Application of CRISPR/Cas9 Tools for Genome Editing in the White-Rot Fungus Dichomitus squalens.

Dichomitus squalens is an emerging reference species that can be used to investigate white-rot fungal plant biomass degradation, as it has flexible physiology to utilize different types of biomass as sources of carbon and energy. Recent comparative (post-) genomic studies on D. squalens resulted in an increasingly detailed knowledge of the genes and enzymes involved in the lignocellulose breakdown in this fungus and showed a complex transcriptional response in the presence of lignocellulose-derived compounds. To fully utilize this increasing amount of data, efficient and reliable genetic manipulation tools are needed, e.g., to characterize the function of certain proteins in vivo and facilitate the construction of strains with enhanced lignocellulolytic capabilities. However, precise genome alterations are often very difficult in wild-type basidiomycetes partially due to extremely low frequencies of homology directed recombination (HDR) and limited availability of selectable markers. To overcome these obstacles, we assessed various Cas9-single guide RNA (sgRNA) ribonucleoprotein (RNP) -based strategies for selectable homology and non-homologous end joining (NHEJ) -based gene editing in D. squalens. We also showed an induction of HDR-based genetic modifications by using single-stranded oligodeoxynucleotides (ssODNs) in a basidiomycete fungus for the first time. This paper provides directions for the application of targeted CRISPR/Cas9-based genome editing in D. squalens and other wild-type (basidiomycete) fungi.

Cost-effective optimized scleroglucan production by Sclerotium rolfsii ATCC 201126 at bioreactor scale. A quantity-quality assessment.

Exopolysaccharide (EPS) secretion by Sclerotium rolfsii ATCC 201126 in submerged cultures, already identified as high-osmolarity responsive, was assessed by reducing C-source without compromising EPS yields. A designed medium with 80 g sucrose L-1 (MOPT80) was tested at 3 L-bioreactor scale at different temperature, agitation, aeration and pH (uncontrolled vs. controlled) values. Optimal operative conditions (200 rpm, 28 °C, 0.5 vvm and initial pH -pHi- 4.5) were validated, as well as the possibility to work at pHi 5.5 to reduce biomass production. Purified EPSs produced in MOPT80 at optimal and other valid operative conditions exhibited refined grade (<1 % proteins and ash, 3-4 % reducing sugars, 87-99 % total sugars). EPS purity, MW and rheological parameters led to discourage pH controlled at 4.5. Relatively constant MW (6-8 × 106 Da) and outstanding viscosifying ability were found. Polyphasic EPS analysis (titre, purity, macromolecular features and rheological fitness) would support to properly select production conditions.

Differences in aluminum tolerance and immobilization between two indigenous ectomycorrhizal fungi Lactarius deliciosus and Pisolithus tinctorius from Southwest China's forest stands.

Aluminum (Al) toxicity severely decreases plant growth and productivity in acidic soil globally. Ectomycorrhizal (ECM) fungi can promote host plant's Al-tolerance by acting as a physical barrier or bio-filter. However, little information is available on the role of ECM fungus on Al immobilization with respect to Al-tolerance. This present study aimed to screen a promising indigenous ECM fungus with high Al-tolerance and to understand its role in Al immobilization related to Al-tolerance. Two ECM fungal strains (Lactarius deliciosus 2 and Pisolithus tinctorius 715) isolated from forest stands in Southwest China were cultured in vitro with 0.0, 1.0 or 2.0 mM Al addition for 21 days to compare their Al accumulation and Al-tolerance. Meanwhile, fungal mycelia were incubated in 0.037 mM Al3+ solutions, and then Al3+ concentrations in the solution were determined at time 2, 5, 10, 20, 40, 60, 120, 180, and 240 min, and the Al3+ immobilization characteristics were evaluated using the pseudo-first order, pseudo-second order and intraparticle diffusion models. Results showed that 1.0 or 2.0 mM Al3+ addition significantly increased fungal biomass production by 23% or 41% in L. deliciosus 2, not in P. tinctorius 715. Fungal Al3+ concentrations in L. deliciosus 2 and P. tinctorius 715 were significantly increased by 293% and 103% under 2.0 mM than under 1.0 mM Al3+ addition. The pH values in the culture solution were significantly decreased by 0.43 after 21 d fungus growth but no changes between these two fungi under the same Al3+ addition. Fungal Al3+ immobilization showed a three-stage trend with initially a rapid rate followed a relatively slower rate until reaching equilibrium. The pseudo-second order model was the best (R2 = 0.98 and 0.99 for L. deliciosus 2 and P. tinctorius 715) to fit the experimentally observed data among the three models. Compared to P. tinctorius 715, L. deliciosus 2 also had greater intercept value, cation exchange capacity (CEC), and extracellular Al3+ proportion in fungal mycelia. Additionally, bio-concentration on Al3+, active site numbers for Al3+, boundary layer thickness, CEC, and immobilization on the cell wall in fungal mycelia were involved in ECM fungal Al-tolerance. These results show that both ECM fungi are Al-tolerant while L. deliciosus 2 is a promising indigenous ECM isolate with higher Al-tolerance in Southwest China, and they can be hence applied to the afforestation and ecological restoration in acidic soil.

