Puffball Research Articles

An integrative taxonomic and molecular identification of Calvatia holothurioides (Lycoperdaceae): the present status of genus Calvatia in India

Genus Calvatia belongs to a gasteroid fungi which are commonly known as a puffball mushrooms including around 47 species worldwide. Approximately 25 species of the genus Calvatia have been reported previously in India from various forest regions; however, only 15 species of these are accepted on their current taxonomic status because several of them are synonymized with other genera or species. The present study reports the distribution of Calvatia holothurioides which was collected from Jambughoda Wildlife Sanctuary in Gujarat state, India. The identification was carried out based on the morphological characteristics as well as molecular phylogenetic analysis using nuclear rDNA Internal Transcribed Spacer (ITS) and rDNA LSU gene sequences data. Nucleotide sequences of different gene loci (ITS and LSU) were submitted into the BOLD data system for DNA barcoding. Additionally, this study also provides a literature-based checklist for the present status of Calvatia species reported in India so far.

Medicinal Potential of the Giant Puffball Mushroom Calvatia gigantea (Agaricomycetes): A Review.

Calvatia gigantea, commonly known as the giant puffball mushroom, has traditionally been regarded as a significant edible and medicinal species due to its wide spectrum of bioactive compounds and its health-promoting properties. This study aims to systematize the knowledge on the nutritional value and therapeutic potential of C. gigantea, highlighting its role in traditional and contemporary medicine. The mushroom is recognized for its nutritional content, including easily digestible protein, carbohydrates, fiber, phenolic compounds, vitamins, and minerals, while being low in calories, cholesterol, and sodium. Furthermore, C. gigantea exhibits a range of biological effects, such as antioxidant, anticancer, antimicrobial, antidiabetic, and wound-healing properties, attributed to its diverse chemical composition that includes unsaturated fatty acids, free amino acids, polysaccharides, and bioactive metabolites.

Exploring the Bioactive Potential of Calostoma insigne, an Endangered Culinary Puffball Mushroom, from Northeastern Thailand.

Calostoma insigne puffball mushrooms are only found in forests with rich biodiversity in very few countries including Thailand, and their biofunctions remain largely unexplored. This study used the agar disk diffusion assay, the anti-glucosidase assay, and the 3, 4, 5-dimethylthiazol-2-yl-2-5-diphenyltetrazolium bromide (MTT) assay to evaluate the bioactive potential of these endangered puffball mushrooms. Internal transcribed spacer (ITS) gene analysis identified C. insigne, a puffball mushroom with green, globose, and spiny spores. Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the polysaccharide structure while scanning electron microscopy (SEM) revealed a fiber-like network. The ethanolic gelatinous fruiting body extract exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging capacity (57.96%), a ferric ion-reducing antioxidant power (FRAP) value of 1.73 mg FeSO4/g, and α-glucosidase inhibition (73.18%). C. insigne cytotoxicity was effective towards HT-29 colon cancer cells using the MTT assay (IC50 of 770.6 µg/mL at 72 h) and also showed antiproliferative capacity (IC50 of 297.1 µg/mL). This puffball mushroom stimulated apoptotic genes and proteins (caspase-3, Bax, and p21) via an intrinsic apoptotic pathway in HT-29 cells. In the laboratory, the medium formula consisting of 20% potato, 2% sucrose, and 0.2% peptone was optimal to increase fungal mycelial biomass (2.74 g DW/100 mL), with propagation at pH 5.0 and 30 °C. Puffball mushrooms are consumed as local foods and also confer several potential health benefits, making them worthy of conservation for sustainable utilization.

Study of stable sodium ion storage in porous carbon derived from puffball biomass

The pursuit of affordable anode materials for sodium-ion batteries (SIBs) has become imperative in light of the rising energy demand in modern society. Hard carbon (HC), a highly promising candidate for SIBs, has garnered interest among researchers. In this study, we present a novel approach involving the thermal treatment of low-cost puffball (PB) at various temperatures to produce cost-effective porous carbon, which holds great potential as an anode material for SIBs. The carbon derived from the treated puffball biomass showcases distinctive morphological features, including carbon tubes and plentiful balloon-like porous structures. This morphological structure can prevent the agglomeration and stacking of carbon materials, which can effectively increase the active sites and energy storage channels. The resulting puffball biomass-derived carbon exhibits a reversible capacity of 205.05 mAh g−1 at 100 mA g−1. Even after 600 cycles at a current density of 2 A g−1, the discharge capacity remains as high as 146.98 mAh g−1. These findings emphasize the remarkable performance and cycling stability of the carbon material. Significantly, the puffball biomass-derived carbon exhibited an impressive initial Coulomb efficiency of 57.6%, which is a noteworthy achievement. Moreover, we employed in situ XRD to investigate its energy storage mechanism in sodium-ion batteries. When integrated into a complete cell with Na3V2(PO4)3 (NVP) and bulking materials, the carbon anode exhibited impressive multiplicative performance and maintained good cycling stability. These findings not only provide new insights into the design of carbon anodes for sodium-ion batteries but also emphasize their exceptional attributes such as excellent initial coulombic efficiency, impressive rate capability, and cost-effectiveness for sodium storage.

