Abstract:Based on traditional morphological classification and ITS (Internal transcribed spacer) sequences analysis, the company classified and scribed? two wild macrofungi (sample numbers: HBES1003, HBES1096) collected from Enshi, Hubei Province, and analyzed their phylogeny. The two strains were identified as Xerocomus species based on their morphological characteristics. In order to further determine the accuracy and scientific nature of identification, PCR amplification and sequencing of ITS whole region sequences of the samples of the two strains were performed, Blast comparison was performed in NCBI database, and the phylogenetic tree was constructed by NJ(Neighbor-Joining) adjacency method. It was analyzed and identified by exploring its correlation with known strains. The results showed as follows: Sample HBES1003 was Xerocomus subtomentosus and sample HBES1096 was Xerocomus nigromaculatus.