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ABSTRACTLeccinum scabrum (Bull.) Gray (Boletaceae) is an edible mycorrhizal species with potential application interest due to its food and medicinal properties. A field investigation carried out during summer in the Białowieża Primeval Forest, located along the border between Belarus and Poland allowed to collect samples for chemical composition analysis and antibacterial activity evaluation. Mushroom extracts were prepared with microwave-assisted as well as ultrasound-assisted extraction techniques (UAE and MAE). The analysis of a dry sample of L. scabrum showed a significant content of vitamins and minerals and also a remarkable content of carbohydrates, protein, dietary fibre, total sugars, total free amino acids, and polyunsaturated fatty acids. The antibacterial activity of L. scabrum aqueous extracts showed inhibitory activity against all tested bacteria. In general, MAE extract exhibits a higher inhibition activity against Listeria monocytogenes ATCC 19114. As regards the Minimum Inhibitory Concentration (MIC) values, the high antibacterial activity of MAE extract was detected for L. monocytogenes ATCC 19114 and Escherichia coli ATCC 25922. Regarding UAE, high antibacterial activity was detected for Salmonella enterica ATCC 13076 and L. monocytogenes ATCC 19114. Based on data hereby reported, L. scabrum is a culinary-medicinal mushroom with a promising potential use as a high-quality food and nutraceutical mycological resource.

Mushrooms, a diverse group of fungi, has captivated human interest for centuries due to their intriguing characteristics and versatile application. The nutritional profile, coupled with unique organoleptic qualities, renders mushrooms not only a culinary delight, but also a valuable component of a balanced diet. In addition, mushrooms have gained recognition for their medicinal properties due to certain compounds found within them that exhibit promising pharmacological activities. Among the edible mushrooms, the Oyster mushroom (Pleurotus spp.) is commercially important in the global mushroom market and is widely cultivated and consumed in various parts of the world. Beyond the culinary appeal and medicinal properties of Pleurotus species, their cultivation holds an important environmental significance. These mushrooms are efficient decomposers of lignocellulosic biomass, contributing to agricultural and forestry waste recycling. Many researchers have demonstrated how their cultivation can serve as a sustainable practice, reducing the environmental impact of organic waste while generating a valuable food resource. In Sri Lanka, oyster mushroom cultivation is practiced on a small scale as a self-employed cottage industry. Four main species of oyster mushrooms, namely P. djamor, P. eous, P. ostreatus, and P. cystidiosus, are commercially cultivated in Sri Lanka. Obtaining an efficient, cost-effective, and sustainable substrate is a key factor in promoting the mushroom industry within the country, ensuring export-quality mushroom production. Recently, there have been some attempts to use unconventional substrate materials, such as used diapers, as an innovative and eco-friendly approach, where the high cellulose content of diapers serves as a substrate for fungal growth, while, repurposing problematic non-biodegradable waste streams for agricultural practices. a. [ABSTRACT FROM AUTHOR]

Abstract The present study explored the potential of leaf litter as a source of fungi able to produce ligninolytic enzymes for the biodegradation of anthraquinone dyes. Within the colonies isolated from the leaf litter, only three colonies of two species Trametes were selected based on the detection of oxidation and decolorization halos in Petri dishes with PDA (potato-dextrose-agar) + Guaicol and PDA + RBBR (Remazol Brilliant Blue R). The identification of the colonies was done through sequencing of the ITS region. The enzymatic activity of Lac (lacase), MnP (manganês peroxidase) and LiP (lignina peroxidase) was analyzed by spectrophotometry during fermentation in PD+RBBR imedium. Isolates A1SSI01 and A1SSI02 were identified as Trametes flavida, while A5SS01 was identified as Trametes sp. Laccase showed the highest enzymatic activity, reaching 452.13 IU.L-1 (A1SSI01, 0.05% RBBR) after 96h. Isolate A1SSI02 reached the highest percentage of decolorization, achieving 89.28% in seven days. The results imply that these Trametes isolates can be highly effective in waste treatment systems containing toxic anthraquinone dyes. Keywords: laccase, peroxidases, basidiomycete, litter and biodecolorization.

Background: Ganoderma boninense is a phytopathogen of oil palm, causing basal and upper stem rot diseases. Methods: The genome sequence was used as a reference to study gene expression during growth in a starved carbon (C) and nitrogen (N) environment with minimal sugar and sawdust as initial energy sources. This study was conducted to mimic possible limitations of the C–N nutrient sources during the growth of G. boninense in oil palm plantations. Results: Genome sequencing of an isolate collected from a palm tree in West Malaysia generated an assembly of 67.12 Mb encoding 19,851 predicted genes. Transcriptomic analysis from a time course experiment during growth in this starvation media identified differentially expressed genes (DEGs) that were found to be associated with 29 metabolic pathways. During the active growth phase, 26 DEGs were related to four pathways, including secondary metabolite biosynthesis, carbohydrate metabolism, glycan metabolism and mycotoxin biosynthesis. G. boninense genes involved in the carbohydrate metabolism pathway that contribute to the degradation of plant cell walls were up-regulated. Interestingly, several genes associated with the mycotoxin biosynthesis pathway were identified as playing a possible role in pathogen-host interaction. In addition, metabolomics analysis revealed six metabolites, maltose, xylobiose, glucooligosaccharide, glycylproline, dimethylfumaric acid and arabitol that were up-regulated on Day2 of the time course experiment. Conclusions: This study provides information on genes expressed by G. boninense in metabolic pathways that may play a role in the initial infection of the host. [ABSTRACT FROM AUTHOR]