Gautieria mianjin (Gomphales, Basidiomycota), a new species of false truffle from China

A new species of false truffle, Gautieria mianjin, collected from northwestern China, is described based on morphological and phylogenetic evidence. It is characterized by its irregularly subglobose basidioma in the size of 2.5–5 cm in diameter with brain-like to honeycomb-like gleba and subglobose to broadly ellipsoid basidiospores measured 11–14×10–12 μm with 10–14 longitudinal ribs.

On the origin of feces: Fungal diversity, distribution, and conservation implications from feces of small mammals

AbstractFungi are highly diverse, but only a small fraction of the total estimated species have been characterized. Often, the extent of diversity and distribution of fungal communities is difficult or near impossible to assess due to the fact that many fungi are cryptic and persist predominantly hidden within substrates such as soil or plant material. This is particularly true for hypogeous sporocarps, such as truffles and false truffles, which are extremely difficult to survey in a systematic manner. However, hypogeous fungi have evolved traits that make them highly attractive to animals, such as small mammals, which ingest and disperse fungal spores through defecation. Here, samples of feces from 138 small mammal museum vouchers collected in the western United States were assessed for total fungal diversity using a dual‐index metabarcoding approach. Our findings exhibit many identifications within the mushroom‐forming fungi (Agaricomycetidae), with 65 of the 138 samples containing sequences belonging to several species of the false truffle‐forming genus Rhizopogon. Metadata for each collection, such as geospatial coordinates, can be used as a proxy for the presence or absence of Rhizopogon species identified in their feces. Utilizing these proxy data from only a few years of sampling, these records quadrupled the rate of observations of Rhizopogon made over the past 100 years, including some species that have only been recorded once previously. This substantial increase in datapoints has implications for how fungal distributions are interpreted, with direct impact on standard assessment tools for fungal conservation.

Hybrid material based on subgleba of mosaic puffball mushroom (Handkea utriformis) as an adsorbent for heavy metal removal from aqueous solutions

The alkali treated subglebal tissue of the mosaic puffball (Handkea utriformis) (Sa) and Sa modified with hydroxyapatite (Sa-HAp), obtained by successive ionic layer adsorption and reaction (SILAR) method, were used for the removal of Pb2+, Cd2+ and Ni2+ from aqueous solution. The materials were characterized by FT-IR, Raman, SEM and EDS analysis and by determination of pHPZC. The adsorption performances of Sa and Sa-HAp were assessed in batch experiments at different pH, contact times, temperatures and mass of the adsorbent. Different models of adsorption isotherms were used, and the best fit was obtained with the Langmuir model. Maximum adsorption capacities of Sa towards Pb2+, Cd2+ and Ni2+ were 44.82, 15.54 and 17.21 mg g−1, while for Sa-HAp were 79.55, 52.59 and 45.01 mg g−1, respectively. Kinetic data were well fitted by a pseudo second-order model, while thermodynamic studies disclose spontaneous and endothermic adsorption process. The Sa-Hap was successfully regenerated with 1 M NaCl and after the fifth desorption cycle and 10 h achieved 82.9, 69.7 and 60.4 %, while for 0.5 M NaCl + 0.5 M NaOH and 1 h was 78.3, 64.1, 57.5 % of desorbed Pb2+, Cd2+ and Ni2+, respectively. The competitive study and results from a column system confirmed good applicability of Sa-HAp adsorbent.

An Overview of 24 Years of Molecular Phylogenetic Studies in Phallales (Basidiomycota) With Notes on Systematics, Geographic Distribution, Lifestyle, and Edibility.

