Isolation, identification and bioactivity analysis of an endophytic fungus isolated from Aloe vera collected from Asir desert, Saudi Arabia.

Endophytic fungi isolated from desert plants are among the less known organisms with potentially valuable applications. The bioactivities of an endophytic fungus isolated from Aloe vera, a plant found in central regions of Asir desert, Saudi Arabia. Based on primary phytochemical screening, an efficient isolate was selected and identified according to the sequence analysis of the internal spacer regions ITS1, ITS4 and the 5.8S region as Preussia africana belonging to the family Sporormiaceae. The crude extract of this fungus was evaluated for its bioactivities. Under static conditions, the crude extract at a concentration of 500 µg/mL had a strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging rate of 87%, whereas a higher concentration (100 µg/mL) had an astounding wound healing effect (42.6% at 48 h) when compared to positive control. Moreover, the crude extract with a concentration of 50 µg/mL was active against almost all cancer cell lines such as HeLa (cervical cancer), Hep G2 (liver cancer), MCF-7 (breast cancer), A549 (lung cancer), LN-229 (glioblastoma), A-431 (skin cancer), and kidney cell line (HEK 293T). The results suggest that the endophytic fungus P. africana from A. vera has wide therapeutic applications against severe disease conditions.

Remotididymella ageratinae sp. nov. and Remotididymella anemophila sp. nov., two novel species isolated from the invasive weed Ageratina adenophora in PR China.

To determine if Ageratina adenophora can accumulate diverse pathogens from surrounding native plants, we intensively sampled fungal communities, including endophytes, leaf spot pathogens and canopy air fungi, associated with Ag. adenophora as well as native plants in its invasive range. In total, we collected 4542 foliar fungal strains from 10 geographic sites, including 1340 from healthy leaves of Ag. adenophora, 2051 from leaf spots of Ag. adenophora and 1151 from leaf spots of 56 species of native plants and crops. Taxonomically, the common fungal genera included Colletotrichum, Diaporthe, Alternaria, Nemania, Xylaria, Neofusicoccum, Nigrospora, Epicoccum, Gibberella, Pestalotiopsis, Irpex, Schizophyllum and Clonostachys. We also isolated the cultivable fungi from 12 air samples collected from six areas in Yunnan Province, PR China. Among the total of 1255 air fungal isolates, the most common genera were Cladosporium, Trichoderma and Epicoccum. Among them, two new Remotididymella species, Remotididymella ageratinae from leaf spot of Ag. adenophora and Remotididymella anemophila from canopy air of Ag. adenophora were found. The two species showed both asexual and sexual reproductive structures. The conidia of R. ageratinae and R. anemophila are larger than those of R. anthropophila and R. destructiva. The size of ascospores of R. ageratinae and R. anemophila also differ from R. bauhiniae. Phylogenetic analysis of the combined ITS, LSU rRNA, rpb2 and tub2 sequences showed that R. ageratinae and R. anemophila each formed a distinct clade, separated from all species previously described in Remotididymella and confirmed them as new species belonging to Remotididymella. Full descriptions of R. ageratinae and R. anemophila are provided in this study.

Hepatic phaeohyphomycosis due to a novel dematiaceous fungus, Pleurostoma hongkongense sp. nov., and importance of antifungal susceptibility testing.

Pleurostoma species are wood-inhabiting fungi and emerging opportunistic pathogens causing phaeohyphomycosis. In this study, we isolated a dematiaceous fungus, HKU44T, from the subhepatic abscess pus and drain fluids of a liver transplant recipient with post-transplant biliary and hepatico-jejunostomy bypass strictures. Histology of the abscess wall biopsy showed abundant fungal hyphae. The patient survived after a second liver transplant and antifungal therapy. On SDA, HKU44T grew initially as white powdery colonies which turned beige upon maturation. Hyphae were septate and hyaline. Phialides were monophialidic and laterally located, generally closely associated to a cluster of conidia which were usually reniform. Phylogenetic analyses showed that HKU44T is most closely related to, but distinct from, Pleurostoma ootheca and Pleurostoma repens. These suggested that HKU44T is a novel Pleurostoma species, for which the name Pleurostoma hongkongense sp. nov. is proposed. Antifungal susceptibility testing showed that Pleurostoma species possessed high MICs/MECs for fluconazole, 5-flucytosine and the echinocandins; whereas they exhibited a high strain-to-strain variability to the susceptibilities to the other triazoles. As for amphotericin B, ∼65% of the Pleurostoma strains had low MICs (≤1 µg/mL). DNA sequencing should be performed to accurately identify fungi with Pleurostoma/Phialophora-like morphologies, so is antifungal susceptibility testing for patients with Pleurostoma infections.

