Antiproliferative and Antibacterial Activity of Polyporoid Fungi from Veracruz, Mexico.

Polyporoid fungi represent a vast source of bioactive compounds with potential pharmacological applications. The importance of polyporoid fungi in traditional Chinese medicine has led to an extensive use of some species of Ganoderma for promoting health and longevity because their consumption is associated with several bioactivities. Nevertheless, bioactivity of some other members of the Polyporaceae family has also been reported. This work reports the antiproliferative and antibacterial activity of crude extracts obtained from fruiting bodies of polypore fungi collected from the central region of Veracruz, Mexico, aimed at understanding the diversity of polypore species with potential pharmacological applications. 29 collections were identified macro- and microscopically in 19 species of polyporoid fungi, belonging to 13 genera. The antiproliferative activity screening of extracts against solid tumor cell lines (A549, SW1573, HeLa, HBL-100, T-47D, WiDr) allow us to identify four extracts with strong bioactivity [half-maximal growth inhibition (GI50) ≤ 50 µg/mL]. After this, a phylogenetic analysis of DNA sequences from the ITS region obtained from bioactive specimens allowed us to identify three extracts as Pycnoporus sanguineus (GI50 = ≤ 10 µg/mL) and the fourth bioactive extract as Ganoderma oerstedii (GI50 = < 50 µg/mL. Likewise, extracts from P. sanguineus showed mild or moderate antibacterial activity against Escherichia coli, Staphylococcus aureus and Xanthomonas albilineas. Bioprospecting studies of polyporoid fungi add to the knowledge of the diversity of macrofungi in Mexico and allow us to select one of the bioactive P. sanguineus to continue the pursuit of bioactive compounds through mycochemical studies.

Taxonomy of the major rhizomorphic species of the "Melanopus group" within Polyporaceae in Yasuní National Park, Ecuador.

Yasuní National Park in Ecuador is one of the most biodiverse places on earth. The fungi in this tropical rainforest are also diverse but have received little research attention. This research paper focuses on an important group of fungi in the family Polyporaceae and examines the genera Polyporus, Atroporus, and Neodictyopus that form aerial melanized cord-like structures called rhizomorphs. Phylogenetic analyses, macro and micromorphological descriptions of basidiomata and rhizomorphs, as well as cultural characterization were completed to better understand these ecologically important fungi. Here we describe four new species: Atroporus yasuniensis, Atroporus tagaeri, Neodictyopus sylvaticus, and Polyporus taromenane, and a new variety Polyporus leprieurii var. yasuniensis. The information presented in this study adds important new knowledge about the unusual rhizomorph producing fungi found in Yasuní National Park, Ecuador and other tropical rainforests.

Genome sequencing and annotation and phylogenomic analysis of the medicinal mushroom Amauroderma rugosum, a traditional medicinal species in the family Ganodermataceae.

Amauroderma rugosum is one of the traditional Chinese medicinal mushrooms and is used to reduce inflammation, treat diuretic and upset stomach, and prevent cancer. Here, we present a genomic resource of Amauroderma rugosum (ACCC 51706) for further understanding its biology and exploration of the synthesis pathway of bioactive compounds. Genomic DNA was extracted and then subjected to Illumina HiSeq X Ten and PacBio Sequel I sequencing. The final genome is 40.66 Mb in size, with an N50 scaffold size of 36.6 Mb, and encodes 10 181 putative predicted genes. Among them, 6931 genes were functionally annotated. Phylogenomic analysis suggested that A. rugosum and Ganoderma sinense were not clustered together into a group and the latter was grouped with the Polyporaceae. Further, we also identified 377 carbohydrate-active enzymes (CAZymes) and 15 secondary metabolite biosynthetic gene clusters. This is the first genome-scale assembly and annotation for an Amauroderma species. The identification of novel secondary metabolite biosynthetic gene clusters would promote pharmacological research and development of novel bioactive compounds in the future.

Species Prioritization Based on Spectral Dissimilarity: A Case Study of Polyporoid Fungal Species.