The order Phallales (Basidiomycota) is represented by gasteroid fungi with expanded and sequestrate basidiomata, known as stinkhorns and false truffles. In phalloids, the first DNA sequence was published in 1997, and after that, some studies aimed to resolve phylogenetic conflicts and propose new species based on DNA markers; however, the number of families and genera in the order still generates controversies among researchers. Thus, this work aims to provide an overview of Phallales diversity represented by selected DNA markers available in public databases. We retrieved Phallales sequences from DNA databases (GenBank and UNITE) of seven markers: ITS (internal transcribed spacer), nuc-LSU (nuclear large subunit rDNA), nuc-SSU (nuclear small subunit rDNA), mt-SSU (mitochondrial small subunit rDNA), ATP6 (ATPase subunit 6), RPB2 (nuclear protein-coding second largest subunit of RNA polymerase), and TEF1-α (translation elongation factor subunit 1α). To compose our final dataset, all ITS sequences retrieved were subjected to BLASTn searches to identify additional ITS sequences not classified as Phallales. Phylogenetic analyses based on Bayesian and maximum likelihood approaches using single and combined markers were conducted. All ITS sequences were clustered with a cutoff of 98% in order to maximize the number of species hypotheses. The geographic origin of sequences was retrieved, as well as additional information on species lifestyle and edibility. We obtained a total of 1,149 sequences, representing 664 individuals. Sequences of 41 individuals were unidentified at genus level and were assigned to five distinct families. We recognize seven families and 22 genera in Phallales, although the delimitation of some genera must be further revisited in order to recognize only monophyletic groups. Many inconsistencies in species identification are discussed, and the positioning of genera in each family is shown. The clustering revealed 118 species hypotheses, meaning that approximately 20% of all described species in Phallales have DNA sequences available. Information related to geographic distribution represents 462 individuals distributed in 46 countries on all continents, except Antarctica. Most genera are saprotrophic with only one putative ectomycorrhizal genus, and 2.1% of the legitimate specific names recognized in Phallales are confirmed edible species. Great progress in the molecular analyses of phalloids has already been made over these years, but it is still necessary to solve some taxonomic inconsistencies, mainly at genus level, and generate new data to expand knowledge of the group.

Evaluating mercury concentrations in edible plant and fungi species in the Canadian Arctic environment.

Levels of environmental mercury (Hg) within the Canadian Arctic are a current area of concern. Although efforts have been made to reduce Hg released into the environment, levels remain elevated in flora and fauna. This study examined the concentrations of Hg in soil and naturally occurring edible plant and fungi species, identified by local Inuit residents, from eight locations in Iqaluit, Nunavut, and the surrounding area during the summers of 2018 and 2019. Total Hg concentrations were obtained in 24 soil samples, 112 flora samples from 23 plant and five lichen species, and 157 fungal samples from eight species. Median Hg concentrations in plant species ranged from 0.005μg g-1 Hg dry weight (dw) in Saxifraga cernua to 0.19μg g-1 Hg dw in Oxytropis maydelliana. Median concentrations in edible fungi species ranged from 0.084μg g-1 Hg dw in the Cortinarius croceus (non-puffball species) to 1.6μg g-1 Hg dw in Lycoperdon perlatum (a puffball mushroom). Additionally, median Hg concentration in puffball species (1.4μg g-1 ) were higher than non-puffball species (0.12μg g-1 ). Three puffball species were assessed for methylmercury (MeHg), with mean concentrations ranging from 0.013 to 0.085μg g-1 MeHg dw. Limited research has been conducted on Hg uptake in naturally occurring edible plant and fungi species of the Canadian Arctic. This study contributes important information on Hg accumulation and processes in edible plant and fungi Arctic species, is the first to focus on plants used by the local Indigenous community, and demonstrates a need for further studies to assess Hg in Arctic environments.

New ribosome-inactivating proteins and other proteins with protein synthesis-inhibiting activities.