A comprehensive analysis to survey heme-binding proteins produced by the white-rot fungus Phanerochaete chrysosporium was achieved using a biotinylated heme–streptavidin beads system. Mitochondrial citrate synthase (PcCS), glyceraldehyde 3-phosphate dehydrogenase (PcGAPDH), and 2-Cys thioredoxin peroxidase (mammalian HBP23 homolog) were identified as putative heme-binding proteins. Among these, PcCS and PcGAPDH were further characterized using heterologously expressed recombinant proteins. Difference spectra of PcCS titrated with hemin exhibited an increase in the Soret absorbance at 414 nm, suggesting that the axial ligand of the heme is a His residue. The activity of PcCS was strongly inhibited by hemin with Ki oxaloacetate of 8.7 μM and Ki acetyl-CoA of 5.8 μM. Since the final step of heme biosynthesis occurred at the mitochondrial inner membrane, the inhibition of PcCS by heme is thought to be a physiological event. The inhibitory mode of the heme was similar to that of CoA analogues, suggesting that heme binds to PcCS at His347 at the AcCoA–CoA binding site, which was supported by the homology model of PcCS. PcGAPDH was also inhibited by heme, with a lower concentration than that for PcCS. This might be caused by the different location of these enzymes. From the integration of these phenomena, it was concluded that metabolic regulations by heme in the central metabolic and heme synthetic pathways occurred in the mitochondria and cytosol. This novel pathway crosstalk between the central metabolic and heme biosynthetic pathways, via a heme molecule, is important in regulating the metabolic balance (heme synthesis, ATP synthesis, flux balance of the tricarboxylic acid (TCA) cycle and cellular redox balance (NADPH production) during fungal aromatic degradation. Key points: • A comprehensive survey of heme-binding proteins in P. chrysosporium was achieved. • Several heme-binding proteins including CS and GAPDH were identified. • A novel metabolic regulation by heme in the central metabolic pathways was found. [ABSTRACT FROM AUTHOR]

The Tremellomycetes are a species-rich group within the basidiomycete fungi; however, most analyses of this group to date have focused on pathogenic Cryptococcus species within the order Tremellales. Recent genome-assisted studies of other Tremellomycetes have identified interesting features with respect to biotechnological applications as well as the evolution of genes involved in mating and sexual development. Here, we report genome sequences of two strains of Filobasidium floriforme, a species from the order Filobasidiales, which branches basally to the Tremellales, Trichosporonales, and Holtermanniales. The assembled genomes of strains CBS6241 and CBS6242 are 27.4 Mb and 26.4 Mb in size, respectively, with 8314 and 7695 predicted protein-coding genes. Overall sequence identity at nucleic acid level between the strains is 97%. Among the predicted genes are pheromone precursor and pheromone receptor genes as well as two genes encoding homedomain (HD) transcription factors, which are predicted to be part of the mating type (MAT) locus. Sequence analysis indicates that CBS6241 and CBS6242 carry different alleles for both the pheromone/receptor genes as well as the HD transcription factors. Orthology inference identified 1482 orthogroups exclusively found in F. floriforme, some of which were involved in carbohydrate transport and metabolism. Subsequent CAZyme repertoire characterization identified 267 and 247 enzymes for CBS6241 and CBS6242, respectively, the second highest number of CAZymes among the analyzed Tremellomycete species. In addition, F. floriforme contains five CAZymes absent in other species and several plant-cell-wall degrading CAZymes with the highest copy number in Tremellomycota, indicating the biotechnological potential of this species.

Sesquiterpenes are a type of abundant natural product with widespread applications in several industries. They are biosynthesized by sesquiterpene synthases (STSs). As valuable and abundant biological resources, mushroom-forming fungi are rich in new sesquiterpenes and STSs, which remain largely unexploited. In the present study, we collected information on 172 STSs from mushroom-forming fungi with experimentally characterized products from the literature and sorted them to develop a dataset. Furthermore, we analyzed and discussed the phylogenetic tree, catalytic products, and conserved motifs of STSs. Phylogenetic analysis revealed that the STSs were clustered into four clades. Furthermore, their cyclization reaction mechanism was divided into four corresponding categories. This database was used to predict 12 putative STS genes from the edible fungi Flammulina velutipes. Finally, three FvSTSs were selected to experimentally characterize their functions. FvSTS03 predominantly produced Δ-cadinol and FvSTS08 synthesized β-barbatene as the main product; these findings were consistent with those of the functional prediction analysis. A product titer of 78.8 mg/L β-barbatene was achieved in Saccharomyces cerevisiae via metabolic engineering. Our study findings will help screen or design STSs from fungi with specific product profiles as functional elements for applications in synthetic biology. [ABSTRACT FROM AUTHOR]

Abstract Background The termite-fungus symbiosis is an ancient stable mutualism of two partners that reproduce and disperse independently. With the founding of each termite colony the symbiotic association must be re-established with a new fungus partner. Complementarity in the ability to break down plant substrate may help to stabilize this symbiosis despite horizontal symbiont transmission. An alternative, non-exclusive, hypothesis is that a reduced rate of evolution may contribute to stabilize the symbiosis, the so-called Red King Effect. Methods To explore this concept, we produced the first linkage map of a species of Termitomyces, using genotyping by sequencing (GBS) of 88 homokaryotic offspring. We constructed a highly contiguous genome assembly using PacBio data and a de-novo evidence-based annotation. This improved genome assembly and linkage map allowed for examination of the recombination landscape and its potential effect on the mutualistic lifestyle. Results Our linkage map resulted in a genome-wide recombination rate of 22 cM/Mb, lower than that of other related fungi. However, the total map length of 1370 cM was similar to that of other related fungi. Conclusions The apparently decreased rate of recombination is primarily due to genome expansion of islands of gene-poor repetitive sequences. This study highlights the importance of inclusion of genomic context in cross-species comparisons of recombination rate.

Heterologous production of fungal ligninolytic cocktails is challenging due to the low yields of catalytically active lignin modifying peroxidases. Production using a natural system, such as a wood-rotting fungus, is a promising alternative if specific or preferential induction of the ligninolytic activities could be achieved. Using transcriptomics, gene expression of the white-rot Dichomitus squalens during growth on mixtures of aromatic compounds, with ring structures representing the two major lignin sub-units, was compared to a wood substrate. Most of the genes encoding lignin modifying enzymes (laccases and peroxidases) categorised as highly or moderately expressed on wood were expressed similarly on aromatic compounds. Higher expression levels of a subset of manganese and versatile peroxidases was observed on di- compared to mono-methoxylated aromatics. The expression of polysaccharide degrading enzymes was lower on aromatic compounds compared to wood, demonstrating that the induction of lignin modifying enzymes became more specific. This study suggests potential for aromatic waste streams, e.g. from lignocellulose pretreatment, to produce a lignin-specific enzyme cocktail from D. squalens or other white-rot fungi.