Biological species collections are critical for natural product drug discovery programs. However, prioritization of target species in massive collections remains difficult. Here, we introduce an untargeted metabolomics-based prioritization workflow that uses MS/MS molecular networking to estimate scaffold-level distribution. As a demonstration, we applied the workflow to 40 polyporoid fungal species. Nine species were prioritized as candidates based on the chemical structural and compositional similarity (CSCS) metric. Most of the selected species showed relatively higher richness and uniqueness of metabolites than those of the others. Cryptoporus volvatus, one of the prioritized species, was investigated further. The chemical profiles of the extracts of C. volvatus culture and fruiting bodies were compared, and it was shown that derivative-level diversity was higher in the fruiting bodies; meanwhile, scaffold-level diversity was similar. This showed that the compounds found from a cultured fungus can also be isolated in wild mushrooms. Targeted isolation of the fruiting body extract yielded three unknown (1-3) and six known (4-9) cryptoporic acid derivatives, which are drimane-type sesquiterpenes with isocitric acid moieties that have been reported in this species. Cryptoporic acid T (1) is a trimeric cryptoporic acid reported for the first time. Compounds 2 and 5 exhibited cytotoxicity against HCT-116 cell lines with IC50 values of 4.3 and 3.6 µM, respectively.

GC-MS Analysis and Cytotoxicity Detection of Volatile Oil from the Fragrant Bracket Mushroom, Trametes suaveolens (Agaricomycetes).

Trametes suaveolens is a medicinal mushroom known as Baizhi in traditional Chinese medicine. Our previous research has found that it has some pharmacological activity in vivo. The aim of the study was to investigate the chemical compounds and cytotoxic effects of volatile oil from T. suaveolens. In this study, internal transcribed spacer (ITS) sequence analysis was used to determine wild T. suaveolens collected. To fully analyze the composition of volatile oil extracted from T. suaveolens, hydrodistillation (HD) and solid phase microextraction (SPME) were adopted simultaneously. In both cases, the analysis was carried out using gas chromatography-mass spectrometry (GC-MS) and the cytotoxic effects of T. suaveolens volatile oil on human NCI-H460 lung non-small cell carcinoma cells and MCF-7 breast adenocarcinoma cells were investigated. The results indicated that all these wild samples were identified as T. suaveolens. Thirty-one components in HD and 62 components in SPME were identified, respectively. Furthermore, the volatile compounds obtained from T. suaveolens by HD indicated that esters compounds were a major class (68.47%), followed by acids (25.06%), aldehydes (4.20%), and alcohols (1.48%). SPME found that the largest content were aldehydes (45.47%), followed by alcohols (31.42%), ketones (6.89%), and esters (6.72%). In the cytotoxic assays, the volatile oil was found to have toxic effect on NCI-H460 and MCF-7 tumor cells but not BEAS-2B and MCF-10A normal cells, and the IC50 values of NCI-H460 and MCF-7 tumor cells were 24.1 and 19.2 µg/ml, respectively. The present study shows that the composition of essential oil from T. suaveolens has potential value for the prevention and treatment of lung cancer.

Genome and secretome analysis of jute endophyte Grammothele lineata strain SDL-CO-2015-1: Insights into its lignocellulolytic structure and secondary metabolite profile.

Grammothele lineata strain SDL-CO-2015-1, jute (Corchorus olitorius) endophyte has been reported to produce anti-cancer drug paclitaxel in culture condition. Here we investigated the genome using different bioinformatic tools to find its association with the production of commercially important compounds including taxol. Carbohydrate-active enzymes, proteases, and secretory proteins were annotated revealing a complex endophytic relationship with its plant host. The presences of a diverse range of CAZymes including numerous lignocellulolytic enzymes support its potentiality in biomass degradation. Genome annotation led to the identification of 28 clusters for secondary metabolite biosynthesis. Several biosynthesis gene clusters were identified for terpene biosynthesis from antiSMASH analysis but none could be specifically pinned to taxol synthesis. This study will direct us to understand the genomic organization of endophytic basidiomycetes with a potential for producing numerous commercially important enzymes and secondary metabolites taking G. lineata as a model.

Notes on Medicinal Polypore Species from the Genus Daedaleopsis (Agaricomycetes), Distributed in the Asian Part of Russia.

