Basidiomycete Genus Research Articles

An Economic and Reliable PCR Approach to Distinguish the Primary Pathogens Armillaria mellea and A. ostoyae From Other European Armillaria Species

ABSTRACTArmillaria is a soil‐borne genus of basidiomycetes whose species can cause stem and root rot in woody plants. The effects of plant‐pathogenic Armillaria species are well known in forests, but are underestimated in urban areas, where cases causing damage to trees and shrubs in green spaces have been steadily increasing in Switzerland since the 1980s. In this study, we present a simple, rapid, and cost‐effective protocol for high‐throughput diagnostics of the two primary pathogens A. mellea and A. ostoyae based on partial PCR amplification of the RPB2 gene. The specificity and sensitivity of the presented duplex PCR‐I and single‐plex PCR‐II were evaluated using different methods: (i) testing both PCRs on tree pathogenic or soil‐borne fungi of genera other than Armillaria, (ii) using dilution series of Armillaria‐DNA to determine a minimum detection limit, and (iii) sequencing the selected RPB2 region to verify the primer sequences and positions. The utility of PCR‐I and PCR‐II as a high‐throughput method was successfully tested on 65 DNA samples of Armillaria from Switzerland. Finally, an uninvolved person compared both classical methods, pairing test and sequencing, with PCR‐I and PCR‐II in a blind test. This study provides a reliable and alternative protocol for the rapid diagnosis of A. mellea and A. ostoyae causing root rot of woody plants.

Three new wood-inhabiting fungi of Botryobasidium (Cantharellales, Basidiomycota) from subtropical forests of Southwestern China.

The basidiomycete genus Botryobasidium is a resupinate saprotrophic with a global distribution range from coniferous to broad-leaved forest ecosystems. Though numerous species have been reported from Eurasia and North America, few have been described from China. In the current work, phylogenetic analyses of Botryobasidium in China were conducted based on the dataset of the internal transcribed spacer (ITS) regions and the large subunit (LSU) of nuclear ribosomal RNA gene. Maximum likelihood and Bayesian analyses were used to reconstruct the phylogenetic tree, and three new species, namely Botryobasidiumacanthosporum, B.leptocystidiatum and B.subovalibasidium, were described from subtropical forests of Yunnan Province, Southwestern China. Botryobasidiumacanthosporum is characterized by having yellowish white to dark yellow basidiome, clavate to tubular cystidia, and subglobose to globose basidiospores with obtuse spines. Botryobasidiumleptocystidiatum is characterized by having fluffy to arachnoid, greyish white to ivory basidiome, generative hyphae with clamped, tubular cystidia, and subnavicular to navicular basidiospores. While, B.subovalibasidium is characterized by having yellowish to ivory basidiome, subovoid basidia, navicular to suburniform basidiospores, and thick-walled chlamydospores. These three new species are described and illustrated, and the discriminating characters between the new species and their closely related species are discussed. A key to known species of Botryobasidium in China is provided.

The diversity and taxonomy of Tomentella (Thelephoraceae, Thelephorales) with descriptions of four new species from Southwestern China.

Taxonomy plays a central role in understanding the diversity of life, translating the products of biological exploration and discovery specimens and observations into systems of names that settle a "classification home" to taxa. Up to this point, studies on the taxonomy and phylogeny of the basidiomycetous genus Tomentella stemmed mainly from the temperate to boreal zones of the North Hemisphere, but were scarce in tropical Asia. In this study, four new species, viz. Tomentellaolivaceobasidiosa, T.velutina, T.wumenshanensis and T.yunnanensis from China, are described and illustrated based on the morphological characteristics and molecular phylogenetic analyses, in which the sequences of ITS+nLSU+mtSSU+RPB2 genes were used for the phylogenetic analyses using Maximum Likelihood, Maximum Parsimony and Bayesian Inference methods. All the new species can be well recognised by their macroscopical and anatomical characteristics. The four new species, closely related taxa in the phylogenetic tree and morphologically similar species are discussed.

Four New Species of Tomentella (Thelephorales, Basidiomycota) from Subtropical Forests in Southwestern China.