Some of the goals of meat production systems are to improve feed efficiency, reduce costs, provide proper nutrition and avoid metabolic disorders. Hence, the aim of this work was to compare a rabbit diet based on untreated sorghum (T1) with a second one that included the cereal biotransformed by the co-cultivation of two basidiomycetes (T2). Their effects on the cereal, diets, and fecal structural fiber composition, as well as on rabbit intestinal morphology, were evaluated. A completely randomized design was applied, employing 24 New Zealand rabbits (21 days old). The fiber chemical composition results indicated that the sorghum treated with the co-culture had lower amounts of neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, hemicellulose and cellulose, as did rabbits fed the T2 diet. Decreases in the NDF, ADF and cellulose contents were detected in T2-nourished rabbit excrements at day 21 (p ≤ 0.05), while lignin percentages diminished at days 35 and 49 (p ≤ 0.05). These rabbits also showed longer jejunal and cecal villi (p ≤ 0.05). The results obtained suggest a positive effect on the bioavailability of structural fibers after sorghum fungal co-fermentation, as their content decreased in rabbit fecal matter without damaging the intestinal morphology.

Abstract Chemical investigation on the medicinal fungus Ganoderma australe led to the identification of ten new nor-lanostane triterpenes, namely two hexa-nor ones, ganoaustratetraenones A (1) and B (2), five penta-nor ones, ganoaustraldehydes A–E (3–7), and three tetra-nor ones ganoaustrenoic acids A–C (8–10). The chemical structures along with the absolute configurations were determined by extensive spectroscopic analysis of 1D & 2D NMR and HRESIMS data. The postulated biosynthesis pathways of these compounds were proposed. Ganoaustraldehydes A (3) and B (4) showed moderate inhibition against nitric oxide production in RAW264.7 macrophage cells with the respective IC50 values of 32.5, 34.2 µM (the IC50 of positive control pyrrolidine dithiocarbamate was 20.0 µM). Graphical Abstract

Lentinus crinitus mushrooms are consumed by Indigenous Brazilians from the Amazon rainforest and are one of the sources of panepoxydone, a fungal metabolite with several biological activities. However, there are very few reports about L. crinitus mushroom production. This study aimed to evaluate carbon-to-nitrogen (C/N) ratios and nitrogen contents in sugarcane bagasse and soybean meal substrate formulations on mycelial growth, laccase activity, and mushroom production of L. crinitus in solid-state cultivation. Laccase activity of colonized substrate was determined by ABTS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] oxidation method. Mycelial growth, mushroom yield and number, biological efficiency, and biological efficiency-to-cultivation time ratio of L. crinitus were higher when cultivated in the substrate with 1.2% nitrogen (C/N ratio of 36.6). Mushroom mass is the same regardless of the substrate nitrogen content. The substrate colonization lasts 19 days and mushroom harvest 17 days, including primordium formation. The nitrogen ⩾ 1.8% (C/N ratio ⩽ 23.7) in the substrate reduces mycelial growth and completely inhibits mushroom production. However, the increase in nitrogen (⩾ 1.2%) in the substrate increased the laccase activity more than 3.3-times than control (0.9% N); being that laccase activity was negatively correlated with mushroom production and the highest laccase activity was at the fifth cultivation day. This study is the first evidence of the effects of nitrogen content on the substrate in order to produce laccase, mycelial growth, and mushrooms of L. crinitus in solid-state cultivation. The results provide information to support the main conditions to grow and produce L. crinitus mushrooms. [ABSTRACT FROM AUTHOR]

Background: The termite-fungus symbiosis is an ancient stable mutualism of two partners that reproduce and disperse independently. With the founding of each termite colony the symbiotic association must be re-established with a new fungus partner. Complementarity in the ability to break down plant substrate may help to stabilize this symbiosis despite horizontal symbiont transmission. An alternative, non-exclusive, hypothesis is that a reduced rate of evolution may contribute to stabilize the symbiosis, the so-called Red King Effect. Methods: To explore this concept, we produced the first linkage map of a species of Termitomyces, using genotyping by sequencing (GBS) of 88 homokaryotic offspring. We constructed a highly contiguous genome assembly using PacBio data and a de-novo evidence-based annotation. This improved genome assembly and linkage map allowed for examination of the recombination landscape and its potential effect on the mutualistic lifestyle. Results: Our linkage map resulted in a genome-wide recombination rate of 22 cM/Mb, lower than that of other related fungi. However, the total map length of 1370 cM was similar to that of other related fungi. Conclusions: The apparently decreased rate of recombination is primarily due to genome expansion of islands of gene-poor repetitive sequences. This study highlights the importance of inclusion of genomic context in cross-species comparisons of recombination rate. [ABSTRACT FROM AUTHOR]

The mycelial biomass of basidiomycetes is a promising source of compounds and represents an alternative for industrial and biotechnological applications. Fungi use light as information and hold photoresponse mechanisms, in which sensors respond to light wavelengths and regulate various biological processes. Therefore, this study aimed to investigate the effects of blue, green, and red lights on the growth, chemical composition, and antioxidant and antimicrobial activity of Lentinus crinitus mycelial biomass. The chemical composition of the mycelial biomass was determined by chromatographic methods, antioxidant activity was analyzed by in vitro assays, and antimicrobial activity was investigated by the microdilution assay. The highest mycelial biomass yield was observed under blue-light cultivation. Many primordia arose under blue or green light, whereas the stroma was formed under red light. The presence of light altered the primary fungal metabolism, increasing the carbohydrate, tocopherol, fatty acid, and soluble sugar contents, mostly mannitol, and reducing the protein and organic acid concentrations. Cultivation under red light increased the phenol concentration. In contrast, cultivation under blue and green lights decreased phenol concentration. Benzoic and gallic acids were the main phenolic acids in the hydroalcoholic extracts, and the latter acids increased in all cultures under light, especially red light. Mycelial biomass cultivated under red light showed the highest antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The ferric reducing antioxidant power (FRAP) method showed that all light wavelengths increased the antioxidant activity of mycelial biomass, with the highest value under red light. Moreover, the β-carotene/linoleic acid co-oxidation (BCLA) assay demonstrated that the antioxidant activity was affected by light cultivation. Mycelial biomass grown under all conditions exhibited antibacterial and antifungal activities. Thus, mycelial biomass cultivation of L. crinitus under light conditions may be a promising strategy for controlling the mycelial chemical composition and biomass yield. [ABSTRACT FROM AUTHOR]