Ribosome-inactivating proteins (RIPs) consist of three varieties. Type 1 RIPs are single-chained and approximately 30-kDa in molecular weight. Type 2 RIPs are double-chained and composed of a type 1 RIP chain and a lectin chain. Type III RIPs, such as maize b-32 barley and JIP60 which are produced as single-domain proenzymes, possess an N-terminal domain corresponding to the A domain of RIPs and fused to a C-terminal domain. In addition to the aforementioned three types of RIPs originating from flowering plants, there are recently discovered proteins and peptides with ribosome-inactivating and protein synthesis inhibitory activities but which are endowed with characteristics such as molecular weights distinctive from those of the regular RIPs. These new/unusual RIPs discussed in the present review encompass metazoan RIPs from Anopheles and Culex mosquitos, antimicrobial peptides derived from RIP of the pokeweed Phytolacca dioica, maize RIP (a type III RIP derived from a precursor form), RIPs from the garden pea and the kelp. In addition, RIPs with a molecular weight smaller than those of regular type 1 RIPs are produced by plants in the Cucurbitaceae family including the bitter gourd, bottle gourd, sponge gourd, ridge gourd, wax gourd, hairy gourd, pumpkin, and Chinese cucumber. A small type II RIP from camphor tree (Cinnamomum camphora) seeds and a snake gourd type II RIP with its catalytic chain cleaved into two have been reported. RIPs produced from mushrooms including the golden needle mushroom, king tuber mushroom, straw mushroom, and puffball mushroom are also discussed in addition to a type II RIP from the mushroom Polyporus umbellatus. Bacterial (Spiroplasma) RIPs associated with the fruitfly, Shiga toxin, and Streptomyces coelicolor RIP are also dealt with. The aforementioned proteins display a diversity of molecular weights, amino acid sequences, and mechanisms of action. Some of them are endowed with exploitable antipathogenic activities.

Cumulative Effects of Constituents from the Mushroom Calvatia nipponica on the Contractility of Penile Corpus Cavernosum Smooth Muscle

Calvatia nipponica, a puffball mushroom (Agaricaceae), is thought to be an aphrodisiac, as this mushroom is traditionally known to improve sexual function in males. As part of the systematic study to determine the bioactive secondary metabolites from C. nipponica responsible for aphrodisiac effects, chemical analysis of methanol (MeOH) extracts of the fruiting bodies of C. nipponica resulted in the isolation of two major compounds: N,N-dimethyl-anthranilic acid (1) and (7Z,10Z)-7,10-octadecadienoic acid methyl ester (2). Compounds 1 and 2 were evaluated for cumulative dose-dependent relaxation responses to precontracted penile corpus smooth muscle (PCCSM). Results show that compounds 1 and 2 exhibited a maximum relaxation effect of 20.33 ± 2.18% and 24.63 ± 3.60%, respectively. These findings indicate that compounds 1 and 2, major components of C. nipponica, could potentially be used to treat erectile dysfunction, functioning as natural aphrodisiacs.

281. Hypersensitivity Pneumonitis Diagnosed with Broad-Range PCR Testing after Exposure to Battarrea Mushroom Spores

Background Battarrea puffball mushrooms are found extensively worldwide and contain spore-containing sacs. Inhalation of the spores of similar mushrooms, such as Lycoperdon, have been implicated in cases of lycoperdonosis—a syndrome of hypersensitivity pneumonitis. We report a case of hypersensitivity pneumonitis confirmed to be secondary to Battarrea spore exposure diagnosed by broad-range PCR.MethodsA 23-year-old homeless man with a history of methamphetamine use presented to the emergency department with a 2-week history of fevers, chills, productive cough, and malaise. He reported his symptoms began soon after eating a long-stemmed mushroom he found growing next to a building. He reported inhaling particles from the mushroom when he picked it up prior to eating it. He vomited within 1 hour of ingestion, and then had a worsening progression of cough and malaise over the following 2 weeks. In the emergency department, he was noted to have leukocytosis and mild elevation of transaminases. He required supplemental oxygen due to hypoxemia. CT scan of his chest demonstrated extensive bilateral nodular pulmonary infiltrates. He was admitted and started on treatment for community-acquired pneumonia. Over the next several days, he had worsening respiratory failure, and routine work up for infectious etiologies was unrevealing. To further investigate, bronchoscopy and bronchoalveolar lavage (BAL) was performed and routine bacterial, fungal and mycobacterial cultures and cytology with Gomori Methanamine-silver and acid-fast stains were negative. BAL fluid was sent for broad range DNA testing by PCR. Antibiotic therapy was stopped, and he was started on steroids to treat presumed hypersensitivity pneumonitis. He recovered rapidly and was discharged on a course of oral corticosteroids.ResultsAfter the patient was discharged, molecular testing of BAL fluid resulted with detection of Battarrea species DNA using 28s and ITS primer sets. DNA from no other pathogens was detected.ConclusionIdentified through broad range DNA PCR testing, exposure to Battarrea mushroom spores may be a previously unreported cause of hypersensitivity pneumonitis. PCR testing should be considered in the workup of hypersensitivity pneumonitis with known or suspected exposure to mushroom spores. Disclosures All authors: No reported disclosures.