Degradation of the Organochlorinated Herbicide Diuron by Rainforest Basidiomycetes.

The main organochlorinated compounds used on agricultural crops are often recalcitrant, affecting nontarget organisms and contaminating rivers or groundwater. Diuron (N-(3,4-dichlorophenyl)-N',N'-dimethylurea) is a chlorinated herbicide widely used in sugarcane plantations. Here, we evaluated the ability of 13 basidiomycete strains of growing in a contaminated culture medium and degrading the xenobiotic. Dissipation rates in culture medium with initial 25 mg/L of diuron ranged from 7.3 to 96.8%, being Pluteus cubensis SXS 320 the most efficient strain, leaving no detectable residues after diuron metabolism. Pycnoporus sanguineus MCA 16 removed 56% of diuron after 40 days of cultivation, producing three metabolites more polar than parental herbicide, two of them identified as being DCPU and DCPMU. Despite of the strong inductive effect of diuron upon laccase synthesis and secretion, the application of crude enzymatic extracts of P. sanguineus did not catalyzed the breakdown of the herbicide in vitro, indicating that diuron biodegradation was not related to this oxidative enzyme.

Biological activities of melanin pigment extracted from Bombyx mori gut-associated yeast Cryptococcus rajasthanensis KY627764.

Melanin pigment has been produced and extracted from a wide variety of living forms ranging from microorganisms to higher organisms. Owing to the therapeutic nature of the pigment, various microbial populations have been explored for its production. Hence, we isolated a melanin producing yeast from the insect Bombyx mori gut microflora and identified it as Cryptococcus rajasthanensis based on the molecular characterization. The isolated yeast produced enhanced melanin pigment when cultured in the minimal L-tyrosine broth as compared to the Saboraud medium. The pigment was extracted and characterized as melanin based on UV-Visible spectroscopy, FTIR (Fourier-transform infrared) spectroscopy and 1H NMR (Nuclear magnetic resonance). The melanin pigment was evaluated as a potent bioactive molecule with bioactivity like antimicrobial, antioxidant, anti-inflammatory, and anticancer activity that describes the therapeutic nature of the extracted melanin pigment. Distinct from the biologically active role the melanin pigment isolated from the yeast, the Cryptococcus extract also exhibited killer toxin activity against the pathogenic yeast Candida albicans.

Relationship between the honeydew of mealy bugs and the growth of Phlebopus portentosus.

