Abstract: DNA was extracted from the fruiting bodies of 24 wild Lactarius sensu lato collected from different locations in Enshi area. The sequencing results were compared with GenBank database, and the comparison results were drawn into a phylogenetic tree for analysis. The results showed that the similarity between the tested strains and the nearest homologues reached 99%~100%. The genetic distance between the target strain and the nearest homolog was analyzed. It was found that the genetic distance between S1, S2, S3, S4, S5, S7, S8, S9, S10, S11, S12, S13, S14, S16 and S22 and the three nearest Lactarius vividus was 0, while S15 was 0.001 and S23 was 0.003. The genetic distance between S6, S17, S21 and Lactarius salmonicolor （JX852629.1） was 0. The genetic distance between S19, S20 and three different species of Lactarius hatsudake was 0, while there was a certain genetic distance between S18 and Lactarius hatsudake. The genetic distance between S24 and Lactifluus pilosus （MZ157882.1） was 0. The common wild mushrooms in Enshi were Lactarius vividus, Lactarius salmonicolor and Lactarius hatsudake, which were edible wild mushrooms. There was also a confusing poisonous fungus-Lactifluus pilosus.

Key words: Lactarius sensu lato, ITS sequencing, systematic development, genetic distance