Lignocellulosic plant biomass is an important feedstock for bio-based economy. In particular, it is an abundant renewable source of aromatic compounds, which are present as part of lignin, as side-groups of xylan and pectin, and in other forms, such as tannins. As filamentous fungi are the main organisms that modify and degrade lignocellulose, they have developed a versatile metabolism to convert the aromatic compounds that are toxic at relatively low concentrations to less toxic ones. During this process, fungi form metabolites some of which represent high-value platform chemicals or important chemical building blocks, such as benzoic, vanillic, and protocatechuic acid. Especially basidiomycete white-rot fungi with unique ability to degrade the recalcitrant lignin polymer are expected to perform highly efficient enzymatic conversions of aromatic compounds, thus having huge potential for biotechnological exploitation. However, the aromatic metabolism of basidiomycete fungi is poorly studied and knowledge on them is based on the combined results of studies in variety of species, leaving the overall picture in each organism unclear. Dichomitus squalens is an efficiently wood-degrading white-rot basidiomycete that produces a diverse set of extracellular enzymes targeted for lignocellulose degradation, including oxidative enzymes that act on lignin. Our recent study showed that several intra- and extracellular aromatic compounds were produced when D. squalens was cultivated on spruce wood, indicating also versatile aromatic metabolic abilities for this species. In order to provide the first molecular level systematic insight into the conversion of plant biomass derived aromatic compounds by basidiomycete fungi, we analyzed the transcriptomes of D. squalens when grown with 10 different lignocellulose-related aromatic monomers. Significant differences for example with respect to the expression of lignocellulose degradation related genes, but also putative genes encoding transporters and catabolic pathway genes were observed between the cultivations supplemented with the different aromatic compounds. The results demonstrate that the transcriptional response of D. squalens is highly dependent on the specific aromatic compounds present suggesting that instead of a common regulatory system, fine-tuned regulation is needed for aromatic metabolism.

Mycelial fungi grow as colonies consisting of polar growing hyphae, developing radially from spore or inoculum. Over time, the colony develops, hyphae are subject to various exogenous or endogenous stimuli, and mycelium becomes heterogeneous in growth, gene expression, biosynthesis, and secretion of proteins and metabolites. Although the biochemical and molecular mechanisms of mycelium heterogeneity have been the subject of many studies, the role of lipids in colony development and zonality is still not understood. This work was undertaken to extend our knowledge of mycelium heterogeneity and to answer the question of how different lipid molecular species are distributed in the surface colony of the basidial fungus Flammulina velutipes and how this distribution correlates with its morphology. The heterogeneity in the lipid metabolism and lipid composition of the fungal mycelium was demonstrated. According to the real-time PCR and LC-MS/MS results, the expression of genes of PC metabolism, accumulation of phospholipid classes, and degree of unsaturation of PC and PE increased in the direction from the center to the periphery of the colony. The peripheral zone of the colony was characterized by a higher value of the PC/PE ratio and a higher level of phospholipids esterified by linolenic acid. Considering that the synthesis of phospholipids in fungi occurs in different ways, we also conducted experiments with deuterium-labeled phospholipid precursors and found out that the Kennedy pathway is the predominant route for PC biosynthesis in F. velutipes. The zonal differences in gene expression and lipid composition can be explained by the participation of membrane lipids in polar growth maintenance and regulation. [ABSTRACT FROM AUTHOR]

The 1-aminocyclopropane-1-carboxylic acid (ACC) pathway that synthesizes ethylene is shared in seed plants, fungi and probably other organisms. However, the evolutionary relationship of the key enzyme ACC oxidase (ACO) in the pathway among organisms remains unknown. Herein, we cloned, expressed and characterized five ACOs from the straw mushroom (Volvariella volvacea) and the oyster mushroom (Pleurotus ostreatus): VvACO1-4 and PoACO. The five mushroom ACOs and the previously identified AbACO of the button mushroom contained all three conserved residues that bound to Fe(II) in plant ACOs. They also had variable residues that were conserved and bound to ascorbate and bicarbonate in plant ACOs and harbored only 1–2 of the five conserved ACO motifs in plant ACOs. Particularly, VvACO2 and AbACO had only one ACO motif 2. Additionally, VvACO4 shared 44.23% sequence identity with the cyanobacterium Hapalosiphon putative functional ACO. Phylogenetic analysis showed that the functional ACOs of monocotyledonous and dicotyledonous plants co-occurred in Type I, Type II and Type III, while putative functional gymnosperm ACOs also appeared in Type III. The putative functional bacterial ACO, functional fungi and slime mold ACOs were clustered in ancestral Type IV. These results indicate that ACO motif 2, ACC and Fe(II) are essential for ACO activity. The ACOs of the other organisms may come from the horizontal transfer of fungal ACOs, which were found ordinarily in basidiomycetes. It is mostly the first case for the horizontal gene transfers from fungi to seed plants. The horizontal transfer of ACOs from fungi to plants probably facilitates the fungal-plant symbioses, plant–land colonization and further evolution to form seeds. [ABSTRACT FROM AUTHOR]

CRISPR/Cas9 has potential for efficient molecular breeding. Recently, a foreign-DNA-free gene-targeting technology was established by introducing a preassembled Cas9 ribonucleoprotein (RNP) complex into the oyster mushroom Pleurotus ostreatus. However, the target gene was restricted to such a gene like pyrG , since screening of a genome-edited strain was indispensable and could be performed via examination of 5-fluoroorotic acid (5-FOA) resistance caused by the disruption of the target gene. In this study, we simultaneously introduced the Cas9 RNP complex targeting fcy1 , a mutation that conferred P. ostreatus resistance to 5-fluorocytosine (5-FC), together with that targeting pyrG. A total of 76 5-FOA resistant strains were isolated during the first screening. Subsequently, a 5-FC resistance examination was conducted, and three strains exhibited resistance. Genomic PCR experiments followed by DNA sequencing revealed that mutations were successfully introduced into fcy1 and pyrG in the three strains. The results indicated that double gene-edited mutants could be obtained in one experiment employing 5-FOA resistance screening for strains with Cas9 RNP incorporation. This work may pave the way for safe CRISPR/Cas9 technology to isolate mutant strains in any gene of interest without an ectopic marker gene. [ABSTRACT FROM AUTHOR]