BACKGROUND: Phlebopus portentosus and mealy bugs form a fungus-insect gall on the roots of host plants. The fungus and mealy bugs benefit mutually through the gall, which is the key link in the nutritional mechanism of P. portentosus. The cavity of the fungus-insect gall provides an ideal shelter for mealy bugs survival and reproduction, but how does P. portentosus benefit from this symbiotic relationship? METHODOLOGY AND RESULTS: Anatomical examination of fungus-insect galls revealed that one or more mealy bugs of different generations were living inside the galls. The mealy bug's mouthpart could penetrate through the mycelium layer of the inside of the gall and suck plant juice from the host plant root. Mealy bugs excreted honeydew inside or outside the galls. The results of both honeydew agar medium and quartz tests showed that the honeydew can attract and promote the mycelial growth of P. portentosus. A test of the relationship between the honeydew and the formation of the fungus-insect gall showed that honeydew promoted gall formation. CONCLUSIONS: All experimental results in this study show that the honeydew secreted by mealy bugs can attract and promote the mycelial growth of P. portentosus, forming a fungus-insect gall, because mealy bugs' honeydew is rich in amino acids and sugars.

Combined Effect of Light and Nutrients on the Micromorphology of the White rot Fungus Cerrena Unicolor.

Light influences developmental pathways in fungi. Recent transcriptomic and biochemical analyses have demonstrated that light influences the metabolism of a white-rot basidiomycete Cerrena unicolor. However, the expression profile of genes involved in the growth and development, or micromorphological observations of the mycelium in response to variable lighting and culturing media, have not performed. We aim to reveal the effect of light and nutrients on C. unicolor growth and a potential relationship between the culture medium and lighting conditions on fungus micromorphological structures. Confocal laser scanning microscopy and scanning electron microscopy were employed for morphological observations of C. unicolor mycelium cultivated in red, blue, green, and white light and darkness on mineral and sawdust media. A comprehensive analysis of C. unicolor differentially expressed genes (DEGs) was employed to find global changes in the expression profiles of genes putatively involved in light-dependent morphogenesis. Both light and nutrients influenced C. unicolor growth and development. Considerable differences in the micromorphology of the mycelia were found, which were partially reflected in the functional groups of DEGs observed in the fungus transcriptomes. A complex cross-interaction of nutritional and environmental signals on C. unicolor growth and morphology was suggested. The results are a promising starting point for further investigations of fungus photobiology.

Basidioradulum mayi and B. tasmanicum spp. nov. (Hymenochaetales, Basidiomycota) from both sides of Bass Strait, Australia.

Basidioradulum was morphologically considered to be a synonym of Xylodon. Here, its independence within Hymenochaetales is confirmed from a phylogenetic perspective. Basidioradulum radula, the generic type, is widely distributed in Northern Hemisphere. Two Southern Hemisphere species close to B. radula are newly described as B. mayi and B. tasmanicum, respectively, from Victoria and Tasmania, Australia. Basidioradulum mayi differs from B. radula by lack of cystidia. Moreover, the hymenial surface of B. radula is normally much more strongly hydnoid than that of B. mayi. Basidioradulum tasmanicum is distinct from B. radula and B. mayi by having capitate cystidia, ellipsoid to subglobose basidiospores, and crystal-covered hyphae. Although morphologically distinct, the two new species isolated by Bass Strait have an almost identical ITS region, and could not be differentiated by nLSU- and ITS-based phylogenetic analyses. This case reminds us that basing phylogeny simply on the ITS as a barcode region may underestimate fungal species diversity.

Bio-based Preservative using Methyl-ß-cyclodextrin-Essential Oil Complexes for Wood Protection.

Developing natural preservatives for wood protection is of great interest in sustainable construction and green building industries. This study was designed to determine the feasibility of using methyl-ß-cyclodextrin-essential oils (MßCD-EOs) complexes as potential bio-based preservatives for wood protection. Four essential oils (EOs) with proven antifungal properties, eugenol (EG), trans-cinnamaldehyde (CN), thymol (TM) and carvacrol (CV), were complexed with methyl-ß-cyclodextrin (MßCD) by a co-precipitation method. The inclusion of EOs in MßCD and the corresponding inclusion yield of the MßCD-EOs complexes were determined by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and ultraviolet-visible spectroscopy (UV/Vis), respectively. The maximum inclusion yields in MßCD-EG, MßCD-CN, MßCD-CV, and MßCD-TM were estimated to be almost 100%. MßCD-EOs complexes were impregnated into southern pine wood blocks and exposed to two brown rot fungi, Gloeophyllum trabeum and Postia placenta, following procedures described in AWPA Standard E22 and E10. The penetration of MßCD-EOs complexes in wood was confirmed by fluorescence microscopic analysis after the selective dyeing of EOs encapsulated in MßCD. In comparison to the control wood samples, MßCD-EOs complexes treated wood exhibited a significant reduction in the mass loss from 16-36% to 2-18%, accompanied by improvement in radial compression strength loss from 81-92% to 29-67% after four-week fungi exposure.

