Fungi occupy important environmental, cultural, and socioeconomic roles. However, biological research of this diverse kingdom has lagged behind that of other phylogenetic groups. This is partially the result of the notorious difficulty in culturing a diverse array of filamentous fungal species due to their (i) often unpredictable growth, (ii) unknown preferences for culturing conditions, and (iii) long incubation times compared with other microorganisms such as bacteria and yeasts. Given the complexity associated with concurrently culturing diverse fungal species, developing practical methods for preserving as many species as possible for future research is vital. The widely accepted best practice for preserving fungal tissue is the use of cryogenic biobanking at -165 C, allowing for the preservation and documentation of stable genetic lineages, thus enabling long-term diversity-centered research. Despite the extensive literature on fungal cryopreservation, substantial barriers remain for implementation of cryogenic biobanks in smaller mycological laboratories. In this work, we present practical considerations for the establishment of a fungal culture biobank, as well as provide evidence for the viability of 61 fungal genera in cryogenic storage. By providing a pragmatic methodology for cryogenically preserving and managing many filamentous fungi, we show that creating a biobank can be economical for independently owned and operated mycology laboratories, which can serve as a long-term resource for biodiversity, conservation, and strain maintenance.

Hericium erinaceus is a medicinal mushroom valued in the wellness industry for its neuroprotective, immunomodulatory, and antioxidant activities. While many extracts and bioactive compounds from both mycelium and fruit bodies have been characterized, the mechanisms driving their effects are not fully understood. Here, the transcriptomic and protein-level effects of H. erinaceus mycelium (HDLM) in human peripheral blood mononuclear cells (PBMCs) were investigated, along with antioxidant and iron chelating activity. A commercially available H. erinaceus fruit body extract (FBE) claiming high β-glucan content was included in a subset of assays to compare immune-related outcomes between mycelial and fruit body constituents. HDLM activated a wide array of immune- and oxidative stress-related transcripts and pathways, exhibited significant antioxidant activity, and consistently reduced IL-1β, TNF-α, and IL-8 during LPS challenge while maintaining low basal cytokine expression, indicating targeted immunomodulatory activity. FBE almost doubled production of IL-1β when challenged by LPS, whereas HDLM significantly decreased production of this stress mediator. HDLM also demonstrated augmented iron chelating ability when compared to FBE. Depending on tissue source and preparation methods, different H. erinaceus materials may either potentiate or quench stress responses, highlighting the need for further bioactivity and safety comparisons across H. erinaceus supplements, particularly with respect to cytokine regulation under conditions of immune challenge.

Agarikon (Laricifomes officinalis syn. Fomitopsis officinalis) is an endangered fungus belonging to a unique lineage in the Polyporales (Basidiomycota) with a growing body of evidence supporting its medicinal value. In this study, we report the hybrid de novo assembly and annotation of the nuclear and mitochondrial genomes of L. officinalis, with a nuclear genome size of 28.76 Mb assembled across 66 contigs (51.96% GC content; BUSCO completeness of 99.4%) and a complete core mitochondrial genome size of 197.67 kb. Structural and functional annotation of the nuclear genome yielded 8,717 predicted genes including 8,604 protein-coding genes, with 287 genes in 27 candidate biosynthetic gene clusters. We characterized the mating type loci matA and matB, consistent with a tetrapolar mating system, and identified genes encoding key enzymes involved in triterpenoid and polyketide biosynthetic pathways that lead to the production of a diverse array of secondary metabolites. Additionally, we conducted maximum likelihood phylogenomic analysis to confirm the taxonomic position of L. officinalis among other species in the Polyporales using protein sequences for 730 shared BUSCO genes. This high-quality annotated genome of L. officinalis will serve as a foundation for further investigations into its evolutionary history, provide a reference for future population genomic analyses, and elucidate mechanisms underlying the synthesis of the bioactive compounds responsible for agarikon's wide-ranging medicinal benefits.