Celiac disease (CD) is an inflammatory disorder of the small intestine. Gluten peptides are supposed to be responsible for the reaction, the best-researched of which is the so-called '33-mer'. Analogous peptides in secalins (rye) and hordeins (barley) have been described. This study presents the degradation of gliadins, glutenins, hordeins and secalins purified from the respective flours using a prolyl endopeptidase from the Basidiomycete Flammulina velutipes (FvpP). The flour fractions were incubated with the enzyme, and the cleavage sites were determined using high-resolution nLC-qTOF-MS/MS. For the wheat samples, eight cleavage sites in the 33-mer peptide were shown, and all of the six described epitopes were successfully cleaved. For the commercially available prolyl-specific endopeptidase from Aspergillus niger (An-Pep), which was used as a control, only two cleavage sites that cleaved three of the six epitopes were identified. For the secalins, four prolyl-specific cleavage sites in the CD-active peptide QPFPQPQQPIPQ were found for the FvpP but none for the An-Pep. The CD-active peptide QPFPQPEQPFPW in C-hordein was cleaved at three prolyl-specific positions by the FvpP. The study proves the usability of FvpP to degrade CD-inducing peptides in real-grain flour samples and indicates its higher effectiveness compared with An-Pep. A clinical study would be required to assess the therapeutic or preventive potential of FvpP for CD. [ABSTRACT FROM AUTHOR]

Some of the goals of meat production systems are to improve feed efficiency, reduce costs, provide proper nutrition and avoid metabolic disorders. Hence, the aim of this work was to compare a rabbit diet based on untreated sorghum (T1) with a second one that included the cereal biotransformed by the co-cultivation of two basidiomycetes (T2). Their effects on the cereal, diets, and fecal structural fiber composition, as well as on rabbit intestinal morphology, were evaluated. A completely randomized design was applied, employing 24 New Zealand rabbits (21 days old). The fiber chemical composition results indicated that the sorghum treated with the co-culture had lower amounts of neutral detergent fiber (NDF), acid detergent fiber (ADF), lignin, hemicellulose and cellulose, as did rabbits fed the T2 diet. Decreases in the NDF, ADF and cellulose contents were detected in T2-nourished rabbit excrements at day 21 (p ≤ 0.05), while lignin percentages diminished at days 35 and 49 (p ≤ 0.05). These rabbits also showed longer jejunal and cecal villi (p ≤ 0.05). The results obtained suggest a positive effect on the bioavailability of structural fibers after sorghum fungal co-fermentation, as their content decreased in rabbit fecal matter without damaging the intestinal morphology. [ABSTRACT FROM AUTHOR]

Laetiporus sulphureus, a wood-decaying basidiomycete, produces yellow-orange pigments in fruiting bodies and, as was recently shown, in submerged cultivated mycelia. Out of four strains, the most potent laetiporic acid producer was identified and its yield compared in different media. The complex Moser b medium was replaced by potato dextrose broth, achieving higher yields at a lower cost. Cultivation was then scaled up from shake flask to a 7 L stirred tank bioreactor. Optimization of parameters led to increased product concentrations up to 1 g L−1, the highest yield reported so far. An in situ product recovery strategy with a biphasic system was established, increasing the yield by 19% on the shake flask scale. A crude ethanolic extract of the biomass was examined for color stability and application trials. In contrast to what has been suggested in the past, the pigment showed limited long-term stability to oxygen and light, but was stable under storage in the dark at 4 °C under nitrogen. The orange extract was successfully incorporated into different matrices like foods, cosmetics and textiles. Laetiporic acid can potentially replace petrochemical based synthetic dyes, and can thus support the development of a circular bioeconomy. [ABSTRACT FROM AUTHOR]

Chemical investigation on the medicinal fungus Ganoderma australe led to the identification of ten new nor-lanostane triterpenes, namely two hexa-nor ones, ganoaustratetraenones A (1) and B (2), five penta-nor ones, ganoaustraldehydes A–E (3–7), and three tetra-nor ones ganoaustrenoic acids A–C (8–10). The chemical structures along with the absolute configurations were determined by extensive spectroscopic analysis of 1D & 2D NMR and HRESIMS data. The postulated biosynthesis pathways of these compounds were proposed. Ganoaustraldehydes A (3) and B (4) showed moderate inhibition against nitric oxide production in RAW264.7 macrophage cells with the respective IC50 values of 32.5, 34.2 µM (the IC50 of positive control pyrrolidine dithiocarbamate was 20.0 µM). [ABSTRACT FROM AUTHOR]

Genetic variability can be generated by different mechanisms, and across the life cycle. Many basidiomycete fungi have an extended somatic stage, during which each cell carries two genetically distinct haploid nuclei (dikaryosis), resulting from fusion of two compatible monokaryotic individuals. Recent findings have revealed remarkable genome stability at the nucleotide level during dikaryotic growth in these organisms, but whether this pattern extends to mutations affecting large genomic regions remains unknown. Furthermore, despite high genome integrity during dikaryosis, basidiomycete populations are not devoid of genetic diversity, begging the question of when this diversity is introduced. Here, we used a Marasmius oreades fairy ring to investigate the rise of large-scale variants during mono- and dikaryosis. By separating the two nuclear genotypes from four fruiting bodies and generating complete genome assemblies, we gained access to investigate genomic changes of any size. We found that during dikaryotic growth in nature the genome stayed intact, but after separating the nucleotypes into monokaryons, a considerable amount of structural variation started to accumulate, driven to large extent by transposons. Transposon insertions were also found in monokaryotic single-meiospore isolates. Hence, we show that genome integrity in basidiomycetes can be interrupted during monokaryosis, leading to genomic rearrangements and increased activity of transposable elements. We suggest that genetic diversification is disproportionate between life cycle stages in mushroom-forming fungi, so that the short-lived monokaryotic growth stage is more prone to genetic changes than the dikaryotic stage. [ABSTRACT FROM AUTHOR]