Molecular Cloning, Characterisation, and Heterologous Expression of Farnesyl Diphosphate Synthase from Sanghuangporus baumii.

A farnesyl diphosphate synthase (FPS) cDNA and promoter region was cloned from Sanghuangporus baumii. The gene contains a 150-bp 5'-untranslated region (UTR), a 154-bp 3'-UTR, and a 1062-bp open reading frame (ORF) encoding a 354 amino acid polypeptide. The FPS-DNA includes three exons (nucleotides 1 -123, 184-321, and 505-1305) and two introns (nucleotides 124-183 and 322-504). The FPS protein has a molecular weight of 40.73 kDa, it is hydrophilic with a theoretical isoelectric point of 5.13, and the secondary and three-dimensional structure were analysed. There is a transcription start site at nucleotides 1318-1368 of the promoter, which includes typical eukaryotic promoter elements (TATA Box, CAAT Box, ARBE, AT-rich element, G-box, MBS, Sp1, LTR). FPS was expressed in Escherichia coli BL21, and the recombinant protein (63.41 kDa) was subjected to dodecyl sulphate, sodium salt-polyacrylamide gel electrophoresis (SDS-PAGE). FPS transcription was measured during different developmental stages, and expression in 11 and 13 days mycelia was upregulated 49.3-fold and 125.4-fold, respectively, compared with 9 days mycelia controls. Through analysing, S. baumii triterpenoid content was correlated with the transcription level of FPS during different development stages, and the triterpenoid content peaked at day 15 (7.21 mg/g).

Apiotrichum veenhuisii isolated from a pediatric patient with acute myeloid leukemia: The first case in humans.

This is the first report of the yeast Apiotrichum veenhuisii (formerly Trichosporon veenhuisii) causing disease in humans; its virulence and in vitro behavior against antifungals were also studied. The sample was isolated from biopsy fragments of disseminated lesions on the skin of a pediatric patient with acute myeloid leukemia. The studied virulence factors evidenced that the strain tested negative for secretion of the enzymes proteinase, phospholipase, and hemolysin. The isolate was characterized as low biofilm producer. Except for amphotericin B and voriconazole, the sample presented high minimum inhibitory concentration values against azole and echinocandins.

Comparative secretome analysis of different smut fungi and identification of plant cell death-inducing secreted proteins from Tilletia horrida.

BACKGROUND: Tilletia horrida is a basidiomycete fungus that causes rice kernel smut, one of the most important rice diseases in hybrid rice growing areas worldwide. However, little is known about its mechanisms of pathogenicity. We previously reported the genome of T. horrida, and 597 genes that encoded secreted proteins were annotated. Among these were some important effector genes related to pathogenicity. RESULTS: A secretome analysis suggested that five Tilletia fungi shared more gene families than were found in other smuts, and there was high conservation between them. Furthermore, we screened 597 secreted proteins from the T. horrida genome, some of which induced expression in host-pathogen interaction processes. Through transient expression, we demonstrated that two putative effectors could induce necrosis phenotypes in Nicotiana benthamiana. These two encoded genes were up-regulated during early infection, and the encoded proteins were confirmed to be secreted using a yeast secretion system. For the putative effector gene smut_5844, a signal peptide was required to induce non-host cell death, whereas ribonuclease catalytic active sites were required for smut_2965. Moreover, both putative effectors could induce an immune response in N. benthamiana leaves. Interestingly, one of the identified potential host interactors of smut_5844 was laccase-10 protein (OsLAC10), which has been predicted to be involved in plant lignification and iron metabolism. CONCLUSIONS: Overall, this study identified two secreted proteins in T. horrida that induce cell death or are involved in defense machinery in non-host plants. This research provides a useful foundation for understanding the interaction between rice and T. horrida.

