Abstract: Lepista sordida which was collected from Genhe District in Inner Mongolia was used as materials， tissue isolation was used to isolate the tissue of pileus， stipe， and junction of them. The optimal culture medium and the best tissue for isolation was investigated by measuring growth rate and pollution rate. Then the isolate was identified by internal transcribed spacer （ITS）， the genetic distance was calculated by Kimura2-parame-ter， and a NJ systematic tree was established by the neighbor-joining method. The results showed that the optimal culture medium was PDA added with sporocarp boiled water， and the junction of pileus and stipe showed fast growth and low pollution， which was the best isolation position. Moreover， the pollution rate could be effectively reduced when the sporocarp had been air-dried for 2 days. At last， the isolate was identified as Lepista sordida by the analysis of phylogenetic relationship of ITS gene sequences.

Key words:  Lepista sordida, tissue isolation, ITS sequence analysis, phylogenetic analysis