Species of the basidiomycetous genus Tomentella are widely distributed throughout temperate forests. Numerous studies on the taxonomy and phylogeny of Tomentella have been conducted from the temperate zone in the Northern hemisphere, but few have been from subtropical forests. In this study, four new species, T. casiae, T. guiyangensis, T. olivaceomarginata and T. rotundata from the subtropical mixed forests of Southwestern China, are described and illustrated based on morphological characteristics and phylogenetic analyses of the internal transcribed spacer regions (ITS) and the large subunit of the nuclear ribosomal RNA gene (LSU). Molecular analyses using Maximum Likelihood and Bayesian analysis confirmed the phylogenetic positions of these four new species. Anatomical comparisons among the closely related species in phylogenetic and morphological features are discussed. Four new species could be distinguished by the characteristics of basidiocarps, the color of the hymenophoral surface, the size of the basidia, the shape of the basidiospores and some other features.

Decoding the RNA virome of the tree parasite Armillaria provides new insights into the viral community of soil-borne fungi.

The globally distributed basidiomycete genus Armillaria includes wood decomposers that can act as opportunistic parasites, causing deadly root rot on woody plants. To test whether RNA viruses are involved in this opportunistic behaviour, a large isolate collection of five Armillaria species collected over 40 years in Switzerland from trees, dead wood and soil was analysed. De novo assembly of RNA-Seq data revealed 21 viruses, 14 of which belong to putative new species. Two dsRNA viruses and an unclassified Tymovirales are formally described for the first time for Armillaria. One mitovirus occurred with a high prevalence of 71.1%, while all other viruses were much less prevalent (0.6%-16.9%). About half of all viruses were found only in one fungal species, others occurred in 2-6 fungal species. Co-infections of 2-7 viruses per isolate were not uncommon (34.9%), and most viruses persisted circulating within fungal populations for decades. Some viruses were related to viruses associated with other Armillaria species, supporting the hypothesis that virus transmission can occur between different fungal species. Although no specific correlation between viruses and the fungal trophic strategy was found, this study opens new insights into viral diversity hidden in the soil microbiome.

Revision of Kneiffiella with segregation of Egonia gen. nov. (Hymenochaetales, Agaricomycetes): how similar morphology can hide taxonomic diversity in the molecular era

DNA-based phylogenetic inference has shown that many morphological traits, once considered taxonomically relevant, are the result of convergent evolution. In this study, we analyze Kneiffiella, a genus of corticioid and polyporoid wood-inhabiting basidiomycetes in the order Hymenochaetales. A multi-gene-based phylogeny demonstrates that Kneiffiella is polyphyletic, and the new genus Egonia is established for K. cineracea and its relatives, despite their morphological similarity to other Kneiffiella species. Additionally, we present the best resolved phylogram of the Hymenochaetales to date. A revised circumscription of Kneiffiella is presented, as well as an overview of the 29 species accepted in the genus, including 10 new species. Three new species of Egonia are described, and H. subefibulata is placed in synonymy with K. subglobosa. A key to the species of Kneiffiella and Egonia is presented.

Characterization of fungal species involved in white haze disorder on apples in Northern Italy and description of Golubevia mali sp. nov. and Entyloma mali sp. nov.

White haze, caused by an extensive fungal colonization of the apple surface, is an emerging postharvest issue in several European apple production areas. It results in compromised quality and decreased marketability of the fruits, leading to economic losses. In this study, the occurrence and the diversity of white haze-related fungi associated with apples was investigated in Northern Italy. Fungal strains were isolated from apple tissues and the species diversity was assessed using molecular and phylogenetic tools. Moreover, the ability of the isolated species to reproduce symptoms on healthy apples artificially inoculated was tested. Seventy-two fungal isolates were collected. Six species belonging to different basidiomycetous genera (Entyloma, Golubevia, Tilletiopsis) were identified, showing high diversity of fungi involved in white haze development in Northern Italy. The strains were identified as E. belangeri, E. randwijkense, G. pallescens, T. washingtonensis. Moreover, two new species, E. mali sp. nov. and G. mali sp. nov. were described. The most frequently isolated strains were inoculated on healthy apple fruit, showing to be able to reproduce symptoms on red-skin apples, fulfilling Koch’s postulates. This work provides new insights to increase knowledge about the causal agents of white haze on apple. In addition, the names Golubevia, Golubeviaceae and Golubeviales have been nomenclaturally validated.

