Screening of White-Rot Fungi Isolates for Decolorization of Pulp and Paper Mill Effluent and Assessment of Biodegradation and Biosorption Processes.

Ten white-rot fungal isolates were evaluated for the decolorization potential of pulp and paper mill effluent. Trametes elegans PP17-06, Pseudolagarobasidium sp. PP17-33, and Microporus sp.2 PP17-20 showed the highest decolorization efficiencies between 42 and 54% in 5 d. To reveal the mechanisms involved in decolorization and assess the long-term performance, PP17-06, which showed the highest decolorization efficiency, was further investigated. It could reduce the ADMI color scale by 63.6% in 10 d. However, extending the treatment period for more than 10 d did not significantly enhance the decolorization efficiencies. The maximum MnP activity of 3.27 U L-1 was observed on the 6 d during the biodegradation. In comparison, laccase activities were low with the maximum activity of 0.38 U L-1 (24 d). No significant LiP activities were monitored during the experiment. Dead fungal biomass showed an optimum decolorization efficiency of 44.18% in 8 d employing the biosorption mechanism. No significant changes in the decolorization efficiency were observed after that, suggesting the equilibrium status was reached. These results revealed that PP17-06 has the potential to decolorize pulp and paper mill effluent by employing both biodegradation and biosorption processes.

Nutritional and Biochemical Characterization of Panus lecomtei Mushroom (Agaricomycetes) from India and Its Cultivation.

Panus lecomtei is emerging as an edible mushroom found worldwide and particularly in the Northern Hemisphere. The mushroom contains a substantial amount of useful nutritional and medicinal compounds. In the present study, we have examined a specimen of P. lecomtei submitted to the ICAR-Directorate of Mushroom Research gene bank. The specimen was examined for taxonomical characters using classical and molecular tools. Attempts were made for cultivation of this mushroom under controlled conditions using sawdust-based substrate. The specimen was characterized by its purplish fruiting body having coarse, rigid, dense hairs on the cap, pubescent stipe, and abundant metuloids. Molecular identification through conserved ITS region was done and the sequence was deposited in NCBI GenBank under accession number MN332200. Nutritional profiling and biochemical analysis showed that the mushroom contained high carbohydrate but low fat contents. The mushroom showed the presence of phenolics, ß-carotene, and lycopene. The analysis also showed substantial antioxidant properties in the mushroom. The findings presented herein point out that P. lecomtei can be used as a potential edible mushroom for diversification of mushroom production in India.

An overview of Antrodiella and related genera of Polyporales from the Neotropics.

The genus Antrodiella includes resupinate and pileate species of polypores with a dimitic hyphal system, small, globose to cylindrical basidiospores, absence of cystidia, tetrapolar mating system, and haplo-dikaryotic nuclear behavior. Recent studies, however, indicate that Antrodiella is highly polyphyletic, so many of its species have been transferred to other genera. This study reviews the systematic status and diversity of Antrodiella from the Neotropics based, in part, on studies of type specimens. Collections from Brazil were used for molecular analysis of nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS), nuc 28S rDNA (28S), and portions of genes encoding translation elongation factor 1-α (tef1) and the second largest subunit of RNA polymerase II (rpb2). Eight genera are confirmed to include Neotropical species treated as Antrodiella in a broad sense: Aegis, Antrodiella s. str., Flaviporus, Metuloidea, Mycorrhaphium, Rickiopora, Trametopsis, and Trullella. Molecular data reveal the occurrence of two new species, described as Antrodiella trivialis, the only Neotropical species of Antrodiella s. str. known so far, and Mycorrhaphium hispidum. In addition, Antrodiella luteocontexta was found to nest in the genus Aegis, close to the Grifolaceae and Polyporaceae; therefore, the new combination Aegis luteocontexta is proposed. Comments on the eight Antrodiella-related genera as well as species with uncertain taxonomic position are provided, together with a key to their identification.

Skeletocutins A-L: Antibacterial Agents from the Kenyan Wood-Inhabiting Basidiomycete, Skeletocutis sp.