Selenium biofortification and its effect on multi-element change in Auricularia auricular.

Auricularia auricular could be useful as a candidate for human selenium supplementation. This study examined the effects of exogenous Se on the growth, yield, nutritive value, and mineral accumulation of A. auricular. Selenate or selenite (0.5-40.0 µg g-1) had no effect on mycelium morphology or the yield of fruiting bodies. In some cases, they affected the accumulation of inter-elements and significantly decreased the concentrations of copper, iron, and chromium in the Se-enriched fruiting bodies compared to that with control treatments. The polysaccharide (116.5-131.6 µg g-1) and protein (105.2-113.4 µg g-1) content in Se-enriched fruiting bodies were not significantly different from those observed in the controls (polysaccharide, 114.1 µg g-1; protein, 105.6 µg g-1). Thus, A. auricular can absorb inorganic Se from the substrate and convert it to organic Se compounds (selenocystine (≥4.1%), selenomethionine (≥91.9%), and Se-methylselenocysteine (≥2.3%)).

Production, structural characterization, and antiproliferative activity of exopolysaccharide produced by Scleroderma areolatum Ehrenb with different carbon source.

The effects of different three carbon sources, that is, glucose, fructose, and sucrose, on production, molecular properties and antiproliferative activity of exopolysaccharide (EPS), were evaluated in the submerged culture of Scleroderma areolatum Ehrenb. Among carbon sources examined, the addition of sucrose maximizes the mycelia production, while fructose could maximize the EPS yield. Although the predominant carbohydrate compositions identified were gluconic acid and mannose, the monosaccharide composition of EPSs was also different significantly. FT-IR spectral analysis revealed there was no significant difference among the prominent characteristic groups in three EPSs. The molecular weight of EPSs was also affected by carbon source, being generally lower compared with that with glucose. However, all EPSs molecule existed as nearly globular shape form in aqueous solution. The variation of carbon sources also affected antiproliferative activity examined in vitro using cell proliferation assay. Fructose was optimal carbon source giving higher antiproliferative activity probably due to the relatively high contents of xylose in the EPS with low molecular weight.

Single Cell Oil Production from Undetoxified Arundo donax L. hydrolysate by Cutaneotrichosporon curvatus.

The use of low-cost substrates represents one key issue to make single cell oil production sustainable. Among low-input crops, Arundo donax L. is a perennial herbaceous rhizomatous grass containing both C5 and C6 carbohydrates. The scope of the present work was to investigate and optimize the production of lipids by the oleaginous yeast Cutaneotrichosporon curvatus from undetoxified lignocellulosic hydrolysates of steam-pretreated A. donax. The growth of C. curvatus was first optimized in synthetic media, similar in terms of sugar concentration to hydrolysates, by applying the response surface methodology (RSM) analysis. Then the bioconversion of undetoxified hydrolysates was investigated. A fed-batch process for the fermentation of A. donax hydrolysates was finally implemented in a 2-L bioreactor. Under optimized conditions, the total lipid content was 64% of the dry cell weight and the lipid yield was 63% of the theoretical. The fatty acid profile of C. curvatus triglycerides contained 27% palmitic acid, 33% oleic acid and 32% linoleic acid. These results proved the potential of lipid production from A. donax, which is particularly important for their consideration as substitutes for vegetable oils in many applications such as biodiesel or bioplastics.

Medicinal, Nutritional, and Cosmetic Values of Macrofungi Distributed in Mazandaran Province of Northern Iran (Review).