Neoscytalidium dimidiatum un raro agente de onicomicosis no dermatofítica en Chile / Neoscytalidium dimidiatuma rare agent ofnon dermatophyticonychomycosisin Chile

El término de onicomicosis se emplea para describir las infecciones de las uñas causadas por diferentes grupos taxonómicos fúngicos ya sea filamentosos como levaduriformes. A pesar de que estas patologías son causadas en los vertebrados principalmente por integrantes de la Familia Artrodermatáceae (Onygenales), la micología médica aplicó para ellos la terminología más específica de dermatofitosis, por ser un grupo ecológico de mayor importancia y presencia clínica. Las dermatomicosis de piel y fanéreos, representan un conjunto de infecciones producidas por especies fúngicas distribuidas en ambientes diversos, capaces de crecer a temperaturas de 37° y que actúan usualmente como patógenos oportunistas cuando existe generalmente un factor predisponente en el huésped. Se destaca la colonización en una uña de los pies en un hombre de 49 años por Neoscytalidium dimidiatum(Penz.) Crous & Slippers, un reconocido fitopatógeno de rápido crecimiento, común en zonas tropicales y subtropicales, que presentó la capacidad de invadir tejidos queratinizados con un aspecto clínico indistinguible de los causadas por dermatofitos. Por la rara presencia de este hongo en nuestra zona geográfica (provincia de Valparaíso, Chile), se aportan los principales datos morfofisiológicos,taxonómicos y moleculares utilizados en su diagnóstico. (AU)

The term onychomycosis is used to describe nail infections caused by different fungal taxonomic groups, either filamentous or yeast. Despite the fact that these pathologies are caused in vertebrates mainly by members of the Artrodermatáceae Family (Onygenales), medical mycology applied the more specific terminology of dermatophytosis for them, as it is an ecological group of greater importance and clinical presence. Skin and pharynx dermatomycosis represent a set of infections produced by fungal species distributed in diverse environments, capable of growing at temperatures of 37° and that usually act as opportunistic pathogens when there is a predisposing factor in the host. The colonization on a toenail in a 49-year-old man by Nesoscytalidium dimidiatumis highlighted (Penz.) Crous & Slippers, a recognized fast-growing phytopathogen, common in tropical and subtropical areas, which presented the ability to invade keratinized tissues with a clinical appearance indistinguishable from those caused by dermatophytes. Due to the rare presence of this fungus in our geographical area (Valparaíso province, Chile), the main morphophysiological, taxonomic and molecular data used in its identificationare provided. (AU)

Identification and Functional Characterization of a Novel Immunomodulatory Protein From Morchella conica SH.