Micropsalliota is a relatively small genus containing only 62 previously identified species. Here, we describe six new taxa of Micropsalliota based on morphological and phylogenetic analyses: M. minor, M. ovalispora, M. pseudodelicatula, M. rufosquarrosa, M. tenuipes, and M. wuyishanensis and a newrecord taxon to China. The firstMaximumlikelihood and Bayesian analyses of a three-gene dataset (ITS, LSU, and rpb2) separated the genus into 18 weakly to strongly supported major clades and subclades, but only a few subclades were synapomorphies. According to phylogenetic analyses, M. cornuta does not belong in Micropsalliota. A key to 20 species of Micropsalliota in China is provided. [ABSTRACT FROM AUTHOR]

Sesquiterpenes are a type of abundant natural product with widespread applications in several industries. They are biosynthesized by sesquiterpene synthases (STSs). As valuable and abundant biological resources, mushroom-forming fungi are rich in new sesquiterpenes and STSs, which remain largely unexploited. In the present study, we collected information on 172 STSs from mushroom-forming fungi with experimentally characterized products from the literature and sorted them to develop a dataset. Furthermore, we analyzed and discussed the phylogenetic tree, catalytic products, and conserved motifs of STSs. Phylogenetic analysis revealed that the STSs were clustered into four clades. Furthermore, their cyclization reaction mechanism was divided into four corresponding categories. This database was used to predict 12 putative STS genes from the edible fungi Flammulina velutipes. Finally, three FvSTSs were selected to experimentally characterize their functions. FvSTS03 predominantly produced Δ-cadinol and FvSTS08 synthesized β-barbatene as the main product; these findings were consistent with those of the functional prediction analysis. A product titer of 78.8 mg/L β-barbatene was achieved in Saccharomyces cerevisiae via metabolic engineering. Our study findings will help screen or design STSs from fungi with specific product profiles as functional elements for applications in synthetic biology.

BackgroundConiophora olivacea is a basidiomycete fungus belonging to the order Boletales that produces brown-rot decay on dead wood of conifers. The Boletales order comprises a diverse group of species including saprotrophs and ectomycorrhizal fungi that show important differences in genome size.ResultsIn this study we report the 39.07-megabase (Mb) draft genome assembly and annotation of C. olivacea. A total of 14,928 genes were annotated, including 470 putatively secreted proteins enriched in functions involved in lignocellulose degradation. Using similarity clustering and protein structure prediction we identified a new family of 10 putative lytic polysaccharide monooxygenase genes. This family is conserved in basidiomycota and lacks of previous functional annotation. Further analyses showed that C. olivacea has a low repetitive genome, with 2.91% of repeats and a restrained content of transposable elements (TEs). The annotation of TEs in four related Boletales yielded important differences in repeat content, ranging from 3.94 to 41.17% of the genome size. The distribution of insertion ages of LTR-retrotransposons showed that differential expansions of these repetitive elements have shaped the genome architecture of Boletales over the last 60 million years.ConclusionsConiophora olivacea has a small, compact genome that shows macrosynteny with Coniophora puteana. The functional annotation revealed the enzymatic signature of a canonical brown-rot. The annotation and comparative genomics of transposable elements uncovered their particular contraction in the Coniophora genera, highlighting their role in the differential genome expansions found in Boletales species.

Telomerase is a eukaryotic ribonucleoprotein (RNP) enzyme that adds DNA repeats onto chromosome ends to maintain genomic stability and confer cellular immortality in cancer and stem cells. The telomerase RNA (TER) component is essential for telomerase catalytic activity and provides the template for telomeric DNA synthesis. The biogenesis of TERs is extremely divergent across eukaryotic kingdoms, employing distinct types of transcription machinery and processing pathways. In ciliates and plants, TERs are transcribed by RNA polymerase III (Pol III), while animal and ascomycete fungal TERs are transcribed by RNA Pol II and share biogenesis pathways with small nucleolar RNA (snoRNA) and small nuclear RNA (snRNA), respectively. Here, we report an unprecedented messenger RNA (mRNA)-derived biogenesis pathway for the 1,291 nucleotide TER from the basidiomycete fungus Ustilago maydis. The U. maydis TER (UmTER) contains a 50-monophosphate, distinct from the 50 2,2,7-trimethylguanosine (TMG) cap common to animal and ascomycete fungal TERs. The mature UmTER is processed from the 30-untranslated region (30-UTR) of a larger RNA precursor that possesses characteristics of mRNA including a 50 7-methyl-guanosine (m7G) cap, alternative splicing of introns, and a poly(A) tail. Moreover, this mRNA transcript encodes a protein called Early meiotic induction protein 1 (Emi1) that is conserved across dikaryotic fungi. A recombinant UmTER precursor expressed from an mRNA promoter is processed correctly to yield mature UmTER, confirming an mRNA-processing pathway for producing TER. Our findings expand the plethora of TER biogenesis mechanisms and demonstrate a pathway for producing a functional long noncoding RNA from a protein-coding mRNA precursor. [ABSTRACT FROM AUTHOR]

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

Basidiomycetes can produce various valuable secondary metabolites with bioactivities. However, the low titer of metabolites has greatly limited their application. Currently, shortage of mutant complementation techniques, especially in-situ complementation, impeded the elucidation of their metabolites biosynthesis pathways. In this work, Ganoderma lucidum , a traditional Chinese medicinal basidiomycete, which can produce various bioactive ganoderic acids (triterpenoids), was used; and a clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) mediated in-situ complementation system for two functional genes was established. For a disruptant of gl26097 , which encodes a Cys2-His2 (C2H2)-type zinc finger transcription factor involved in calmodulin-calcineurin signaling, one in-situ complemented strain of 8 transformants was obtained,concluded by sequencing confirmation. For the function of gl26097 , both the cell growth and content of four identified individual ganoderic acids (GAs) were restored in the complemented strain compared to the gl26097 disruptant. For a disruptant of cyp5150l8 , which is responsible for transforming lanosterol to a GA, one in-situ complemented strain from 8 transformants was obtained as confirmed by sequencing. The titer of four identified GAs increased significantly in the complemented strain compared to the cyp5150l8 disruptant. To the best of our knowledge, this is the first research report on applying CRISPR-Cas9 to complement mutants in-situ in basidiomycetes. The CRISPR-Cas9 assisted gene complementation technology will be helpful to study gene functions and GA biosynthesis pathway in G. lucidum , which may be also applied to other basidiomycetes. [Display omitted] • CRISPR-assisted gene in-situ complementation was established in basidiomycete. • Two gene disruptants of G. lucidum were complemented via the developed technology. • The work will be helpful to gene function and biosynthetic pathway study in G. lucidum. [ABSTRACT FROM AUTHOR]