Fermentation of the fungal strain Skeletocutis sp. originating from Mount Elgon Natural Reserve in Kenya, followed by bioassay guided fractionation led to the isolation of 12 previously undescribed metabolites named skeletocutins A-L (1-5 and 7-13) together with the known tyromycin A (6). Their structures were assigned by NMR spectroscopy complemented by HR-ESIMS. Compounds 1-6 and 11-13 exhibited selective activities against Gram-positive bacteria, while compound 10 weakly inhibited the formation of biofilm of Staphylococcus aureus. The isolated metabolites were also evaluated for inhibition of L-leucine aminopeptidase, since tyromycin A had previously been reported to possess such activities but only showed weak effects. Furthermore, all compounds were tested for antiviral activity against Hepatitis C virus (HCV), and compound 6 moderately inhibited HCV infectivity with an IC50 of 6.6 µM.

Trunk surface agarwood-inducing technique with Rigidoporus vinctus: An efficient novel method for agarwood production.

Only when Aquilaria spp. or Gyrinops spp. trees are wounded, due to insect attack, or microbial invasion, agarwood can be successfully induced. In the present study, a fungus which can induce agarwood formation efficiently was isolated and a suitable method for its application to induce agarwood formation was developed. Rigidoporus vinctus was isolated from the inner layers from infectious A. sinensis trees. When the fermentation liquid of fungi inoculated back to A. sinensis tree, agarwood was found to be induced. In addition, a novel method called trunk surface agarwood-inducing technique (Agar-Sit) was developed to produce agarwood with R. vinctus. The alcohol soluble extract content of the agarwood, up to 38.9%, far higher than the requirement (10%) in Chinese Pharmacopoeia and the six characteristic compounds of agarwood used as Chinese Medicinal Materials were all detected. Their relative percentages of the sesquiterpenes in the essential oil were 22.76%. This is the first report of the Agar-Sit and also the application of R. vinctus in agarwood induction. According to the results, when the combination of Agar-Sit and R. vinctus is used agarwood can be induced with high yield and good quality.

Porostereum sp., Associated with Saffron (Crocus sativus L.), is a Latent Pathogen Capable of Producing Phytotoxic Chlorinated Aromatic Compounds.

Saffron (Crocus sativus L.) is one of the most expensive spices in the world due to its medicinal and aromatic value. However, saffron production is severely affected by the corm rot disease throughout the saffron producing countries. In this study, we report a basidiomycetous latent pathogen of saffron, designated as CSE26, capable of producing phytotoxic compounds. CSE26 is a highly odorous basidiomycete with monomitic hyphal system. Molecular phylogeny of ITS and 28S ribosomal gene sequence of CSE26 assigned it as Porostereum spadiceum. It was found to produce corm rot in C. sativus under in vivo and field conditions, with a disease severity index of 0.7 and 0.5, respectively. CSE26 was found to produce chlorinated aromatic compounds (CAMs) having phytotoxic activity against Arabidopsis plants. Therefore, these compounds may be acting as pathogenic determinants of CSE26. However, there is a need to study the level of production of these CAMs by this fungus in the natural environment and their effects on plant health.

Wood decay fungus Flavodon ambrosius (Basidiomycota: Polyporales) is widely farmed by two genera of ambrosia beetles.

The ambrosia fungus Flavodon ambrosius is the primary nutritional mutualist of ambrosia beetles Ambrosiodmus and Ambrosiophilus in North America. F. ambrosius is the only known ambrosial basidiomycete, unique in its efficient lignocellulose degradation. F. ambrosius is associated with both native American beetle species and species introduced from Asia. It remains unknown whether F. ambrosius is strictly a North American fungus, or whether it is also associated with these ambrosia beetle genera on other continents. We isolated fungi from the mycangia and galleries of ambrosia beetles Ambrosiodmus rubricollis, Ambrosiodmus minor, Ambrosiophilus atratus, and Ambrosiophilus subnepotulus in China, South Korea, and Vietnam. Phylogenetic analyses suggest that all Asian and North American isolates represent a single haplotype. These results confirm Flavodon ambrosius as the exclusive mutualistic fungus of multiple Ambrosiodmus and Ambrosiophilus beetle species around the world, making it the most widespread known ambrosia fungus species, both geographically and in terms of the number of beetle species. The Flavodon-beetle symbiosis appears to employ an unusually strict mechanism for maintaining fidelity, compared to the symbioses of the related Xyleborini beetles, which mostly vector more dynamic fungal communities.

New Records of Hyphoderma (Meruliaceae, Polyporales) for India.