We present a study on the medicinal value, taxonomy, and ecology of the polypore mushrooms Daedaleopsis confragosa and D. tricolor isolated from the Asian part of Russia (the Urals, Siberia, and the Far East). The phylogenetic analysis of recombinant DNA internal transcribed spacer sequences data has shown that D. confragosa and D. tricolor do not differ taxonomically and should be considered as one species. However, because D. confragosa and D. tricolor differ significantly in their ecological characteristics, they may be considered as two morpho-ecological varieties: D. confragosa var. confragosa and D. confragosa var. tricolor (both nomen provisiorum).

Profiles of Little-Known Medicinal Polypores: Haploporus odorus (Agaricomycetes).

The purpose of this study was to compile a comprehensive characterization of little-known polypores, which have recently been found to possess anticancer activity and thus can also be used in cancer target therapy. Haploporus odorus is a polypore of Holarctic distribution and has been found by harvesters working in taiga floodlands and broadleaf forests of the Northern Hemisphere. A substance known as haploporic acid A was determined in methylene chloride extract from the dried basidiomata of H. odorus. This substance can be used in cancer therapy; more details of its health benefits could be used in mycotherapy.

Purification and characterization of a novel extracellular neutral metalloprotease from Cerrena albocinnamomea.

We selected a fungus secreting a neutral protease from soil and identified it as the basidiomycete fungus Cerrena albocinnamomea according to its ITS-5.8S rDNA and 28S rDNA-D1/D2 sequences. A major extracellular protease isolated from C. albocinnamomea was purified approximately 44-fold through two purification steps. SDS-PAGE analyses of the purified protease revealed a single band, and its molecular mass of 39,756 Da was determined using MALDI-TOF-MS. The enzyme was optimally active at approximately pH 7.0 and 45°C. The Km and Vmax values for the hydrolysis of azocasein were 2.46 mg/mL and 989 units/min/mg protein, respectively. The enzyme was stable at pH 3.6-8.6 for 16 h and at temperatures ≤35°C for 1 h. Enzymatic activity was completely inhibited by Cu2+ and Zn2+ and markedly by EDTA and phosphoramidon. The N-terminal amino acid sequence ASYRVLPIT is highly similar to those of the members of the metalloprotease family M36, such as keratinase and elastinase. However, the protease did not detectably hydrolyze keratin or elastin. In contrast, the protease hydrolyzed fibrinogen, although there were no significant sequence similarities to the N-terminal amino acid sequences of other fibrinolytic enzymes. These results suggest that the purified protease represents a new neutral metalloprotease with fibrinogenolytic activity.

Phylogeny and taxonomy of Favolus (Basidiomycota).

Favolus is a natural genus that is closely related to Polyporus. A taxonomic study of Favolus was carried out based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2 = ITS) and 18S and 28S genes of the nuc rDNA, translation elongation factor 1-α gene (TEF1-α), the mt 16S rDNA gene (mt rrnS), ß-tubulin gene (TBB1), and the RNA polymerase II largest subunit (RPB1) and second largest subunit (RPB2) genes. Fourteen specimens collected from subtropical to tropical regions in China were shown to be different from existing Favolus species. They are described and illustrated here as four new species, namely, Favolus niveus, F. septatus, F. pseudoemerici, and F. subtropicus.

Lentinoid and Polyporoid Fungi, Two Generic Conglomerates Containing Important Medicinal Mushrooms in Molecular Perspective.

Polyporoid and lentinoid fungi contain the important producers of substances having immunomodulatory, antitumoral, antiviral, and antihyperlipidemic effects. The discovery of several phylogenetic lines within the lentinoid-polyporoid continuum will help with target metabolomic analysis of species still not studied in pharmacological respects. The purpose of the present work was to increase a resolution in the lentinoid-polyporoid phylogenetic zone by means of selection of both the main representatives of Lentinus-related genera and poorly known/intermediate taxa such as Lentinus suavissimus, Neofavolus spp., and the resupinate part of Polyporus (genera Perenniporia and Pachykytospora) in the context of the basic structure of the Polyporales tree. The molecular phylogeny of highlighting all the polyporoid and lentinoid nodes was reconstructed using nLSU ITS rDNA and TEF datasets. The data obtained from ITS, TEF, and LSU coincide in support of core Polyporaceae of 10 clades corresponded to the generic level and 7 of these (Cerioporus, Cladomeris, Favolus, Lentinus, Neofavolus, Picipes, and Polyporus s.str.) contain generic units characterized by polyporoid or lentinoid morphotypes. The other 2 clades containing lentinoid taxa are outside the core Polyporaceae, namely Panus (Meruliaceae, Polyporales) and Neolentinus (Gloeophyllaceae, Gloeophyllales). A new genus, Picipes, is described and 25 new combinations are proposed.