Molecular identification, in vivo and in vitro activities of Calvatia gigantea (macro-fungus) as an antidiabetic agent

ABSTRACT Mushrooms are cherished as sources of food, nutrients and medicine. Inadequate data on the identity and medicinal properties of many wild Nigerian mushrooms has limited their utilization. This work was carried out to identify and authenticate a puffball mushroom using molecular tools and investigate its antidiabetic properties. Taxonomic guides were employed in morphological identifying the mushroom as Lycoperdon umbrinum, methanol extract of fruiting bodies was evaluated for antidiabetic activity using in vitro α-amylase assay and in vivo activity in the alloxan-induced diabetic rat model. The macro fungus was identified using Internal Transcribed Spacers (ITS) sequence analysis after which sequences generated were compared using the basic local alignment search tool (BLAST) at NCBI GenBank. In the acute in vivo test, the 400 mg/kg dose showed the best activity with percentage reduction in blood glucose 29.3%, compared with 5 mg/kg glibenclamide at 15%. The in vitro assay established that the extract possessed potent activity with IC50 of 0.46 µg/mL compared to its DCM, butanol fractions and acarbose (IC50 5.3 µg/mL, 5.6 µg/mL, 45 µg/mL) respectively. BLAST analysis revealed the mushroom (accession number, KRO78278.1) to show 98% identity to Calvatia gigantea. The study established the identity of this mushroom and confirmed its antidiabetic activity.

Predicted potential occurrence of the North American false truffle Rhizopogon salebrosus in Europe

Abstract Rhizopogon salebrosus is an ectomycorrhizal fungus native to North America and known in Europe. In this study we illustrate the worldwide biogeography of R. salebrosus based on sporocarp and ectomycorrhiza records. MaxEnt modelling was used to assess the distribution of the potential niche of R. salebrosus in Europe, based on climatic variables and the preferences of its ectomycorrhizal partner. At the beginning, R. salebrosus was only recorded in the mountainous regions (Alps and Elbe Mountains). In 2014 this fungus was recorded for the first time in Northern Poland and this is the first record of R. salebrosus in a lowland area. In Europe the most suitable habitats are in the western part of the Iberian Peninsula, British Isles, western edge of the Scandinavian Peninsula, Alps, Balkan and Carpathian Mountains and the Anatolian Peninsula. Considering climatic conditions the most important factor limiting the occurrence of R. salebrosus (with a 31.4% contribution) is precipitation in the coldest quarter. On the other hand, areas where the ectomycorrhizal partners of R. salebrosus occur, should be also considered as potential regions for its expansion in Europe.

Spores of puffball fungus Lycoperdon pyriforme as a reference standard of stable monodisperse aerosol for calibration of optical instruments.

Advanced air quality control requires real-time monitoring of particulate matter size and concentration, which can only be done using optical instruments. However, such techniques need regular calibration with reference samples. In this study, we suggest that puffball fungus (Lycoperdon pyriforme) spores can be utilized as a reference standard having a monodisperse size distribution. We compare the Lycoperdon pyriforme spores with the other commonly used reference samples, such as Al2O3 powder and polystyrene latex (PSL) microspheres. Here we demonstrate that the puffball spores do not coagulate and, thus, maintain the same particle size in the aerosol state for at least 15 minutes, which is enough for instrument calibration. Moreover, the puffball mushrooms can be stored for several years and no agglomeration of the spores occurs. They are also much cheaper than other calibration samples and no additional devices are needed for aerosol generation since the fungal fruiting body acts as an atomizer itself. The aforementioned features make the fungal spores a highly promising substance for calibration and validation of particle size analyzers, which outperforms the existing, artificially produced particles for aerosol sampling. Furthermore, the L. pyriforme spores are convenient for basic research and development of new optical measurement techniques, taking into account their uniform particle size and absent coagulation in the aerosol.

Bioactivity-Guided Isolation of Anti-Inflammatory Constituents of the Rare Mushroom Calvatia nipponica in LPS-Stimulated RAW264.7 Macrophages.

Calvatia species, generally known as puffball mushrooms, are used both as sources of food and as traditional medicine. Among the Calvatia genus, Calvatia nipponica (Agaricaceae) is one of the rarest species. Using bioassay-guided fractionation based on anti-inflammatory effects, five alkaloids (1 - 5), two phenolics (6 and 7), and a fatty acid methyl ester (8) were isolated from the fruiting bodies of C. nipponica. Compound 8 was identified from C. nipponica for the first time, and all isolates (1 - 8) were tested for inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Compound 7 showed mild inhibition while compound 8 significantly inhibited NO production with an IC50 value of 27.50 ± 0.08 μm. The mechanism of NO inhibition of compound 7 was simulated by molecular docking analysis against nitric oxide synthase (iNOS), which revealed the interactions of 7 with the key amino acid residue and the heme in the active site. With the most potent inhibition against LPS-induced inflammation, compound 8 was further investigated with respect to its mechanism of action, and the activity was found to be mediated through the inhibition of iNOS and COX-2 expression.

