The study of the phylogeny of species is a basic discipline for the effective protection, development, and utilization of their germplasm resources. Since this century, the fungi of the Agrocybe genus have become the most successful class of medium and high-grade edible and medicinal fungi that have been artificially domesticated and grown on a large scale, and have high commercial value. However, there are few reports on its germplasm resources. To clarify the genetic and developmental relationship at the genus level of the Agrocybe in the world. In this paper, Based on the NCBI Nucleotide database(National Center for Biotechnology Information), taking the single gene ITS sequence as the research object, the T92 (Tamura 3-parameters) parameter model was used to calculate the genetic distance, and the Neighbor-Joining Method was used to construct the genus-level phylogenetic tree of the Agrocybe. The results showed that the phylogenetic tree constructed by 192 ITS single gene sequences of 43 species of the Agrocybe (including 31 known species and 12 undetermined species) in the NCBI Nucleotide database was roughly divided into 6 branches and belonged to different groups, and 17 undetermined species of the Agrocybe sp. was preliminarily identified. This study has a certain reference value for the genetic relationship among different species of Agrocybe all over the world, as well as the selection of the parent bank and the protection of germplasm resources of each species of the genus.