Frequent transitions in mating-type locus chromosomal organization in Malassezia and early steps in sexual reproduction

Fungi in the basidiomycete genus Malassezia are the most prevalent eukaryotic microbes resident on the skin of human and other warm-blooded animals and have been implicated in skin diseases and systemic disorders. Analysis of Malassezia genomes revealed that key adaptations to the skin microenvironment have a direct genomic basis, and the identification of mating/meiotic genes suggests a capacity to reproduce sexually, even though no sexual cycle has yet been observed. In contrast to other bipolar or tetrapolar basidiomycetes that have either two linked mating-type-determining (MAT) loci or two MAT loci on separate chromosomes, in Malassezia species studied thus far the two MAT loci are arranged in a pseudobipolar configuration (linked on the same chromosome but capable of recombining). By generating additional chromosome-level genome assemblies, and an improved Malassezia phylogeny, we infer that the pseudobipolar arrangement was the ancestral state of this group and revealed six independent transitions to tetrapolarity, seemingly driven by centromere fission or translocations in centromere-flanking regions. Additionally, in an approach to uncover a sexual cycle, Malassezia furfur strains were engineered to express different MAT alleles in the same cell. The resulting strains produce hyphae reminiscent of early steps in sexual development and display upregulation of genes associated with sexual development as well as others encoding lipases and a protease potentially relevant for pathogenesis of the fungus. Our study reveals a previously unseen genomic relocation of mating-type loci in fungi and provides insight toward the identification of a sexual cycle in Malassezia, with possible implications for pathogenicity.

Fungal succession in decomposing ash leaves colonized by the ash dieback pathogen Hymenoscyphus fraxineus or its harmless relative Hymenoscyphus albidus.

The ascomycete Hymenoscyphus fraxineus, originating from Asia, is currently threatening common ash (Fraxinus excelsior) in Europe, massive ascospore production from the saprotrophic phase being a key determinant of its invasiveness. To consider whether fungal diversity and succession in decomposing leaf litter are affected by this invader, we used ITS-1 metabarcoding to profile changes in fungal community composition during overwintering. The subjected ash leaf petioles, collected from a diseased forest and a healthy ash stand hosting the harmless ash endophyte Hymenoscyphus albidus, were incubated in the forest floor of the diseased stand between October 2017 and June 2018 and harvested at 2-3-month intervals. Total fungal DNA level showed a 3-fold increase during overwintering as estimated by FungiQuant qPCR. Petioles from the healthy site showed pronounced changes during overwintering; ascomycetes of the class Dothideomycetes were predominant after leaf shed, but the basidiomycete genus Mycena (class Agaricomycetes) became predominant by April, whereas H. albidus showed low prevalence. Petioles from the diseased site showed little change during overwintering; H. fraxineus was predominant, while Mycena spp. showed increased read proportion by June. The low species richness and evenness in petioles from the diseased site in comparison to petioles from the healthy site were obviously related to tremendous infection pressure of H. fraxineus in diseased forests. Changes in leaf litter quality, owing to accumulation of host defense phenolics in the pathogen challenged leaves, and strong saprophytic competence of H. fraxineus are other factors that probably influence fungal succession. For additional comparison, we examined fungal community structure in petioles collected in the healthy stand in August 2013 and showing H. albidus ascomata. This species was similarly predominant in these petioles as H. fraxineus was in petioles from the diseased site, suggesting that both fungi have similar suppressive effects on fungal richness in petiole/rachis segments they have secured for completion of their life cycle. However, the ability of H. fraxineus to secure the entire leaf nerve system in diseased forests, in opposite to H. albidus, impacts the general diversity and successional trajectory of fungi in decomposing ash petioles.

New Inonotus Polysaccharides: Characterization and Anticomplementary Activity of Inonotus rheades Mycelium Polymers.

Inonotus is a small genus of xylotrophic basidiomycetes and a source of bioactive fungochemicals among which a special place is occupied by polymeric compounds. In this study, polysaccharides that are widespread in Europe, Asia, and North America and a poorly understood fungal species, I. rheades (Pers.) Karst. (fox polypore), were investigated. Water-soluble polysaccharides of I. rheades mycelium were extracted, purified, and studied using chemical reactions, elemental and monosaccharide analysis, UV-Vis and FTIR spectroscopy, gel permeation chromatography, and linkage analysis. Five homogenic polymers (IRP-1-IRP-5) with molecular weights of 110-1520 kDa were heteropolysaccharides that consist mainly of galactose, glucose, and mannose. The dominant component, IRP-4, was preliminary concluded to be a branched (1→3,6)-linked galactan. Polysaccharides of I. rheades inhibited the hemolysis of sensitized sheep erythrocytes by complement from human serum, signifying anticomplementary activity with the greatest effects for the IRP-4 polymer. These findings suggest that I. rheades mycelium is a new source of fungal polysaccharides with potential immunomodulatory and anti-inflammatory properties.