An account of eight species of genus Hyphoderma (H. clavatum, H. definitum, H. echinocystis, H. litschaueri, H. nemorale, H. subpraetermissum, H. tibia, and H. transiens) is presented, which is based on collections made from Uttarakhand state during 2009-2014. All these species are cited and fully described for the first time from India.

Extensive characterization of the new genus Rickiopora (Polyporales).

Ceriporiopsis latemarginata and Antrodiella angulatopora are two Neotropical polypores that are very similar morphologically and are characterized by effused-reflexed basidiomes, large angular pores, small ellipsoid to ovoid basidiospores and a monomitic hyphal system with presence of thick-walled hyphae with infrequent clamps. In order to verify the evolutionary relations of C. latemarginata and to find its taxonomic placement, morphological, biological, and phylogenetic studies were carried out, as well as comparisons with Antrodiella s.s. and Ceriporiopsis s.s. Mono- and poly-sporic cultures were obtained and used for mating system and nuclear behaviour studies, as well as for DNA sequence analyses of the internal transcribed spacer and large subunit of ribosomal RNA gene, translation elongation factor 1-alpha gene and RNA polymerase II second largest subunit-like gene. Type specimens of C. latemarginata and A. angulatopora were morphologically examined and their synonymy was confirmed. The results obtained support the description of a new polypore genus Rickiopora in the residual polyporoid clade characterized by a bipolar mating system, an astatocoenocytic nuclear behaviour, and a monomitic hyphal system that display a metachromatic reaction in cresyl blue.

New Fungus-Insect Symbiosis: Culturing, Molecular, and Histological Methods Determine Saprophytic Polyporales Mutualists of Ambrosiodmus Ambrosia Beetles.

Ambrosia symbiosis is an obligate, farming-like mutualism between wood-boring beetles and fungi. It evolved at least 11 times and includes many notorious invasive pests. All ambrosia beetles studied to date cultivate ascomycotan fungi: early colonizers of recently killed trees with poor wood digestion. Beetles in the widespread genus Ambrosiodmus, however, colonize decayed wood. We characterized the mycosymbionts of three Ambrosiodmus species using quantitative culturing, high-throughput metabarcoding, and histology. We determined the fungi to be within the Polyporales, closely related to Flavodon flavus. Culture-independent sequencing of Ambrosiodmus minor mycangia revealed a single operational taxonomic unit identical to the sequences from the cultured Flavodon. Histological sectioning confirmed that Ambrosiodmus possessed preoral mycangia containing dimitic hyphae similar to cultured F. cf. flavus. The Ambrosiodmus-Flavodon symbiosis is unique in several aspects: it is the first reported association between an ambrosia beetle and a basidiomycotan fungus; the mycosymbiont grows as hyphae in the mycangia, not as budding pseudo-mycelium; and the mycosymbiont is a white-rot saprophyte rather than an early colonizer: a previously undocumented wood borer niche. Few fungi are capable of turning rotten wood into complete animal nutrition. Several thousand beetle-fungus symbioses remain unstudied and promise unknown and unexpected mycological diversity and enzymatic innovations.

Genetic resources and mycelial characteristics of several medicinal polypore mushrooms (Polyporales, Basidiomycetes).

Mycelial characteristics of dikaryotic collections of 6 medicinal polypore mushrooms (Fomes fomentarius, Fomitopsis pinicola, Ganoderma adspersum, G. applanatum, G. lucidum, and G. resinaceum) with different geographical origins (Armenia, China, France, Iran, Italy, and Russia) were screened. A total of 42 polypore collections were molecularly identified by sequencing the internal transcribed spacer region of the ribosomal RNA genes' cluster, and a phylogenetic tree was constructed. Morphological characteristics of 37 cultures were observed on agar media (malt extract agar, potato dextrose agar) at different temperatures (25, 30, 35, and 38°C) at a pH of 6.0. Colony morphology, pigmentation of mycelium and agar, mycelial growth rate, in vitro teleomorph formation, and other macromorphological characteristics were thoroughly described and illustrated. Micromorphological features of mycelia, such as different hyphal structures, clamp cells, presence and type of asexual sporulation, chlamydospores, and others were observed. The taxonomic significance of the mycelial characteristics revealed was estimated. The obtained results will assist further biotechnological cultivation of medicinal polypore mushrooms to develop novel health care biotechnological products.

Possible Tyromyces fissilis (Basidiomycota, Polyporales) co-infection in a lung transplant recipient.