A novel fungal immunomodulatory protein (FIP) was found in the precious medical and edible mushroom Morchella conica SH, defined as FIP-mco, which belongs to the FIP family. Phylogenetic analyses of FIPs from different origins were performed using Neighbor-Joining method. It was found that FIP-mco belonged to a new branch of the FIP family and may evolved from a different ancestor compared with most other FIPs. The cDNA sequence of FIP-mco was cloned and expressed in the yeast Pichia Pastoris X33. The recombinant protein of FIP-mco (rFIP-mco) was purified by agarose Ni chromatography and determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. The protein rFIP-mco could significantly suppress the proliferation of A549 and HepG2 cells at the concentration of 15 and 5 µg/ml, respectively, and inhibited the migration and invasion of human A549 and HepG2 cells at the concentration of 15 and 30 µg/ml respectively in vitro. Further, rFIP-mco can significantly reduce the expression levels of TNF-α, IL-1ß, and IL-6 in the THP1 cells (human myeloid leukemia mononuclear cells). In order to explore the potential mechanism of the cytotoxicity effect of rFIP-mco on A549 and HepG2 cells, cell cycle and apoptosis assay in the two cancer cells were conducted. The results demonstrated that G0/G1 to S-phase arrest and increased apoptosis may contribute to the proliferation inhibition by rFIP-mco in the two cancer cells. Molecular mechanism of rFIP-mco's reduction effect on the inflammatory cytokines was also studied by suppression of the NF-κB signaling pathway. It showed that suppression of NF-κB signaling is responsible for the reduction of inflammatory cytokines by rFIP-mco. The results indicated the prospect of FIP-mco from M. conica SH as an effective and feasible source for cancer therapeutic studies and medical applications.

The new truffle genus Babosia and a new species of Stouffera from semiarid grasslands of Hungary.

Truffles with distinct morphological and anatomical features were collected during a study of hypogeous fungi of semiarid sandy grasslands of the Great Hungarian Plain in Hungary, representing the westernmost localities of the Eurasian steppe belt. None of the ascomata were collected near ectomycorrhizal plant species, and none were identified as ectomycorrhizal during previous surveys in the collection area. We studied morphoanatomical characteristics of these truffles with light and scanning electron microscopy and investigated their phylogenetic positions based on analyses of different nuclear loci. The truffles were found to represent two novel lineages that grouped with the Marcelleina-Peziza gerardii clade of the Pezizaceae. One formed a distinct lineage, for which we propose a new genus Babosia with a new species Babosia variospora characterized by diverse spore ornamentation varying even within one ascus. The truffles in the other lineage clustered with the rarely collected American truffle Stouffera longii and share with it similar spore ornamentation and habitat features. However, our material differs from S. longii by geographic origin, the quick and strong coloration of the ascomata to dark gray at cut surface or bruised area, varying spore number in asci, and smaller spore size; thus, we describe it as a new species, Stouffera gilkeyae.

A new species of Gloeandromyces from Ecuador and Panama revealed by morphology and phylogenetic reconstruction, with a discussion of secondary barcodes in Laboulbeniomycetes taxonomy.

This paper describes and illustrates a new species of Laboulbeniales (Ascomycota, Laboulbeniomycetes) recovered from Mastoptera guimaraesi bat flies (Diptera, Streblidae) in Ecuador and Panama. Bat fly-associated Laboulbeniales are still unexplored in the Neotropics, with only four described species of Gloeandromyces and one species of Nycteromyces known. Morphological characteristics and phylogenetic analyses support placement of the new taxon in Gloeandromyces and its recognition as an undescribed species. Gloeandromyces hilleri sp. nov. is easily recognized by 2-3 longitudinal rows of undulations at its perithecial venter. Phylogenetic reconstructions of the large subunit (LSU) ribosomal DNA and the translation elongation factor 1α (TEF1) both resolve G. hilleri and G. nycteribiidarum as sister species. We discuss the utility of LSU and TEF1 as secondary barcodes in Laboulbeniomycetes taxonomy.

Species diversity of Pleosporalean taxa associated with Camellia sinensis (L.) Kuntze in Taiwan.

Pleosporales species are important plant pathogens, saprobes, and endophytes on a wide range of economically important plant hosts. The classification of Pleosporales has undergone various modifications in recent years due to the addition of many families described from multiple habitats with a high level of morphological deviation. Numerous asexual genera have been described in Pleosporales that can be either hyphomyceteous or coelomycetous. Phoma- or coniothyrium-like species are common and have been revealed as polyphyletic in the order Pleosporales and linked with several sexual genera. A total of 31 pleosporalean strains were isolated in different regions of Taiwan between 2017 and 2018 from the leaves of Camellia sinensis plants with symptoms of leaf spot disease. These strains were evaluated morphologically and genotypically using multi-locus sequence analyses of the ITS, LSU, SSU, rpb2, tef1 and tub2 genes. The results demonstrated the affiliation of these strains with the various families in Pleosporales and revealed the presence of one new genus (Neoshiraia) and eight new species (Alloconiothyrium camelliae, Amorocoelophoma camelliae, Leucaenicola camelliae, L. taiwanensis, Neoshiraia camelliae, N. taiwanensis, Paraconiothyrium camelliae and Paraphaeosphaeria camelliae). Furthermore, to the best of our understanding, Didymella segeticola, Ectophoma pomi and Roussoella mexican were reported for the first time from C. sinensis in Taiwan.