The bHLH-zip transcription factor SREBP regulates triterpenoid and lipid metabolisms in the medicinal fungus Ganoderma lingzhi

Ganoderic acids (GAs) are well recognized as important pharmacological components of the medicinal species belonging to the basidiomycete genus Ganoderma. However, transcription factors directly regulating the expression of GA biosynthesis genes remain poorly understood. Here, the genome of Ganoderma lingzhi is de novo sequenced. Using DNA affinity purification sequencing, we identify putative targets of the transcription factor sterol regulatory element-binding protein (SREBP), including the genes of triterpenoid synthesis and lipid metabolism. Interactions between SREBP and the targets are verified by electrophoretic mobility gel shift assay. RNA-seq shows that SREBP targets, mevalonate kinase and 3-hydroxy-3-methylglutaryl coenzyme A synthetase in mevalonate pathway, sterol isomerase and lanosterol 14-demethylase in ergosterol biosynthesis, are significantly upregulated in the SREBP overexpression (OE::SREBP) strain. In addition, 3 targets involved in glycerophospholipid/glycerolipid metabolism are upregulated. Then, the contents of mevalonic acid, lanosterol, ergosterol and 13 different GAs as well as a variety of lipids are significantly increased in this strain. Furthermore, the effects of SREBP overexpression on triterpenoid and lipid metabolisms are recovered when OE::SREBP strain are treated with exogenous fatostatin, a specific inhibitor of SREBP. Taken together, our genome-wide study clarify the role of SREBP in triterpenoid and lipid metabolisms of G. lingzhi.

Fungal outbreak in the Catacombs of SS. Marcellino and Pietro Rome (Italy): From diagnosis to an emergency treatment.

The present study reports a sudden fungal outbreak that occurred in the corridor near the entrance of the Catacombs of SS. Marcellino and Pietro in Rome (Italy) observed after 1 year of a restoration treatment that interested the walls of the entrance of the Catacombs and some artifacts placed in situ. The colonization was observed on the vault at the entrance and in correspondence with the restored marble pieces displayed on the left side of the corridor. No growth was observed on the right side where similarly treated marble slabs were placed. Samples taken in correspondence with fungal biofilm were analyzed through the combined use of microscopical, cultural, and molecular tools and showed that the vault and the left side of the corridor entrance were colonized by a complex fungal biofilm consisting mainly of Coniophora sp. and other genera, such as Hypomyces, Purpureocillium, Acremonium, Penicillium, and Alternaria, many of which are well known as responsible of biodeterioration of stone surfaces. Regarding the brown-rot basidiomycete Coniophora, it was able to form very large colonies on the substrata with a diameter of up to 57 cm. Although the direct observation under a light microscope evidenced the presence of abundant brown fungal conidia, several attempts to cultivate the microorganism failed, therefore only through DNA sequencing analyses, it was possible to identify and characterize this fungus. There is very little literature on the genus Coniophora which is reported as one of the causes of wet-rot decay of wood in buildings. A connection with calcium-containing materials such as bricks and mortars was demonstrated, but no data were available about the possible role of this species in the biodeterioration of stones. This study features the first finding of a strain related to the basidiomycetous genus of Coniophora in the order Boletales in association with evident phenomena of biodeterioration.

Photochemical defense as trait of fungi from Cortinarius subgenus Dermocybe

The photobiological activity of ten colorful species belonging to subgenus Dermocybe of the basidiomycete genus Cortinarius was investigated. Extracts of all species produced singlet oxygen and are thus photoactive. Pigment analysis was performed and showed similarities of the anthraquinone pigments across the species in dependency to their respective pigmentation types. Detailed content analysis of the pigments in the whole agaricoid fruiting body compared to the three different tissue types (pileus, stipe, and lamellae) revealed that the pigments emodin, dermocybin, and dermorubin, as well as their respective glycosides, are enhanced in the gills. In an independent experiment, the gills were shown to be the most photoactive tissues of the fruiting body. Photobiological experiments with invertebrates (i.e., glassworm Chaoborus crystallinus) proved a phototoxic effect of the methanolic extract of the red blood webcap (Cortinarius sanguineus var. aurantiovaginatus). This work adds further evidence to a common photobiological trait in Cortinarius subgenus Dermocybe and underpins the possibility of a photochemical defense mechanism in fungi.Graphical abstract

Immunomodulatory Effect of Polysaccharides from the Mushroom-Forming Basidiomycete Gymnopilus imperialis (Agaricomycetes, Basidiomycota).