Invasive fungal diseases are severe complication of the lung transplant patients' follow-up as they are increasing the risk of rejection. We report a patient who developed possible Tyromyces fissilis co-infection during graft rejection episode 2 years after bilateral lung transplantation for cystic fibrosis. The fungus was detected using conventional culture methods as a filamentous basidiomycete and further placed to T. fissilis species based on internal transcribed spacer (ITS) rDNA sequences. The patient was treated according to the susceptibility testing results by voriconazole in combination with the anti-rejection therapy and recovered completely within few weeks. This is, to our knowledge, the first published case report of T. fissilis as a possible causative agent of an infection/rejection episode in a lung transplant recipient.

Molecular phylogeny of Rigidoporus microporus isolates associated with white rot disease of rubber trees (Hevea brasiliensis).

Rigidoporus microporus (Polyporales, Basidiomycota) syn. Rigidoporus lignosus is the most destructive root pathogen of rubber plantations distributed in tropical and sub-tropical regions. Our primary objective was to characterize Nigerian isolates from rubber tree and compare them with other West African, Southeast Asian and American isolates. To characterize the 20 isolates from Nigeria, we used sequence data of the nuclear ribosomal DNA ITS and LSU, ß-tubulin and translation elongation factor 1-α (tef1) gene sequences. Altogether, 40 isolates of R. microporus were included in the analyses. Isolates from Africa, Asia and South/Central America formed three distinctive clades corresponding to at least three species. No phylogeographic pattern was detected among R. microporus collected from West and Central African rubber plantations suggesting continuous gene flow among these populations. Our molecular phylogenetic analysis suggests the presence of two distinctive species associated with the white rot disease. Phylogenetic analyses placed R. microporus in the Hymenochaetales in the vicinity of Oxyporus. This is the first study to characterize R. microporus isolates from Nigeria through molecular phylogenetic techniques, and also the first to compare isolates from rubber plantations in Africa and Asia.

Phylogenetic relationships of Korean Sparassis latifolia based on morphological and ITS rDNA characteristics.

Recent studies based on morphological characteristics and molecular analyses have revealed that the characteristics of Sparassis crispa from Asia are not concordant with those of collections from Europe and North America. Consequently, the Asian isolate was redefined as Sparassis latifolia. This study is the first report of Sparassis latifolia collected in Korea. The taxonomic relationships and replacement of Sparassis species were inferred from a comparison of the morphological characteristics and by molecular sequence analysis of the internal transcribed spacer (ITS) rDNA regions. In particular, this study focused on the phylogenetic relationships inferred from the biogeographical distribution of isolates within the genus Sparassis.

The phylogenetic position of Postia s.l. (Polyporales, Basidiomycota) from Patagonia, Argentina.

We investigated the phylogenetic relationships of Postia species from Patagonia with rDNA ITS and LSU sequences, together with morphological, cultural and biological features. All species in the genus were included in a "Postia clade" irrespective of whether their spores were thin- or thick-walled. This clade is characterized by tetrapolar mating, a normal nuclear behavior, metachromatic generative hyphae and absence of fiber hyphae in culture. One subclade merged the austral taxa P. pelliculosa and P. punctata, but otherwise no distinct relationships were found regarding spore shape, spore wall thickness and geographical distribution of taxa. The austral P. venata and the endemic P. carbophila, with thin-walled basidiospores, occupied variable positions in both analyses. Postia caesia from Patagonia grouped within the P. caesia species complex but on a separate branch. In contrast, P. rennyi and P. balsamea from Patagonia corresponded well with strains from other geographic areas. The two austral species in Ryvardenia, R. cretacea and R. campyla, characterized by non-metachromatic hyphae, bipolar mating and an astatocoenocytic nuclear behavior, formed an independent subclade among the dimitic genera of the "Antrodia clade", far from other Postia taxa within which they had been placed previously, supporting their inclusion in a genus of their own. Postia carbophila grouped with other Postia species and not with Postia (Rhodonia) placenta as suggested previously on the basis of morphological comparisons. Instead, the latter species grouped with taxa in the dimitic genus Amyloporia with which it shares heterocytic nuclear behavior. A separation between specimens of Postia pelliculosa and Ryvardenia cretacea from either side of the Pacific (i.e. SE Australia/New Zealand and S Argentina/S Chile) suggests they could be considered different at the species level from a phylogenetic point of view.