Medicopsis romeroi nodular subcutaneous infection in a kidney transplant recipient.

Phaeohyphomycosis is a set of fungal infections caused by various dematiaceous fungi such as coelomycetes. These infections can occur either in immunocompetent or immunocompromised patients like solid organ transplants. Here we describe a nodular lesion of the right hallux that occurred in a kidney transplant patient. Microscopic examination of the biopsy revealed fungal hyphae and culture was positive to a grey to black mould that lacked characteristic elements to be identified. Nucleic acid sequencing targeting the internal transcribed spacer of the ribosomal DNA identified this mould as Medicopsis romeroi. The patient benefited of an antifungal therapy with voriconazole associated with surgical excision of the lesion. No relapse of the lesion was observed during a six-month follow-up. In solid organ transplants, phaeohyphomycosis caused by Medicopsis romeroi are very rare with only 12 cases reported. The clinical history should be well assessed since the lesion can appear several years after a cutaneous trauma that happened in a tropical region. Therapy generally combines antifungals with surgical excision of the lesion in order to avoid any relapse or dissemination of the infection.

Characterization of Nuclear and Mitochondrial Genomes of Two Tobacco Endophytic Fungi Leptosphaerulina chartarum and Curvularia trifolii and Their Contributions to Phylogenetic Implications in the Pleosporales.

The symbiont endophytic fungi in tobacco are highly diverse and difficult to classify. Here, we sequenced the genomes of Curvularia trifolii and Leptosphaerulina chartarum isolated from tobacco plants. Finally, 41.68 Mb and 37.95 Mb nuclear genomes were sequenced for C. trifolii and L. chartarum with the scaffold N50, accounting for 638.94 Kb and 284.12 Kb, respectively. Meanwhile, we obtained 68,926 bp and 59,100 bp for their mitochondrial genomes. To more accurately classify C. trifolii and L. chartarum, we extracted seven nuclear genes and 12 mitochondrial genes from these two genomes and their closely related species. The genes were then used for calculation of evolutionary rates and for phylogenetic analysis. Results showed that it was difficult to achieve consistent results using a single gene due to their different evolutionary rates, while the phylogenetic trees obtained by combining datasets showed stable topologies. It is, therefore, more accurate to construct phylogenetic relationships for endophytic fungi based on multi-gene datasets. This study provides new insights into the distribution and characteristics of endophytic fungi in tobacco.

Fine needle aspiration of nodular cystic swelling showing a rare melanized fungus Medicopsis romeroi: A case report.

A young woman on long term corticosteroid therapy presented with a discrete hyperpigmented nodular cystic swelling on her upper thigh to the buttock region. There was history of discharging sinuses however at presentation the lesion was painless nondischarging. The lesion was subjected to fine-needle aspiration. The cytological smears on routine and special stains showed branching, septate fungal hyphae present extracellularly and intracellularly. The background showed dense mixed inflammation and granulomatous inflammation. The aspirated material was further subjected to culture. A nonsporulating melanized fungus was obtained and a culture isolate was sent for molecular characterization. Medicopsis romeroi, a rare melanized fungus belonging to the order Pleosporales was isolated on Internal transcribed spacer sequencing.

A new mixed inhibitor of adenosine deaminase produced by endophytic Cochliobolus sp. from medicinal plant seeds.