Chemical constituents from the rare mushroom Calvatia nipponica inhibit the promotion of angiogenesis in HUVECs

Calvatia species, also known as puffball mushrooms, are common sources of food worldwide. Calvatia nipponica (Agaricaceae) is one of the most rare species in the Calvatia genus. It was first validated in 2008. Due to its scarcity, C. nipponica has never been chemically investigated. Its recent discovery in Korea allowed to conduct this study, and using bioactivity-guided fractionation for antiangiogenic activity, chemical investigation of the MeOH extract of the fruiting bodies of C. nipponica led to the isolation of five alkaloids (1–5) and two phenolic compounds (6–7). This is the first study to report the chemical investigation of C. nipponica, and compound 1 was previously reported as chemically synthesized only until our report of its isolation and identification from natural sources. The structure of 1 was determined by spectroscopic analysis by 1D and 2D NMR spectra and HR-MS. All compounds (1–7) were tested for inhibition of angiogenesis using human umbilical vein endothelial cells (HUVECs). Compounds 2, 4 and 5 significantly inhibited the promotion of angiogenesis in HUVECs. Compound 5 showed the most potent inhibition via downregulation of VEGF, p38 and ERK signaling pathways. These results suggested that the rare mushroom C. nipponica might be beneficial in antiangiogenesis therapy for cancer treatment.

Non-equilibrium cluster growth: where did it all begin

In this paper, Forrest and Witten reported on an experimental study of metal particle aggregates formed when a metal vapor produced by heating a plated filament was quench condensed. Metal particles of an average radius 40 A were streamed outward and accumulated in a thin spherical shell (a 'puff ball') of radius ~1 cm where the particles coalesced. The resulting particle aggregates moved down under gravity to an electron microscope slide where they were subsequently examined.

Restingomyces, a new sequestrate genus from the Brazilian Atlantic rainforest that is phylogenetically related to early-diverging taxa in Trappeaceae (Phallales)

Restingomyces reticulatus gen. et sp. nov. is a recently discovered false truffle species from Atlantic “restinga” rainforest in northeastern Brazil. Molecular and morphological characters separate this new sequestrate species from other described taxa in the order Phallales (Phallomycetidae, Basidiomycota). In our phylogenetic analysis based on nuc 28S rDNA and atp6, R. reticulatus forms a sister clade to Trappea darkeri and Phallobata alba, with the three taxa forming the earliest diverging lineage within Phallales. Morphological and molecular data warrant the recognition of the new genus and species, described here, and we also amend the taxonomic description for the family Trappeaceae.

New species of Elaphomyces (Elaphomycetaceae, Eurotiales, Ascomycota) from tropical rainforests of Cameroon and Guyana.

The sequestrate false truffles Elaphomyces favosus, E. iuppitercellus, and E. labyrinthinus spp. nov. are described as new to science from the Dja Biosphere Reserve, Cameroon. Elaphomyces adamizans sp. nov. is described as new from the Pakaraima Mountains of Guyana. The Cameroonian species are the first Elaphomyces taxa to be formally described from Africa, occurring in lowland Guineo-Congolian tropical rainforests dominated by the ectomycorrhizal (ECM) canopy tree Gilbertiodendron dewevrei (Fabaceae subfam. Caesalpinioideae). The Guyanese species is the third to be discovered in lowland tropical South America, occurring in forests dominated by the ECM trees Pakaraimaea dipterocarpacea (Dipterocarpaceae) and Dicymbe jenmanii (Fabaceae subfam. Caesalpinioideae). Macromorphological, micromorphological, habitat, and DNA sequence data are provided for each new species. Molecular and morphological data place these fungi in Elaphomycetaceae (Eurotiales, Ascomycota). Unique morphological features are congruent with molecular delimitation of each of the new species based on a phylogenetic analysis of the rDNA ITS and 28S loci across the Elaphomycetaceae. The phylogenetic analysis also suggests that a common ancestor is shared between some Elaphomyces species from Africa and South America, and that species of the stalked, volvate genus Pseudotulostoma may be nested in Elaphomyces.