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<i>Asproinocybe</i> and <i>Tricholosporum</i> are not well known, and their placement at the family level remains undetermined. In this study, we conducted molecular phylogenetic analyses based on nuc rDNA internal transcribed spacer region (ITS) and nuc 28S rDNA (nrLSU), and a dataset with six molecular markers (ITS, LSU, RNA polymerase II largest subunit (RPB1), RNA polymerase II second largest subunit (RPB2), 18S nuclear small subunit ribosomal DNA (nrSSU), and translation elongation factor 1-alpha (TEF1-α)) using Bayesian (BA) and Maximum Likelihood (ML) methods, we found that the species of <i>Asproinocybe</i> and <i>Tricholosporum</i> formed an independent family-level clade (0.98/72). Asproinocybaceae fam. nov., a new family, is established here for accommodating this clade. Two new species, <i>Asproinocybe sinensis</i> and <i>Tricholosporum guangxiense</i>, from subtropical and tropical karst areas of China, are also described here.