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Abstract BackgroundTruffles are subterranean fungal fruiting bodies that are highly prized for their culinary value. Cultivation of truffles was pioneered in Europe and has been successfully adapted in temperate regions throughout the globe. Truffle orchards have been established in North America since the 1980s, and while some are productive, there are still many challenges that must be overcome to develop a viable North American truffle industry. These challenges include extended delays between establishment and production, comparatively low yields, high spatial heterogeneity in yield distribution, and orchard contamination with lower-value truffle fungi. AimHere we review known requirements for truffle production including necessary environmental conditions, reproductive biology, and effective agronomic practices. ContentWe consider the potential limitations of importing exotic host-fungal associations into North America where there is already a rich community of competing ectomycorrhizal fungi, host pests and pathogens. We also describe the status of the North American truffle industry with respect to market potential, including production costs, pricing, and biological and socioeconomic risk factors. A critical aspect of modern trufficulture involves monitoring with genetic tools that supply information on identity, abundance and distribution of fungal symbionts, abundance of competitive and contaminating fungi, and insight into the interactions between fungal mating types that are fundamental to the formation of truffle primordia. ImplicationsCultivation of the ectomycorrhizal truffle symbiosis requires application of pragmatic agronomic practices, adopting rigorous quality control standards, and an understanding of fungal biology, microbiology, and molecular biology. Consequently, significant interdisciplinary collaboration is crucial to further develop the North American truffle industry. 

Abstract The overarching goal of this impact project is to make mycology accessible to more agriscience educators and students. Lesson plans were prepared to link core competencies and science standards to the Wild‐Foraged Mushroom certification. Incorporating mycology into the classroom has many benefits, including discussions on food safety and regulation, the role of ecology in agroecosystems, and taxonomic identification skills. Fungi also play many different roles in the ecosystem, including decomposers, mutualists, and parasites. Lesson plans in three topic areas were produced: mushroom identification and fungal ecology, mushroom growth and food safety, and mushrooms as a renewable resource. Examples of hands‐on learning and connections to the Wild‐Foraged Mushroom certification are provided. This certification is available in the state of Michigan; however, lessons could be adapted for use in other regions of the United States. Looking at taxonomy, ecology, food science, and economics through the lens of mycology is an engaging way to motivate students while potentially helping them earn a certification. 

Societal Impact Statement Morels (Morchellaspp.) are specialty mushrooms that fetch high prices from wild‐foraged or indoor grown suppliers. Outdoor cultivation could expand availability and diversify morel crops. Participatory research trials in the United States during 2021–2023 resulted in low, uneven yields. Cost accounting reveals that in 2023, a producer needed to achieve an average morel yield of 0.16 lb/ft of row to break even. This threshold was sensitive to prices and labor costs. While these findings are preliminary due to a small sample and experimental conditions, they establish baseline indicators for the yields needed for outdoor morel cultivation to break even financially. 

Abstract Truffle growers devote great efforts to improve black truffle productivity, developing agronomic practices such as ‘truffle nests’ (peat amendments that are supplemented with truffle spore inoculum). It has been hypothesized that improved fruiting associated with nests is linked to stimulation of truffle mycelia previously established in soil or to changes generated in soil fungal community. To assess this, we used real-time PCR to quantify black truffle extraradical mycelium during 2 years after nests installation. We also characterized the fungal community via high-throughput amplicon sequencing of the ITS region of rRNA genes. We found that neither the abundance of truffle mycelium in nests nor in the soil—nest interphase was higher than in the bulk soil, which indicates that nests do not improve mycelial growth. The fungal community in nests showed lower richness and Shannon index and was compositionally different from that of soil, which suggests that nests may act as an open niche for fungal colonization that facilitates truffle fruiting. The ectomycorrhizal fungal community showed lower richness in nests. However, no negative relationships between amount of truffle mycelium and reads of other ectomycorrhizal fungi were found, thus countering the hypothesis that ectomycorrhizal competition plays a role in the nest effect. 