Ethnomedicinal uses, pharmacological activities, and cultivation of Lignosus spp. (tiger׳s milk mushrooms) in Malaysia - A review.

ETHNOPHARMACOLOGICAL RELEVANCE: Several members of the genus Lignosus, which are collectively known as cendawan susu rimau (in Malay) or tiger׳s milk mushrooms (TMM), are regarded as important local medicine particularly by the indigenous communities in Malaysia. The mushroom sclerotia are purportedly effective in treating cancer, coughs, asthma, fever, and other ailments. The most commonly encountered Lignosus spp. in Malaysia was authenticated as Lignosus rhinocerotis (Cooke) Ryvarden (synonym: Polyporus rhinocerus), which is also known as hurulingzhi in China and has been used by Chinese physicians to treat liver cancer, gastric ulcers, and chronic hepatitis. In spite of growing interest in the therapeutic potential of TMM, there is no compilation of scientific evidence that supports the ethnomedicinal uses of these mushrooms. Therefore, the present review is intended (i) to provide a comprehensive, up-to-date overview of the ethnomedicinal uses, pharmacological activities, and cultivation of TMM in general and L. rhinocerotis in particular, (ii) to demonstrate how recent scientific findings have validated some of their traditional uses, and (iii) to identify opportunities for future research and areas to prioritize for TMM bioprospecting. MATERIALS AND METHODS: A detailed literature search was conducted via library search (books, theses, reports, newspapers, magazines, and conference proceedings) and electronic search (Web of Science, PubMed, and Google Scholar) for articles published in peer-reviewed journals. These sources were scrutinized for information on TMM and specifically for L. rhinocerotis. RESULTS: Ethnomycological knowledge about TMM, with an emphasis on cultural associations and use as local medicine, has been comprehensively and systematically compiled for the first time. Some of the reported medicinal properties of TMM have been validated by scientific studies. The anti-tumor, immuno-modulatory, anti-inflammatory, anti-oxidative, anti-microbial, neurite outgrowth stimulation, and other pharmacological activities of L. rhinocerotis sclerotial extracts have been explored. The nature of sclerotial bioactive components, such as proteins, polysaccharides, and/or polysaccharide-protein complexes, has been identified, whereas the low-molecular-weight constituents remain poorly studied. The artificial cultivation of L. rhinocerotis via solid substrate and liquid fermentations successfully yielded fruiting bodies, sclerotium, mycelium, and culture broth that could be exploited as substitutes for the wild resources. The cultivated sclerotium and mycelium were shown to be safe from a toxicological point of view. Other research areas, e.g., chemical studies, genomics, and proteomics, have been employed to gain insights into the medicinal properties of TMM. CONCLUSIONS: This review clarified the medicinal properties of TMM as recorded in various ethnomycological records, and it simultaneously highlighted the current efforts to provide scientific evidence by using various in vitro and in vivo models. Thus far, only the anti-tumor and immuno-modulatory effects of L. rhinocerotis sclerotial aqueous extracts have been extensively investigated, and other medicinal properties relevant to their traditional uses, e.g., anti-tussive and anti-pyretic properties, have yet to be validated. Further studies focusing on (i) the isolation and characterization of active components, (ii) the elucidation of their modes of action, and (iii) an evaluation of their safety and efficacy, when compared with the crude aqueous preparations, are warranted to accelerate potential drug discovery from TMM.

Phylogenetic relationships and morphological evolution in Lentinus, Polyporellus and Neofavolus, emphasizing southeastern Asian taxa.