A total of 82 species and 2 variations of medicinal and edible macrofungi (Basidiomycota and Ascomy-cota) have been collected and identified in Mazandaran Province of Northern Iran. Among these species, 58 possess medicinal and culinary properties, whereas 21 possess only medicinal properties (antiinflammatory, antimicrobial, an-timutagenic, antioxidant, antitumor, antiplatelet, hypoglycemic, hypocholesterolemic, neurotropic, hypotensive, insec-ticidal, immunomodulating, mitogenic/regenerative, spasmolytic, etc.). No medicinal effect has yet been reported for 4 species (Cantharellus infundibuliformis, Mycena inclinata, Suillus collinitus, Xerocomus porosporus) and 2 variations (Pluteus cervinus var. albus, Russula cyanoxantha var. variata) of edible species. Among the listed species, 15 (such as Auricularia auricula-judae, Hericium erinaceus, Fomes fomentarius, Flammulina velutipes, Ganoderma lucidum, Lyophyllum decastes, Pleurotus ostreatus, Tremella mesenterica) are currently used for manufacturing organic cosmetic products. The well-known mushrooms with medicinal properties (e.g., Auricularia auricula-judae, Flammulina velutipes, Hericium erinaceus, Ganoderma lucidum, G. tsugae, G. adspersum, Trametes versicolor) and excellent edibility (e.g., Boletus edulis, Cantharellus cibarius, Morchella esculenta, Pleurotus ostreatus) are widely distributed in the studied area. The biological resources of macrofungi growing in Mazandaran Province of Northern Iran possess medicinal, nutritional, and cosmetic values and could be further used for biotechnological exploitation to develop mushroom-derived pharmaceuticals, nutriceuticals, nutraceuticals, and cosmeceuticals.

Aislamiento y producción de basidiomas de cepas nativas de Volvariella bombycina (Schaeff) Singer, utilizando desechos agrícolas / Isolation and production of basidiomes from native strains of Volvariella bombycina (Schaeff) Singer, using agro-industrial wastes

Volvariella bombycina es una especie de hongo saprófito comestible que crece en troncos de árboles muertos y sobre la cual existe abundante información acerca de su sabor y propiedades nutricionales y medicinales. En Guatemala esta especie se consume en diversas regiones templado-cálidas del país, pero a la fecha no existe ningún estudio sobre su cultivo a nivel local. La presente investigación se realizó con el fin de conocer el comportamiento de tres cepas guatemaltecas de V. bombycina cultivadas en desechos agroindustriales. Las cepas fueron aisladas de basidiomas recolectados en bosques cercanos a la Ciudad de Guatemala, utilizándose medio PDA para el aislamiento y temperatura de 30 ºC para incubación. En la producción de inóculo se calculó la tasa de extensión radial (RER) sobre granos de sorgo y trigo en cajas de Petri. Para la obtención de basidiomas se evaluaron seis sustratos de desechos agrícolas: pasto jaraguá, caña y olote de maíz, rastrojo de frijol, paja de trigo, paja de arroz y tronco de tonché (Ipomoea murucoides), en frascos de vidrio. Se encontró que no hubo diferencia estadística significativa en el análisis de la RER entre las cepas VNPNU-01 y AADM-01 (p > .05), pero sí con la cepa VNPNU-02 (p < .05). Se obtuvieron basidiomas de V. bombycina en los seis sustratos evaluados, siendo la cepa AADM-01 la más productiva. De acuerdo con este estudio, las tres cepas guatemaltecas de V. bombycina pueden ser utilizadas en la producción de basidiomas en desechos agrícolas, lo que permitiría obtener un alimento nutritivo y generar ingresos a quienes deseen cultivar este hongo.

Volvariella bombycina is a species of edible saprophyte fungus that grows on logs of dead trees and about which there is information about its taste and nutritional and medicinal properties. In Guatemala this species is consumed in several warm regions of the country, but there is still no study on its cultivation locally. The present investigation was carried out in order to know the behavior of three Guatemalan strains of V. bombycina when cultivated in agricultural wastes. The strains were isolated from basidiomata collected in forests close to Guatemala City, using PDA medium and a temperature of 30ºC for incubation. In the inoculum production, the radial extension rate (RER) was calculated using sorghum and wheat grains in Petri dishes. For obtaining basidiomata, six agricultural wastes substrates were analyzed: grazing grass, cane and corn cob, stubble of bean, wheat straw, rice straw and tonché trunk (Ipomoea murucoides), in glass jars. There was found no significant difference in the statistical analysis of the RER with the VNPNU-01 and AADM-01 strains (p > .05), but it was significant with the VNPNU-02 strain (p > .05). Basidiomata of V. bombycina were obtained in the six substrates evaluated, being the AADM-01 strain the most productive. According to this evaluation, the three Guatemalan strains of V. bombycina can be used in the production of basidiomas using agricultural wastes, what would allow the production of a nutritious food and generate another income for those who want to grow this mushroom.