Medicinal plants have been studied for potential endophytic interactions and numerous studies have provided evidence that seeds harbor diverse microbial communities, not only on their surfaces but also within the embryo. Adenosine deaminase (ADA) is known as a potential therapeutic target for the treatment of lymphoproliferative disorders and cancer. Therefore, in this study, 20 types of medicinal plant seeds were used to screen endophytic fungi with tissue homogenate and streak. In addition, 128 morphologically distinct endophyte strains were isolated and their ADA inhibitory activity determined by a spectrophotometric assay. The strain with the highest inhibitory activity was identified as Cochliobolus sp. Seven compounds were isolated from the strain using a chromatography method. Compound 3 showed the highest ADA inhibitory activity and was identified as 5-hydroxy-2-hydroxymethyl-4H-pyran-4-one, based on the results of 1H and 13C NMR spectroscopy. The results of molecular docking suggested that compound 3 binds to the active site and the nonspecific binding site of the ADA. Furthermore, we found that compound 3 is a mixed ADA inhibitor. These results indicate that endophytic strains are a promising source of ADA inhibitors and that compound 3 may be a superior source for use in the preparation of biologically active ADA inhibitor compounds used to treat cancer.

Probe-Based Multiplex Real-Time PCR as a Diagnostic Tool to Distinguish Distinct Fungal Symbionts Associated With Euwallacea kuroshio and Euwallacea whitfordiodendrus in California.

California has been invaded by two distinct Euwallacea spp. that vector unique plant pathogenic symbiotic fungi on multiple hosts and cause Fusarium dieback. The objective of this study was to develop multiplex real-time quantitative PCR assays using hydrolysis probes targeting the ß-tubulin gene to detect, distinguish, and quantify fungi associated with the polyphagous shot hole borer (PSHB; Euwallacea whitfordiodendrus, Fusarium euwallaceae, Graphium euwallaceae, and Paracremonium pembeum) as well as the Kuroshio shot hole borer (KSHB; Euwallacea kuroshio, Fusarium kuroshium, and Graphium kuroshium) from various sample types. Absolute quantification reaction efficiencies ranged from 88.2 to 104.3%, with a coefficient of determination >0.992 and a limit of detection of 100 copies µl-1 for all targets across both assays. Qualitative detection using the real-time assays on artificially inoculated avocado shoot extracts showed more sensitivity compared with conventional fungal isolation from wood. All symbiotic fungi, except P. pembeum, from PSHB and KSHB female heads were detectable and quantified. Field samples from symptomatic Platanus racemosa, Populus spp., and Salix spp. across 17 of 26 city parks were positively identified as PSHB and KSHB through detection of their symbiotic fungi, and both were found occurring together on five trees from three different park locations. The molecular assays presented here can be utilized to accurately identify fungi associated with these invasive pests in California.

New species of Septoria associated with leaf spot diseases in Iran.

Species of Septoria are commonly associated with leaf spot diseases of a broad range of plant hosts worldwide. During our investigation of fungi associated with leaf spot diseases in northern and northwestern Iran, several Septoria isolates were recovered from symptomatic leaves on different herbaceous and woody plants in the Asteraceae, Betulaceae, and Salicaceae families. These isolates were studied by applying a polyphasic approach including morphological and cultural data and a multigene phylogeny using a combined data set of partial sequences of the 28S nuc rRNA gene (large subunit [28S]), internal transcribed spacer regions and intervening 5.8S nuc rRNA gene (ITS) of the nuc rDNA operon, actin (actA), translation elongation factor 1-α (tef1), calmodulin (cmdA), ß-tubulin (tub2), and DNA-directed RNA polymerase II second largest subunit (rpb2). Four novel species are proposed, namely, Septoria eclipticola on Eclipta prostrata, Septoria firouraghina on Cirsium arvense, Septoria guilanensis on Populus deltoides, and Septoria taleshana on Alnus subcordata. All species are illustrated, and their morphology and phylogenetic relationships with other Septoria species are discussed.

Subcutaneous Fungal Infection Caused by a Non-sporulating Strain of Corynespora cassiicola Successfully Treated with Terbinafine.