Gymnopilus consists of a widely distributed genus of basidiomycetes, especially in tropical regions of the world, such as Japan, Australia, Paraguay, and Brazil. This genus biosynthesizes interesting bioactive compounds, such as sesquiterpenoids, oligoisoprenoids, styrylpyrones, and lectins. In the present study, the aqueous extract of the basidiomata of Gymnopilus imperialis (Basidiomycota, Agaricomycetes, Agaricales, Hymenogastraceae) was obtained by using the accelerated solvent extraction (ASE) technique, followed by the precipitation of polysaccharide fraction with ethanol. Further purification by freeze-thawing processes, Fehling solution precipitation, and membrane dialysis with different pore sizes yield three main polysaccharide fractions (Gi-MRSW, Gi-PFME, and Gi-SFME). According to monosaccharide composition and 13C-NMR data, the Gi-MRSW and Gi-SFME fractions showed to be composed mainly of β-glucans and Gi-PFME by a heterogalactan. Moreover, the immunomodulatory potential of Gi-MRSW was evaluated using RAW 264.7 murine macrophage as a study model. The nitric oxide production was significantly increased in treated samples, and the expression of inducible nitric oxide synthase (iNOS) showed that the fraction Gi-MRSW from G. imperialis induces the M1 polarization phenotype.

Six New Species of Tomentella (Thelephorales, Basidiomycota) From Tropical Pine Forests in Central Vietnam.

Up to this point, studies on the taxonomy and phylogeny of the basidiomycetous genus Tomentella stemmed mainly from the temperate to boreal zones of the Northern hemisphere but were scarce in tropical Asia. In this study, six new species—T. bidoupensis, T. brevisterigmata, T. cinereobrunnea, T. longiechinula, T. stipitobasidia, and T. verruculata from central Vietnam in Southeast Asia—are described and illustrated on the basis of morphological characteristics and molecular phylogenetic analyses of the nuclear ribosomal ITS (internal transcribed spacer: ITS1-5.8S-ITS2) and LSU (large subunit: 28S) markers. Maximum likelihood and Bayesian analyses were used to confirm the phylogenetic positions of these new species and all of them can be well recognized by the macroscopical and anatomical characteristics. The new species and closely related species in the phylogenetic tree, and the new species and morphologically similar species are discussed, whereas the host plant for these new species were speculated on the basis of the phylogenetic analyses and the tree species information of the investigated forests.

The complete genome sequence of a novel mycovirus in the plant-pathogenic fungus Exobasidium gracile.

The plant-pathogenic fungus Exobasidium gracile belongs to the basidiomycetous genus Exobasidium and causes leaf abnormalities in Camellia oleifera. Here, we report a novel mycovirus from E. gracile strain Z-2, called "Exobasidium gracile zybavirus 2" (EgZV2), which is related phylogenetically to a member of the genus Zybavirus in the family Amalgaviridae. Like those of other zybaviruses, the EgZV2 genome has two large open reading frames (ORFs) encoding an unknown protein and an RNA-dependent RNA polymerase (RdRp), respectively. In addition, we found a conserved +1 programmed ribosomal frameshifting (PRF) motif (UUUCGG) between ORF1 and ORF2 in EgZV2.

Molecular characterization of three novel mycoviruses in the plant pathogenic fungus Exobasidium