Morchellaspecies have considerable significance in terrestrial ecosystems, exhibiting a range of ecological lifestyles along the saprotrophism-to-symbiosis continuum. However, the mitochondrial genomes of these ascomycetous fungi have not been thoroughly studied, thereby impeding a comprehensive understanding of their genetic makeup and ecological role. In this study, we analysed the mitogenomes of 30Morchellaceaespecies, including yellow, black, blushing and false morels. These mitogenomes are either circular or linear DNA molecules with lengths ranging from 217 to 565 kbp and GC content ranging from 38% to 48%. Fifteen core protein-coding genes, 28–37tRNAgenes and 3–8rRNAgenes were identified in theseMorchellaceaemitogenomes. The gene order demonstrated a high level of conservation, with thecox1gene consistently positioned adjacent to thernSgene andcobgene flanked byaptgenes. Some exceptions were observed, such as the rearrangement ofatp6andrps3inMorchella importunaand the reversed order ofatp6andatp8in certain morel mitogenomes. However, the arrangement of thetRNAgenes remains conserved. We additionally investigated the distribution and phylogeny of homing endonuclease genes (HEGs) of the LAGLIDADG (LAGs) and GIY-YIG (GIYs) families. A total of 925 LAG and GIY sequences were detected, with individual species containing 19–48HEGs. These HEGs were primarily located in thecox1,cob,cox2andnad5introns and their presence and distribution displayed significant diversity amongst morel species. These elements significantly contribute to shaping their mitogenome diversity. Overall, this study provides novel insights into the phylogeny and evolution of theMorchellaceae. 

Through their expansive mycelium network, soil fungi alter the physical arrangement and chemical composition of their local environment. This can significantly impact bacterial distribution and nutrient transport and can play a dramatic role in shaping the rhizosphere around a developing plant. However, direct observation and quantitation of such behaviors is extremely difficult due to the opacity and complex porosity of the soil microenvironment. In this study, we demonstrate the development and use of an engineered microhabitat to visualize fungal growth in response to varied levels of confinement. Microfluidics were fabricated using photolithography and conventional soft lithography, assembled onto glass slides, and prepared to accommodate fungal cultures. Selected fungal strains across three phyla (Ascomycota:Morchella sextalata,Fusarium falciforme; Mucoromycota:Linnemannia elongata,Podila minutissima,Benniella; Basidiomycota:Laccaria bicolor, andSerendipitasp.) were cultured within microhabitats and imaged using time-lapse microscopy to visualize development at the mycelial level. Fungal hyphae of each strain were imaged as they penetrated through microchannels with well-defined pore dimensions. The hyphal penetration rates through the microchannels were quantified via image analysis. Other behaviors, including differences in the degree of branching, peer movement, and tip strength were also recorded for each strain. Our results provide a repeatable and easy-to-use approach for culturing fungi within a microfluidics platform and for visualizing the impact of confinement on hyphal growth and other fungal behaviors pertinent to their remodeling of the underground environment. 

Cistaceae are shrubs, subshrubs and herbs that often occur in stressful, fire-prone or disturbed environments and form ectomycorrhizal (ECM) associations with symbiotic fungi. Although some Cistaceae are long-lived shrubs that grow to significant size, others are herbaceous annuals or short-lived plants. Thus, Cistaceae are atypical ECM hosts that are fundamentally different in their biology from trees that are the more typically studied ECM hosts. The Mediterranean region is the center of diversity for Cistaceae and the ectomycorrhizal fungi associated with Cistaceae hosts have primarily been studied in Europe, North Africa, and the Middle East. Mediterranean Cistaceae often host diverse communities of ECM fungi, but they also act as hosts for some ECM fungi that putatively show host-specificity or strong host preference for Cistaceae (including species of Delastria, Hebeloma, Terfezia, and Tirmania). The ECM associations of Cistaceae in North America, however, remain highly understudied. Here we use fungal DNA metabarcoding to document the ectomycorrhizal fungal communities associated with Crocanthemum and Lechea (Cistaceae) in open, fire-prone sandhill habitats in north Florida. At each site we also sampled nearby Pinus to determine whether small, herbaceous Cistaceae have specialized ECM fungi or whether they share their ECM fungal community with nearby pines. The ECM communities of Florida Cistaceae are dominated by Cenococcum (Ascomycota) and Russula (Basidiomycota) species but were also significantly associated with Delastria, an understudied genus of mostly truffle-like Pezizales (Ascomycota). Although many Cistaceae ECM fungi were shared with neighboring pines, the ECM communities with Cistaceae were nonetheless significantly different than those of pines.