 0.003.The maximum intraspecific K2P genetic distances of the ITS sequence lengths of Calvatia lilacina(Mont.et Berk.)Lloyd is 0.003.The minimum interspecific K2P genetic distances of Calvatia gigantea and adulterants is 0.019，Lasiosphaera fenzlii and adulterants is 0.031，while Calvatia lilacina and adulterants is 0.634.The maximum intraspecific genetic distances of Calvatia gigantea、Lasiophaera fenzlii and Calvatia lilacina were all less than the minimum interspecific genetic distances of themselves and their adulterants.the NJ tree based on ITS sequence showed that Calvatia gigantea、Lasiosphaera fenzlii and Calvatia lilacina can gather into a branch sperately.Conclusion:ITS sequence was able to identify Lasiosphaera Calvatia from their adulterants,which is suitable in the identification of Lasiosphaera Calvatia and its adulterants.