This study presents the first tissue-resolved targeted metabolomic analysis of artificially cultivated Termitomyces upsilocystidiatus fruiting bodies using LC-MS/MS. We identified pronounced metabolic divergence between the pileus and stipe. The pileus was enriched in a nitrogen-recycling and antioxidant module, exemplified by L-citrulline (~13.5-fold higher than stipe, p < 0.01) and urea, while the stipe accumulates sulfur-derived and oxidized metabolites such as L-homocystine (~3.5-fold higher, p < 0.01) and methionine sulfoxide. Lipid profiles further distinguished the two tissues: the pileus featured high levels of linoleic acid-derived oxylipins, including 13(S)-HODE and 12(13)-DiHOME (~9.7-fold and ~303-fold enrichment, respectively, p < 0.01), suggesting a role in signaling and redox buffering. In contrast, the stipe preferentially accumulated oxidized eicosanoids (e.g., 5-oxoETE) and thromboxane B1, indicative of a stress-responsive lipid network. Together, these metabolite-level observations support a tentative “pileus-synthesis/stipe-defense” dual-hub model. This work provides a quantitative metabolic framework for understanding tissue specialization in a symbiotic fungus and offers practical entry points for cultivation optimization and quality control of Termitomyces.

This study combines Oxford Nanopore (ONT) third-generation sequencing with targeted amino acid metabolomics to elucidate the mechanisms underlying the structural and metabolic responses of the microbial community in Golden Ear Mushroom (Naematelia sinensis) during Trichoderma infection. By comparing healthy tissue (MOCK), adjacent healthy areas (HAF) and the core lesion area (DiR), the results indicate that pathogen infection significantly reduces bacterial community diversity, with a progressive decline observed across these regions. In the DiR region, the fungal community underwent significant restructuring, with the abundance of the Trichoderma genus (T. lixii and T. afroharzianum) rising to over 45%, whilst that of host symbiotic fungi (Stereum and Tremella) decreased by 50–60%. Metabolomic analysis indicated that levels of various amino acids and antioxidant-related metabolites were significantly reduced in the host tissue of the DiR region, suggesting that amino acid metabolism was inhibited. Concurrently, changes were observed in certain metabolites associated with nitrogen metabolism (e.g., L-glutamine). KEGG analysis further revealed that amino acid biosynthesis and D-amino acid metabolic pathways were inhibited, whilst ABC transporters and arginine/proline metabolic pathways were activated. All metabolic changes originated from the host fungal tissue itself, rather than from commensal microorganisms. In summary, Trichoderma may promote the infection process by disrupting the host microbial community and metabolic networks, providing a theoretical basis for understanding the mechanisms of fungal diseases and their control.

Armillaria is a genus of macrofungi with high ecological, biological, medicinal, and edible value. As facultative plant pathogens and nutritional symbionts, Armillaria species support the growth of valuable medicinal plants including Gastrodia elata and Polyporus umbellatus. They also exhibit unique traits such as exceptional longevity, widespread clonal expansion, rhizomorph formation, and bioluminescence, making them a valuable model for studying fungal ecology, symbiosis, specialized metabolism, and applied research. This review summarizes recent progress in Armillaria research, covering biological characteristics, nutritional components, bioactive constituents, species identification, genomic resources, and biosynthetic pathways. We discuss advances in artificial cultivation and the regulatory roles of exogenous phytohormones in mycelial and rhizomorph development. The nutritional value of fruiting bodies is highlighted, with a focus on key pharmacologically active metabolites such as protoilludane-type sesquiterpenes and polysaccharides. We also review multilocus phylogenetic analysis, comparative genomics, and the biosynthetic gene clusters of melleolides and bioluminescence, which have improved understanding of Armillaria evolution and functional differentiation.