A global view of Gyroporus: molecular phylogenetics, diversity patterns, and new species.

Gyroporus (Gyroporaceae, Boletales) is a highly diverse genus of poroid ectomycorrhizal mushrooms with a nearly worldwide distribution. Previous attempts to unravel the diversity within this genus proved difficult due to the presence of semicryptic species and ambiguous results from analysis of ribosomal RNA markers. In this study, we employ a combined morphotaxonomic and phylogenetic approach to delimit species and elucidate geographic and evolutionary patterns in Gyroporus. For phylogenetic analyses, the protein-coding genes atp6 (mitochondrial adenosine triphosphate [ATP] synthase subunit 6) and rpb2 (nuclear second largest subunit of RNA polymerase II) were selected based on their utility in studies of Boletales. We infer several distinct clades, most notably one corresponding to G. castaneus as a speciose Northern Hemisphere group, another unifying G. cyanescens and like entities, and a third group unifying G. longicystidiatus and a New World sister species. Also notable is the recovery of a sister relationship between the cyanescens and longicystidiatus clades. We formally describe five new species of Gyroporus, outline a number of provisional species, and briefly discuss distributional patterns. This study provides an important scaffold for future work on this well-known but poorly understood genus of fungi.

Taxonomy and phylogeny of Pyrrhoderma: a redefinition, the segregation of Fulvoderma, gen. nov., and identifying four new species.

Previously, Pyrrhoderma accommodated two polypore species, P. adamantinum and P. scaurum; however, phylogenetic studies indicated that these two species were not congeneric within the Hymenochaetaceae and that P. adamantinum formed a clade with Phellinidium noxium. To resolve the relationships among the two species of Pyrrhoderma and other related taxa, specimens from China, Costa Rica, Singapore, and Thailand were studied from both morphological and phylogenetic perspectives. A new genus, Fulvoderma, is erected to accommodate F. scaurum comb. nov., and a new species, F. australe (the generic type). Pyrrhoderma is delimited to include the generic type, P. sendaiense (a later synonym of P. adamantinum); two new combinations, P. lamaënse comb. nov., and P. noxium comb. nov.; and three new species, P. hainanense, P. thailandicum, and P. yunnanense. In addition, an undescribed lineage including several specimens from subtropical and tropical forests in China, Costa Rica, Singapore, and Thailand also nested within the Pyrrhoderma clade. However, as the voucher specimens are sterile or almost so, they are not described. The concept of Pyrrhoderma was emended to also accommodate species bearing resupinate, effuse-reflexed basidiocarps, hymenial or hyphoid setae, and non-subglobose basidiospores. Keys to Fulvoderma and Pyrrhoderma are provided.

Comprehensive analysis of a metabolic model for lipid production in Rhodosporidium toruloides.

The yeast Rhodosporidium toruloides has been extensively studied for its application in biolipid production. The knowledge of its metabolism capabilities and the application of constraint-based flux analysis methodology provide useful information for process prediction and optimization. The accuracy of the resulting predictions is highly dependent on metabolic models. A metabolic reconstruction for R. toruloides metabolism has been recently published. On the basis of this model, we developed a curated version that unblocks the central nitrogen metabolism and, in addition, completes charge and mass balances in some reactions neglected in the former model. Then, a comprehensive analysis of network capability was performed with the curated model and compared with the published metabolic reconstruction. The flux distribution obtained by lipid optimization with flux balance analysis was able to replicate the internal biochemical changes that lead to lipogenesis in oleaginous microorganisms. These results motivate the development of a genome-scale model for complete elucidation of R. toruloides metabolism.