Pucciniales (rust fungi) are one of the largest fungal order of plant pathogens. They collectively infect key crops such as wheat and soybean, and threaten global food security. In the early 2010s, the genome sequences of three rust fungi were released: Melampsora larici-populina (the poplar leaf rust fungus), Puccinia graminis f. sp. tritici (the wheat stem rust fungus), and Puccinia striiformis f. sp. triciti (the wheat stripe rust or wheat yellow rust fungus). The availability of those genomes has forwarded rust biology into the post-genomic era, sparking a series of genomics, transcriptomics, in silico, and functional studies. Here, we snapshot the last 10 years of postgenomics studies addressing M. larici-populina, P. graminis f. sp. tritici, and/or P. striiformis f. sp. tritici. This mini-review notably reveals the model speciescentered structure of the research community, and highlights the drastic increase of the number of functional studies focused on effectors since 2014, which notably revealed chloroplasts as a central host compartment targeted by rust fungi. This mini-review also discusses genomics-facilitated studies in other rust species, and emerging post-genomic research trends related to fully-phased rust genomes. [ABSTRACT FROM AUTHOR]

A rare basidiomycete Quambalaria cyanescens, documented so far on various substrates worldwide, was isolated from the skin of edible frog (Pelophylax esculentus) captured in South Banat. The fungal identification was based on sequencing of ITS region and BLAST analyses. The presence of Q. cyanescens in the amphibian skin microbiome is not only the first finding of this fungus in Serbia but also the recording of new ecological habitat for this rare species of micromycetes. Phylogenetic analyses revealed the high similarity of isolate in this study with foliar pathogens of Eucalyptus in Australia. [ABSTRACT FROM AUTHOR]

Ceriporiopsis subvermispora is a white-rot fungus with great potential for industrial and biotechnological applications, such as the pretreatment of lignocellulose in biorefineries, as it decomposes the lignin in the plant cell wall without causing severe cellulose degradation. A genetic transformation system was recently developed; however, gene-targeting experiments to disrupt or modify the gene(s) of interest remain challenging, and this is a bottleneck for further molecular genetic studies and breeding of C. subvermispora. Herein, we report efficient clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9)-assisted gene mutagenesis in this fungus. Two plasmids expressing Cas9 together with a different pyrG-targeting single-guide RNA were separately introduced into the monokaryotic C. subvermispora strain FP-90031-Sp/1, which frequently generated strains that exhibited resistance to 5-fluoroorotic acid and uridine/uracil auxotrophy. Southern blot analyses and genomic polymerase chain reaction followed by DNA sequencing of some mutants revealed that they were pyrG mutants. We also observed that hygromycin resistance of the pyrG mutants was frequently lost after repeated subcultivations, indicating that a maker-free genome editing occurred successfully. It is also suggested that a gene mutation(s) can be introduced via a transient expression of Cas9 and a single-guide RNA; this feature, together with high-frequency gene targeting using the CRISPR/Cas9 system, would be helpful for studies on lignocellulose-degrading systems in C. subvermispora. Key points: • Efficient plasmid-based CRISPR/Cas9 was established in C. subvermispora. • The mutations can be introduced via a transient expression of Cas9 and sgRNA. • A maker-free CRISPR/Cas9 is established in this fungus. [ABSTRACT FROM AUTHOR]

Due to their enzymatic and bioaccumulation faculties the use of macromycetes for the decontamination of polluted matrices seems reasonable for bioremediation. For this reason, the aim of our study was to evaluate the mycoremediation ability of Agaricus bisporus cultivated on compost mixed with flotation tailings in different quantities (1, 5, 10, 15, and 20% addition). The biomass of the fruit bodies and the content of 51 major and trace elements were determined. Cultivation of A. bisporus in compost moderately polluted with flotation tailings yielded significantly lower (the first flush) and higher (the second flush) biomass of fruit bodies, compared with the control treatment. The presence of toxic trace elements did not cause any visible adverse symptoms for A. bisporus. Increasing the addition of flotation tailings to the compost induced an elevated level of most determined elements. A significant increase in rare earth elements (both flushes) and platinum group elements (first flush only) was observed. The opposite situation was recorded for major essential elements, except for Na and Mg in A. bisporus from the second flush under the most enriched compost (20%). Nevertheless, calculated bioaccumulation factor values showed a selective accumulation capacity—limited for toxic elements (except for Ag, As, and Cd) and the effective accumulation of B, Cu, K, and Se. The obtained results confirmed that A. bisporus can be used for practical application in mycoremediation in the industry although this must be preceded by larger-scale tests. This application seems to be the most favorable for media contaminated with selected elements, whose absorption by fruiting bodies is the most efficient. [ABSTRACT FROM AUTHOR]

In the present study, we investigated whether a non-invasive metabolic heat flux analysis could serve the determination of the functional traits in free-living saprotrophic decomposer fungi and aid the prediction of fungal influences on ecosystem processes. For this, seven fungi, including ascomycete, basidiomycete, and zygomycete species, were investigated in a standardised laboratory environment, employing wheat straw as a globally relevant lignocellulosic substrate. Our study demonstrates that biocalorimetry can be employed successfully to determine growth-related fungal activity parameters, such as apparent maximum growth rates (AMGR), cultivation times until the observable onset of fungal growth at AMGR (tAMGR), quotients formed from the AMGR and tAMGR (herein referred to as competitive growth potential, CGP), and heat yield coefficients (YQ/X), the latter indicating the degree of resource investment into fungal biomass versus other functional attributes. These parameters seem suitable to link fungal potentials for biomass production to corresponding ecological strategies employed during resource utilisation, and therefore may be considered as fungal life history traits. A close connection exists between the CGP and YQ/X values, which suggests an interpretation that relates to fungal life history strategies. [ABSTRACT FROM AUTHOR]