Corynespora cassiicola is a common plant pathogen, but C. cassiicola infection in human hosts is extremely rare. In this report, we present an 84-year-old male with long-term use of inhaled corticosteroids who developed a subcutaneous infection caused by C. cassiicola. The organism was isolated from both wound culture and biopsy specimen from the skin lesion. However, no microscopic diagnostic characters could be obtained because the isolates failed to sporulate on different culture media. Molecular diagnosis by amplification and sequencing of the internal transcribed spacer regions of ribosomal DNA was performed, and the sequences of the isolates were identical to those of C. cassiicola. The patient was treated successfully with oral terbinafine therapy for 12 weeks. In this report, we also review the epidemiology, clinical and therapeutic facets of cutaneous C. cassiicola infection.

Multilocus phylogeny of Acrospermaceae: New epibiotic species and placement of Gonatophragmium, Pseudovirgaria, and Phaeodactylium anamorphs.

Acrospermum is a poorly known genus of epibiotic and saprophytic species with a subcosmopolitan distribution. Here, we investigate the intriguing relationship between Acrospermum and its host plants in the fern family Polypodiaceae, where it occurs upon approximately 45 neotropical species. We conducted phylogenetic analyses using an eight-marker comprehensive ascomycete data set comprising 719 species representing all major lineages along with 23 new Acrospermum specimens sampled from ferns. We ask whether fern-dwelling Acrospermum are monophyletic, whether epibiotic Acrospermum have evolved independently from saprophytic ancestors, and identify anamorphic phases by incorporating sequences for all suspected taxa. Our results corroborate the placement of Acrospermales within the Dothideomycetes with strong support. However, the order remains incertae sedis due to weak support along the branches subtending the clade that includes the Acrospermales plus Dyfrolomycetales. Our results show a strong phylogenetic pattern in lifestyles but do not clearly identify an ancestral life history state. The first divergence in Acrospermaceae splits fungicolous taxa from taxa that inhabit plants; saprophytes and anamorphic phases found on angiosperms occur in both clades. Fungicolous species are monophyletic, whereas species with an epibiotic or necrotic life history upon plants are nonmonophyletic due to the position of the saprophyte A. longisporium. Previously, all Acrospermum collected from ferns were identified as A. maxonii. Our results indicate that this is not monophyletic due to the inclusion of Gonatophragmium triuniae. Two species are described herein as A. gorditum, sp. nov., and A. leucocephalum, sp. nov. We find no instances of co-cladogenesis; however, our ability to detect this is limited by the lack of resolution in the A. maxonii clade. Rather, we see that that the distribution of epibiotic Acrospermum is explained by the overlap between the ecological niche of the Acrospermum species and its host.

Long-Term Carriage of Medicopsis romeroi, an Agent of Black-Grain Mycetoma, Presenting as Phaeohyphomycosis in a Renal Transplant Patient.

Medicopsis species are rare fungal pathogens that frequently resist common antifungal therapies and are difficult to identify morphologically as conidia are produced in pycnidia, a key feature of coelomycetes. Immunocompromised patients are at risk of these infections, even after remote exposure, and typically present with phaeohyphomycoses without dissemination. We present the case of a renal transplant recipient 6.5 years post-transplant who developed a slowly progressive soft tissue infection mimicking a synovial cyst. A cultured isolate was identified as Medicopsis romeroi by sequencing of multiple ribosomal loci. The patient responded well to debridement and posaconazole therapy. Solid-organ transplant patients are at risk of opportunistic fungal infection long after transplant, and molecular methods are often required for definitive identification.

Vochysiamides A and B: Two new bioactive carboxamides produced by the new species Diaporthe vochysiae.