Agarikon ( Laricifomes officinalis syn. Fomitopsis officinalis ) is an endangered fungus belonging to a unique lineage in the Polyporales (Basidiomycota) with a growing body of evidence supporting its medicinal value. In this study, we report the hybrid de novo assembly and annotation of the first L. officinalis nuclear and mitochondrial genome sequences, with a nuclear genome size of 28.76 Mb assembled across 66 scaffolds (51.96% GC content; BUSCO completeness of 99.4%), and a complete core mitochondrial genome size of 197.67 kb. Structural and functional annotation of the nuclear genome yielded 8,717 predicted genes including 8,604 protein-coding genes, with 310 genes in 27 biosynthetic gene clusters. We characterized the mating type loci matA and matB , consistent with a tetrapolar mating system, and identified genes encoding key enzymes involved in triterpenoid and polyketide biosynthetic pathways that lead to the production of a diverse array of secondary metabolites. Additionally, we conducted maximum likelihood phylogenomic analysis to confirm the taxonomic position of L. officinalis among 21 species in Polyporales using protein sequences for 860 shared BUSCO genes. This high-quality annotated genome of L. officinalis will serve as a foundation for further investigations into the evolutionary history of this distinct fungal lineage, provide a reference for future population genomic analyses, and elucidate mechanisms underlying the synthesis of the bioactive compounds responsible for agarikon′s wide-ranging medicinal benefits.

Hericium erinaceus is a medicinal mushroom valued for its neuroprotective, immunomodulatory, and antioxidant activities. While many extracts and bioactive compounds from both mycelium and fruit bodies have been characterized, the mechanisms driving their effects are not fully understood. Here, the transcriptomic and protein-level effects of H. erinaceus mycelium (HDLM) in human peripheral blood mononuclear cells (PBMCs) were investigated, along with antioxidant and iron chelating activity. A commercially available β-glucan-enriched H. erinaceus fruit body extract (FBE) was included in a subset of assays to compare immune-related outcomes between mycelial and fruit body constituents. HDLM activated a wide array of immune- and oxidative stress–related transcripts and pathways, exhibited significant antioxidant activity, and consistently reduced IL-1β, TNF-α, and IL-8 during LPS challenge while maintaining low basal cytokine expression, indicating targeted immunomodulatory activity. FBE almost doubled production of IL-1β when challenged by LPS, whereas HDLM significantly decreased production of this stress mediator. HDLM also demonstrated augmented iron chelating ability when compared to FBE. Depending on tissue source and preparation methods, different H. erinaceus materials may either potentiate or quench stress responses, highlighting the need for bioactivity and safety comparisons across H. erinaceus supplements.

Agarikon ( Fomitopsis officinalis , syn. Laricifomes officinalis ) is a fungus with millennia of traditional use across many cultures with modern research supporting the antimicrobial, antiviral, anticancer, antioxidant, and immune-modulating properties of both mycelium and fruit body. Due to the slow growth and old-growth forest habitat of agarikon fruit bodies, its mycelium represents an easily cultivated immunomodulating and stress buffering preparation, underscored by recent clinical trials of a blend of agarikon and Trametes versicolor mycelium. We investigated the transcriptomic effects of agarikon mycelium in human peripheral blood mononuclear cells (PBMCs) under both basal and LPS-stimulated conditions, alongside evaluations of antioxidant, iron chelating, and kinase binding activity. Two agarikon fractions were also assessed for effects on cell viability and proliferation and induction of select cytokine targets. Under basal conditions, agarikon mycelium selectively engaged the innate immune response through IL-1 and NF-κB axes, balanced by increases in anti-inflammatory mediators such as IL-1RA and decreases in toll-like receptor transcripts. Under LPS-stimulated conditions, this innate immune response was modified, with measured increases in immune effectors (including TLR5 ) observed in response to induced stress, alongside accompanying transcript decreases in cytokine pathways that can overstimulate the immune system. Overall, agarikon mycelium demonstrated a coordinated, context-dependent immune response profile, supporting its stress-buffering and immune-modulating potential and warranting continued clinical validation.