The genus Lentinus (Polyporaceae, Basidiomycota) is widely documented from tropical and temperate forests and is taxonomically controversial. Here we studied the relationships between Lentinus subg. Lentinus sensu Pegler (i.e. sections Lentinus, Tigrini, Dicholamellatae, Rigidi, Lentodiellum and Pleuroti and polypores that share similar morphological characters). We generated sequences of internal transcribed spacers (ITS) and partial 28S regions of nuc rDNA and genes encoding the largest subunit of RNA polymerase II (RPB1), focusing on Lentinus subg. Lentinus sensu Pegler and the Neofavolus group, combined these data with sequences from GenBank (including RPB2 gene sequences) and performed phylogenetic analyses with maximum likelihood and Bayesian methods. We also evaluated the transition in hymenophore morphology between Lentinus, Neofavolus and related polypores with ancestral state reconstruction. Single-gene phylogenies and phylogenies combining ITS and 28S with RPB1 and RPB2 genes all support existence of a Lentinus/Polyporellus clade and a separate Neofavolus clade. Polyporellus (represented by P. arcularius, P. ciliatus, P. brumalis) forms a clade with species representing Lentinus subg. Lentinus sensu Pegler (1983), excluding L. suavissimus. Lentinus tigrinus appears as the sister group of Polyporellus in the four-gene phylogeny, but this placement was weakly supported. All three multigene analyses and the single-gene analysis using ITS strongly supported Polyporus tricholoma as the sister group of the Lentinus/Polyporellus clade; only the 28S rRNA phylogeny failed to support this placement. Under parsimony the ancestral hymenophoral configuration for the Lentinus/Polyporellus clade is estimated to be circular pores, with independent transitions to angular pores and lamellae. The ancestral state for the Neofavolus clade is estimated to be angular pores, with a single transition to lamellae in L. suavissimus. We propose that Lentinus suavissimus (section Pleuroti) should be reclassified as Neofavolus suavissimus comb. nov.

Molecular phylogeny and morphology reveal a new species of Amyloporia (Basidiomycota) from China.

Amyloporia pinea sp. nov. is described and illustrated on the basis of collections from southern China. Morphology and phylogenetic analysis of rDNA ITS sequences support this new species. Morphologically, it is characterized by resupinate, annual basidiocarps, cream to yellowish buff pore surface when fresh, which becomes yellowish brown to clay-buff upon drying, a dimitic hyphal system with clamped generative hyphae and inamyloid skeletal hyphae, fusoid cystidioles, and cylindrical basidiospores; moreover, it causes a brown rot. Molecular phylogeny inferred from ITS sequence data suggested a close relationship between A. pinea and Amyloporia crassa sensu lato. Antrodia subxantha has amyloid skeletal hyphae, and grouped within the Amyloporia clade, hence, it is transferred to Amyloporia, and a new combination Amyloporia subxantha is proposed.

Cytotoxic metabolites from Perenniporia tephropora, an endophytic fungus from Taxus chinensis var. mairei.

Based on bioactivity-oriented isolation, the EtOAc extract of a culture broth of the endophytic fungus Perenniporia tephropora Z41 from Taxus chinensis var. mairei, with strong anti-Pyricularia oryzae activity, afforded a new sesquiterpenoid, perenniporin A (1), together with three known compounds, ergosterol (2), rel-(+)-(2aR,5R,5aR,8S,8aS,8bR)-decahydro-2,2,5,8-tetramethyl-2H-naphtho[1,8-bc]genfuran-5-ol (3), and albicanol (4). Their structures were elucidated by means of spectroscopic methods. All the isolated compounds and the EtOAc extract of P. tephropora Z41 (EPT) were evaluated for their cytotoxic activity against three human cancer cell lines (HeLa, SMMC-7721, and PANC-1). EPT demonstrated significant cytotoxicity with IC(50) values ranging from 2 to 15 µg/mL. Compound 2 was the most cytotoxic constituent against the tested cell lines with IC(50) values of 1.16, 11.63, and 11.80 µg/mL, respectively, while compounds 1, 3, and 4 exhibited moderate cytotoxicity with IC(50) values ranging from 6 to 58 µg/mL. We conclude that the endophytic fungus P. tephropora is a promising source of novel and cytotoxic metabolites.

First human case of pulmonary fungal ball due to a Perenniporia species (a basidiomycete).