Endophytic fungi have been considered a rich source for bioactive secondary metabolites with novel chemical structures. A high diverse group of endophytes, isolated from different medicinal plants, belongs to the genus Diaporthe. In a previously study performed by our group the crude extract of strain LGMF1583 showed considerable antibacterial activity mainly against Gram-negative bacteria. Based on ITS phylogeny analysis, strain LGMF1583 was identified as belonging to Diaporthe genus and may represent a new species. In the present study, we described the new species Diporthe vochysiae based on multilocus phylogeny analysis and morphological characteristics. The species name refers to the host, from which strain LGMF1583 was isolated, the medicinal plant Vochysia divergens. In view of the biotechnological potential of strain LGMF1583, we have also characterized the secondary metabolites produced by D. vochysiae. Chemical assessment of the D. vochysiae LGMF1583 revealed two new carboxamides, vochysiamides A (1) and B (2), in addition to the known metabolite, 2,5-dihydroxybenzyl alcohol (3). In the biological activity analysis, vochysiamide B (2) displayed considerable antibacterial activity against the Gram-negative bacterium Klebsiella pneumoniae (KPC), a producer of carbapenemases, MIC of 80 µg/mL. Carbapenemases are considered a major antimicrobial resistance threat, and infections caused by KPC have been considered a public health problem worldwide, and new compounds with activity against this bacterium are nowadays even more required.

A new genus, Zhurbenkoa, and a novel nutritional mode revealed in the family Malmideaceae (Lecanoromycetes, Ascomycota).

Lichen-inhabiting fungi are highly specialized mycoparasites, commensals or rarely saprotrophs, that are common components of almost every ecosystem, where they develop obligate associations with lichens. Their relevance, however, contrasts with the relatively small number of these fungi described so far. Recent estimates and ongoing studies indicate that a significant fraction of their diversity remains undiscovered and may be expected in tropical regions, in particular in hyperdiverse fog-exposed montane forests. Here, we introduce the new genus Zhurbenkoa, from South America and Europe, for three lichenicolous fungi growing on thalli of the widespread lichen genus Cladonia (Lecanorales). Phylogenetic analyses based on combined sequence data of mt and nuc rDNA obtained from Andean populations (Bolivia) placed Zhurbenkoa as a member of Malmideaceae, a recently introduced family of lichen-forming fungi in the class Lecanoromycetes. Zhurbenkoa is closely related to the genera Savoronala and Sprucidea. The new genus is characterized by the development of grayish brown to almost black apothecia lacking an evident margin, an epihymenium interspersed with crystals (often seen as pruina), a strongly conglutinated hymenium made of noncapitate and sparsely branched paraphyses, a colorless exciple composed of radially arranged hyphae, a Lecanora/Micarea-like ascus type, and aseptate or 1-septate ellipsoidal colorless ascospores. Zhurbenkoa includes two Neotropical (Z. cladoniarum, Z. latispora) and one widespread (Z. epicladonia) species. The lichenicolous trophic mode is documented for the first time in the Malmideaceae, which until now included only lichen-forming associations between fungi and green algae.

Bioactive exopolysaccharide from Neopestalotiopsis sp. strain SKE15: Production, characterization and optimization.

Fungal exopolysaccharides are powerful resources of medicinal applications. Neopestalotiopsis sp. SKE15 was isolated and identified according to phenotypical and genotypical analyses (GenBank Accession No. MG649986). The exopolysaccharide (EPS) was produced by cultivation of mycelia in broth culture and extracted. The production was optimized to 2.02 g/l after selection of agitation, temperature, FeSO4 and K2HPO4 concentrations as the most influencing factors using Placket-Burman design and then by applying response surface methodology. Analytical Tools showed that the EPS is composed of a polysaccharide (1.5-2.1 × 106 Da) and its probable low molecular weight derivatives, in a wide range of chain lengths, among them an oligosaccharide of about 1970 Da was dominant. GC-MS (Gas chromatography-mass spectrometry) analysis revealed the EPS was mainly constructed from d-glucose, sorbitol and D-galactose. The EPS showed antibacterial activity against Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 6633 and Pseudomonas aeruginosa ATCC 27853. DPPH (1,1-diphenyl-2-picryl-hydrazyl) and hydroxyl radical scavenging activity assays showed strong antioxidant activity of the EPS. A challenge with three different cancerous cell lines showed cytotoxic activity of the EPS at final concentration of 100 and 200 µg/ml. Further investigation on medicinal applications of the biopolymer is promising.