Abstract: Abrsat: This study focused on 14 edible fungi species—including Flammulina velutipes, Volvariella volvacea, Pleurotus ostreatus, Lyophyllum decastes, Agrocybe cylindracea, Lentinula edodes, Phallus indusiatus, etc. The ash content, protein content, crude polysaccharide content, amino acid composition and content of these edible fungi were systematically determined and analyzed. The volatile flavor substances were analyzed using HS-SPME/GC-MS, while the non-volatile flavor components such as free amino acids and 5'-nucleotides were also evaluated. The results showed that among the 14 edible fungi, the protein content of Agrocybe cylindracea was the highest, reaching 40.12 g/100 g, the ash content of Volvariella volvacea was significantly higher than that of other species. Tricholoma matsutake and Agaricus subrufescens contained the highest polysaccharide content, reaching 11.77 g/100 g. More than 150 volatile substances were detected, with 60 kinds of volatile components being the richest in Tricholoma matsutake, and the key flavor substances of 14 edible fungi were mainly alcohols and aldehydes, including 1-octen-3-ol and Isovaleraldehyde. The results of the determination of non-volatile substances showed that the total content of flavor nucleotides ranged widely, with the highest total content in Volvariella volvacea, followed by Agaricus bisporus, Tricholoma matsutake and Pleurotus ostreatus, with contents of 3.45, 2.50, 2.42 and 2.35 mg/g respectively, all categorized as medium levels. Agaricus bisporus had the highest Glu content, and its equivalent umami concentration (EUC) reached 1142.12 g MSG/100 g, belonging to the top umami grade. The main organic acid was succinic acid, which had the highest content in Volvariella volvacea, Agrocybe cylindracea and Agaricus subrufescens. Principal component analysis (PCA) and cluster analysis indicated that there were significant differences in the nutritional and flavor components of different edible fungi. Volvariella volvacea, Agrocybe cylindracea and Agaricus bisporus formed independent clusters due to their unique nutrient compositions. This study revealed the nutritional composition and flavor characteristics of different edible fungi through multi-faceted analysis, providing a theoretical basis for the development of high-value-added products and functional food design.