The plant pathogen Exobasidium gracile, which belongs to the basidiomycetous genus Exobasidium, can lead to swollen and thicker leaves of C. oleifera. To our knowledge, there have been no reports of mycoviruses infecting Exobasidium gracile. This study characterized three mycoviruses coinfecting the plant pathogen Exobasidium gracile strain Z-1. Based on phylogenetic and genomic analyses, E. gracile strain Z-1 was infected two putative Totiviruses designated Exobasidium gracile Totivirus 1 (EgTV1) and Exobasidium gracile Totivirus 2 (EgTV2) and a putative Zybavirus of the family Amalgaviridae defined Exobasidium gracile Zybavirus 1 (EgZV1). Similar to the genomic organization of other Totiviruses, the EgTV1 and EgTV2 genomes are composed of one dsRNA segment that exhibits two large ORFs encoding a CP (capsid protein) and an RdRp (RNA-dependent RNA polymerase), respectively. Moreover, EgTV1 and EgTV2 genomes with a candidate -1 slippery heptamer sequence were discovered between CP and RdRp, respectively. Similar to other Zybaviruses of the family Amalgaviridae, the EgZV1 genome is composed of one dsRNA segment that contains two large ORFs encoding an unknown protein and an RdRp. In addition, the EgZV1 genome has a candidate +1 slippery heptamer sequence between an unknown protein and RdRp.

Phaffia brasiliana sp. nov., a yeast species isolated from soil in a Cerrado-Atlantic Rain Forest ecotone site in Brazil.

During studies of yeasts associated with soil in a Cerrado-Atlantic Rain Forest ecotone site in Brazil, three orange-pigmented yeast strains were isolated from samples collected in Minas Gerais state, Brazil. Molecular analyses combining the 26S rRNA gene (D1/D2 domains) and the internal transcribed spacer (ITS) sequences as well as whole-genome sequence data showed that these strains could not be ascribed to any known species in the basidiomycetous genus Phaffia, and thus they are considered to represent a novel species for which the name Phaffia brasiliana sp. nov. is proposed. The holotype is CBS 16121T and the MycoBank number is MB 839315. The occurrence of P. brasiliana in a tropical region is unique for the genus, since all other species occur in temperate regions. Two factors appear to contribute to the distribution of the novel taxon: first, the region where it was found has relatively moderate temperature ranges and, second, an adaptation to grow or withstand temperatures higher than those of the other species in the genus seems to be in place.

Dereplication of extracts from nutraceutical mushrooms Pleurotus using molecular network approach

Pleurotus is an edible mushroom from the well-known genus of Basidiomycetes; it is the second-most commonly consumed mushroom worldwide. This genus is characterized by the presence of steroids, fatty acids, and polysaccharides. Recently, Pleurotus has become popular as a functional food owing to its health benefits, primarily because they are a source of vitamins, fibers, minerals, and lipids. In natural products chemistry, dereplication techniques identify bioactive molecules from natural sources such as plants and fungi without isolating or characterizing molecules. We used dereplication techniques aided by the Global Natural Products Social Molecular Network to analyze the chemical composition of the methanolic extracts of six Pleurotus species (P. sapidus, P. ostreaus, P. ostreaus var. Florida, P. djamor, P. citrinopileatus, and P. pulmonarius), to identify bioactive molecules with nutraceutical properties. Using this technique, we identified several molecular families, including eight fatty acids and seven steroids. Our findings suggest that dereplication is a relatively rapid tool for characterizing fungal species and determining their nutraceutical value.

Cryptococcus depauperatus, a close relative of the human-pathogen C. neoformans, associated with coffee leaf rust (Hemileia vastatrix) in Cameroon.

The genus Cryptococcus is well known for its two species -Cryptococcus neoformans and C. gatii-that are etiological agents of cryptococcosis,an important fungal disease of mammals, including humans, and which is particularly common in immunocompromised patients. Nevertheless, Cryptococcus is a large and widely distributed genus of basidiomycetes occupying a broad range of niches, including mycoparasitism. One such mycoparasitic species is Cryptococcus depauperatus, which was firstly mistakenly described as a pathogen of scale insects under the name Aspergillus depauperatus. The "Aspergillus" conidiophores were later shown to be basidia of a Cryptococcus and the new combination C. depauperatus was proposed. Additionally, instead of an entomopathogen, the fungus was found to be a mycoparasite growing on the entomopathogen Akanthomyces (Lecanicillium) lecanii. Recently, during surveys for mycoparasites of coffee leaf rust (Hemileia vastatrix) in the context of a biocontrol project, white colonies covering rust pustules were observed in Cameroon. Upon close examination, instead of a member of the "white colony forming complex" of Ascomycetes, commonly collected growing on H. vastatrix, such colonies were found to represent a basidiomycete fungus with basidia-bearing chains of basidiospores, typical of the genus Cryptococcus. Morphological and molecular evidence was generated supporting the identification of the fungus on rust pustules as C. depauperatus. This is the first record of C. depauperatus from Africa and of its association with coffee leaf rust.