Agrocybe cylindracea is an important fungi mushroom, highly valued as a functional food for its nutritional and me-dicinal benefits. Many bioactive extracts from A. cylindracea have been found to exhibit antitumor and antioxidant activities. This research investigated the distinct substrates that affected the physicochemical properties and yield of A. cylindracea. The substrates used were 48% Giant Juncao Grass and 30% Dicraptoris Dicholoma grasses, 48% Wasted Tea Leaves and 30% Sawdust, and 78% Cottonseed Hulls, all supplemented with 20% Wheat Bran and 2% Lime. The findings indicated the cotton seed husk marked a significant in Agrocybe cylindricae yield, followed by the wasted tea leave+sawdust. Ash content peaked in the fruiting bodies grown on Giant Juncao Grass and Dicraptoris Dicholoma, while fat content was highest in those grown on Giant Juncao Grass and Dicraptoris Dicholoma, fol-lowed by Wasted Tea Leaves and Sawdust. Protein content was significantly higher in fruiting bodies cultivated on Wasted Tea Leaves and Sawdust, followed by Cottonseed Hulls. Carbon dioxide emissions varied across substrates, with the highest emissions observed during the maturity stage of the fruiting bodies grown on Giant Juncao Grass and Dicraptoris Dicholoma, and Wasted Tea Leaves and Sawdust. Emissions decreased sharply 110 days after cultiva-tion. Essential metabolites such as dopamine and caffeine were enriched in fruiting bodies grown on Wasted Tea Leaves, Sawdust, and Cottonseed Hulls. In contrast, tyramine and uracil were enriched in those grown on Cotton-seed Hulls.

The structure characterization of two new polysaccharides from Agrocybe aegerita (AA-P) and Hygrophorus olivaceoalbus (HO-P) by HPGPC, GC-MS, NMR and FT-IR indicated that AA-P was composed of Galactose, Glucose and Arabinose, in the ratio of 3:2:1. Its skeleton structure was consisted of (1→4)-Arap, (1→4,6)-Glup and (1→6)-Galp with one branched chain. The HO-P was consisted of mannose, galactose and glucose in a ratio of 1:1:2. Its skeleton structure was consisted of (1→6)-Galactose residues, (→1)-glucose residues, (1→4)-glucose residues and (1→4,6)-D-mannose residues. There were two branched chains connected to the main chain. AA-P and HO-P had the best stimulation effect on B cells and RAW264.7 cells, respectively, and could both mainly by impacting and reducing G0/G1 phase which lead to a significant proliferation of B cells, T cells and RAW264.7 cells. In addition, AA-P and HO-P could significantly promote the secretion of TNF-α from T cells, the secretion of IgA, IgD, IgE, IgG and IgM from B cells, and the secretion of TNF-α from RAW264.7 cells, but neither of them could impact the secretion of IL-1β from RAW264.7 cells.