Perenniporia species are basidiomycetes, resupinate shelf fungi responsible for white rot decay of wood. Here, we report for the first time an intracavitary pulmonary fungal ball due to a species of Perenniporia that has not been recognized so far as a human pathogen. The fungus was identified by sequencing of the partial ribosomal operon of a culture from a clinical specimen.

[Polypore species diversity, floral composition, and distribution characteristics in Changbai Mountains, Northeast China].

Polypore is an important group of wood-rotting fungi, which can decompose wood cellulose, semi-cellulose, and lignin into nutrients to be utilized by itself and other organisms, and accordingly, can promote the material cycling in forest ecosystem. Based on more than 10 years investigation and collection, it was found that the polypore in Changbai Mountains was rich in species diversity, with the Shannon diversity index being 5.06. In the Mountains, a total of 246 polypore species were recorded, occupying 40.7% of the total in China. The polypore species recorded belonged to 80 genera, 11 families, and 6 orders, among which, Polyporaceae was the dominant family. The most important bio-geographical elements of the polypore in Changbai Mountains were north temperate element and cosmopolitan element, other elements also included, showing a distinct north temperate character in floral composition. Pinus was the main host of polypore, supporting 41.5% of the polypore in Changbai Mountains. Most of the polypore was saprophytic, and its quantity had a significant correlation with the decaying degree of substrates, implying its important role in the material cycling in forest ecosystem in Changbai Mountains. There were 18 threatened polypore species in the Mountains, accounting for 37.5% of total in China. Besides their important ecological functions, many of the polypore also had higher economic value. On the premise of complete protection on them, these fungal resources should be exploited reasonably.

Importance of natural disturbance for recovery of the rare polypore Antrodiella citrinella Niemelä & Ryvarden.

Most strictly protected areas world-wide have been established in former commercial forests, but there is little evidence that this strongly advocated tool of conservation has had positive effects on endangered species in the short run of decades. One of the major reasons for the failure of habitat improvement in most of these reserves is a lack of natural disturbance. Consequently, only a few studies of species recovery in protected areas after natural disturbance are available. We demonstrate how natural dynamics in the oldest national park in Germany allows the area-wide recovery of the rare wood-inhabiting fungus Antrodiella citrinella. Our analyses showed that an increase of dead wood at >134-224 m(3)ha(-1), amounts known from primeval forests, allowed the fungus, which has barely survived as 'living dead' in two remnants of pristine forests, to spread from this small relict populations to the whole national park, within a radius of more than 30 km. Our result demonstrate that natural disturbance, even in former commercial forests, allowed the recovery of this rare species and fully supports the use of passive management in protected areas, even if natural dynamics often evokes debates on salvage logging.

Relationships among North American and Japanese Laetiporus isolates inferred from molecular phylogenetics and single-spore incompatibility reactions.

Relationships were investigated among North American and Japanese isolates of Laetiporus using phylogenetic analysis of ITS sequences and single-spore isolate incompatibility. Single-spore isolate pairings revealed no significant compatibility between North American and Japanese isolates. ITS analysis revealed 12 clades within the core Laetiporus clade, seven of which are known to occur in North America (including Hawaii and the Caribbean), three in Japan, two in South America, three in Europe and one in South Africa. The identity of L. sulphureus s.s. has yet to be determined and could be either L. "sulphureus" (clade C), which appears to be restricted to Europe and occurs on angiosperms and gymnospersm, or L. "sulphureus" (clade E), which is found in Europe, North America and South America exclusively on angiosperms. Three clades, one from the Caribbean, one from Hawaii and one from South Africa, have yet to be named formally. Of the three Laetiporus species found in Japan two have been named recently (L. cremeiporus and L. montanus) and one has been epitypified (L. versisporus). The single-spore incompatibility and ITS data support recognition of the three Japanese taxa as distinct biological and evolutionary species.

Cerrena aurantiopora sp. nov. (Polyporaceae) from eastern Asia.

A new species of polypore in genus Cerrena was discovered in Kangwon Province, Korea. The resupinate basidiocarp and light orange, poroid hymenophore were sufficiently different to be distinguished from previously recorded species of Cerrena, C. consors, C. cystidiata, C. sclerodepsis and C. unicolor. Based on the results of morphological and phylogenetic analyses, we propose this new polypore as Cerrena aurantiopora sp. nov.