A new species of Hyphoderma (Meruliaceae, Polyporales) and its discrimination from closely related taxa.

Thirty-five corticioid collections from the Canary Islands and Azores Archipelago were examined morphologically and subjected to molecular phylogenetic analysis. These specimens, almost all collected on endemic and/or xerophilic vegetation, were similar in morphological and ecological characteristics to Hypochnicium prosopidis from the Sonoran Desert (Arizona, USA) and Hyphoderma amoenum. Thirty-seven new ITS nrDNA sequences from these specimens, including the nomenclatural type of the above-mentioned species, were obtained and aligned with homologous sequences from GenBank. These collections were distributed in two strongly supported monophyletic clades. However, similar patterns of morphological variability shared by specimens included in both clades and their differences with related species suggest that they should be described as a single new species. Therefore Hyphoderma macaronesicum is proposed. Studies will be required to test, in a more robust multilocus genealogical framework, whether these populations constitute two cryptic species or whether they are the same taxon. The position of Hypochnicium prosopidis in the resolved tree and its morphological characters suggest that it should be included in Hyphoderma and the new combination Hyphoderma prosopidis is proposed.

Wood-inhabiting fungi in southern China 5. New species of Theleporus and Grammothele (Polyporales, Basidiomycota).

During the examination of specimens of Theleporus and Grammothele (Polyporales, Basidiomycota) from tropical China, three new species, Theleporus membranaceus, T. minisporus and Grammothele denticulata, were identified based on both morphological and phylogenetic analyses. They are described and illustrated. T. membranaceus is characterized by its extremely thin basidiocarps (0.12 mm), small pores (7-10 per mm) and ellipsoid to broadly ellipsoid basidiospores. T. minisporus has the smallest basidiospores among the species in the genus. Grammothele denticulata is distinguished in the genus by gray pores, continuous hymenia over dissepiment edge and cylindrical basidiospores with tapering apex. Two annotated identification keys are provided for species thus far accepted in Theleporus and Grammothele. The phylogenetic relationships of Theleporus and Grammothele were inferred based on nITS sequences and are briefly discussed. The molecular evidence showed that Theleporus and Grammothele belong to the core polyporoid clade.

Differential regulation of laccase gene expression in Coriolopsis rigida LPSC No. 232.

Two laccase isoenzyme genes (lcc2 and lcc3) from the white-rot fungus Coriolopsis rigida were cloned, and together with the previously described lcc1, their transcript levels were analysed by Quantitative RT-PCR in order to study their expression patterns under a range of putative inducers (Cu(2+), Mn(2+), Fe(3+), 2,6-dimethoxy-1,4-benzoquinone, H(2)O(2,) caffeine, amphotericin B and syringic acid). The highest induction was observed for lcc1 in presence of copper, and thus, a kinetic study was performed to analyze its effect on temporary lcc1 gene expression. Our results showed that upregulation due to copper was linked to growth stage, being highest during the trophophase and decreasing during the idiophase. Amphotericin B increased levels of transcripts of lcc1 and lcc2, syringic acid upregulated lcc1 and lcc3 and 2,6-dimethoxy-1,4-benzoquinone induced lcc2 and lcc3. Possible reasons for why laccase genes from C. rigida are differentially regulated at the transcriptional level are discussed.

Bondarzewia podocarpi, a new and remarkable polypore from tropical China.

Bondarzewia podocarpi is described as new and is distinguished from other species in the genus by regular pores, presence of cystidioles, thin generative hyphae and short basidia. It was found in tropical China and grows on living trees of Podocarpus. Parsimony analyses were applied to an ITS dataset. The results suggest that Bondarzewia is monophyletic with two well supported internal clades. One clade includes B. podocarpi and B. guaitecasensis, while the other includes B. berkeleyi and B. montana. Based on studied materials of Bondarzewia from the world, an identification key to the species of the genus is provided.

Two new species of Hyphodontia from China.

Hyphodontia septocystidiata and Hyphodontia sinensis are reported as new to science. Hyphodontia septocystidiata has grandinioid to odontioid hymenophore, encrusted cystidia, septocystidia and capitate cystidia and broadly ellipsoid to subglobose thick-walled basidiospores. Hyphodontia sinensis has odontioid hymenophore, leptocystidia and capitate cystidioles and broadly ellipsoid, thin- to slightly thick-walled basidiospores. Descriptions and line drawings are provided for these two new species.