There is an urgent need for improvements in animal production, particularly for ruminants, such that more sustainable and efficient processes are developed for obtaining more nutritious and efficient feeds. Filamentous fungi can add value to residual plant biomass, and may also have the potential to produce metabolites and enrich plant biomasses used in animal nutrition, converting them into nutraceutical sources. Thus, in this work, filamentous fungal fermentation of ruminant feed biomasses commonly used in Brazil was performed, and the enrichment for bioactive metabolites was tested. For this, Fistulina hepatica, Ganoderma lucidum, Pleurotus pulmonarius, Panus lecomtei, and Aspergillus terreus were grown for 28 days on different substrates: starchy grains- (sorghum, oat, and corn), fibrous substrates (coast-cross, rice husk, and moringa plant) and protein-rich substrates (cottonseed cake and pigeon pea plant). Fermented substrates were evaluated for laccase activity, crude protein, β-glucan, and lovastatin content. The highest growth rate was observed for G. lucidum in oat substrate (OT-01) (0.708 ± 0.035 cm/day) and F. hepatica in oat + coast-cross + pigeon pea treatment (OT-10) (0.607 ± 0.012 cm/day). High laccase activity was observed for P. lecomtei grown in starchy grain + moringa + pigeon pea substrate, reaching an activity of 416.8 ± 20.28 U/g. A. terreus growth in ST-09 (sorghum + pigeon pea) showed higher protein (15.3 ± 0.46%), β-glucan (503.56 ± 8.6 mg/g) and lovastatin (1.10 ± 0.17 mg/g) content compared to untreated substrates. These results demonstrate that filamentous fungi are an alternative for nutraceutical enrichment of ruminant feed biomasses. To the best of our knowledge, this is the first report in which P. lecomtei and F. hepatica are evaluated for their ability to be cultivated in ruminant feed substrates from Brazil. [ABSTRACT FROM AUTHOR]

We report an annotated draft genome of Heterobasidion occidentale , a fungus (Basidiomycota, Agaricomycetes) that has pathogenic and saprophytic lifestyles. This fungus belongs to the H. annosum (Fr.) Bref. sensu lato species complex that comprises several root rot pathogens. Heterobasidion occidentale causes annosus root and butt rot primarily in true fir ( Abies spp.) and spruce ( Picea spp.) species throughout western North America., Competing Interests: The authors declare no conflict of interest.

Wood-decaying white rot fungi live in changing environmental conditions and may switch from aerobic to fermentative metabolism under oxygen depletion. Decomposition of wood and lignocellulose by fungi is dependent on enzymatic and oxidative biochemistry including generation of reactive oxygen species. In this study, we subjected semi-solid wood-substrate cultures of the white rot fungus Phlebia radiata to oxidative stress by addition of hydrogen peroxide under aerobic and anaerobic cultivation conditions. Wood decomposition and fungal metabolism were followed by analysis of extracellular organic compounds, mycelial growth, and laccase activity. Under both atmospheric conditions, accumulation of bioactive aromatic compounds from birch wood occurred into the culture supernatants after hydrogen peroxide treatment. The supernatants inhibited both fungal growth and laccase activity. However, the fungus recovered from the oxidative stress quickly in a few days, especially when cultivated under regular aerobic conditions. With repeated hydrogen peroxide treatments, laccase suppressive-recovery effect was observed. Culture supernatants demonstrated antioxidant and antimicrobial effects, in concert with emergence of chlorinated birch-derived organic compounds. Bioactivities in the cultures disappeared in the same pace as the chlorinated compounds were transformed and de-chlorinated by the fungus. Our results indicate tolerance of white rot fungi against excessive oxidative stress and wood-derived, growth-inhibiting and harmful agents., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Peritoneal dialysis (PD)-associated fungal peritonitis, although rare, presents significant challenges in diagnosis and management. Here, we present the first case of PD-related peritonitis attributed to Psathyrella candolleana and highlight a potential route of infection through contamination from the PD catheter belt. A 37-year-old female, with a history of heart and lung transplantation and undergoing continuous ambulatory PD, presented with acute abdominal pain and cloudy PD effluent (PDE). Genetic analysis of PDE and PD catheter tip confirmed diagnosis of P. candolleana . Treatment was successful without any relapses with timely PD catheter removal and an extended course of antifungal therapy. The root cause analysis suspected the dirt-stained PD catheter belt as the origin of contamination. In conclusion, this is the first case of P. candolleana infection in PD-related peritonitis. Preventive strategies should prioritize hygiene practices, including the PD belt to mitigate the risk of contamination and subsequent infections of such pathogens.

Pucciniales (rust fungi) are one of the largest fungal order of plant pathogens. They collectively infect key crops such as wheat and soybean, and threaten global food security. In the early 2010s, the genome sequences of three rust fungi were released: Melampsora larici-populina (the poplar leaf rust fungus), Puccinia graminis f. sp. tritici (the wheat stem rust fungus), and Puccinia striiformis f. sp. triciti (the wheat stripe rust or wheat yellow rust fungus). The availability of those genomes has forwarded rust biology into the post-genomic era, sparking a series of genomics, transcriptomics, in silico, and functional studies. Here, we snapshot the last 10 years of post-genomics studies addressing M. larici-populina, P. graminis f. sp. tritici, and/or P. striiformis f. sp. tritici. This mini-review notably reveals the model species-centered structure of the research community, and highlights the drastic increase of the number of functional studies focused on effectors since 2014, which notably revealed chloroplasts as a central host compartment targeted by rust fungi. This mini-review also discusses genomics-facilitated studies in other rust species, and emerging post-genomic research trends related to fully-phased rust genomes.

Environmental pollutants are today a major concern and an intensely discussed topic on the global agenda for sustainable development. They include a wide range of organic compounds, such as pharmaceutical waste, pesticides, plastics, and volatile organic compounds that can be found in air, soil, water bodies, sewage, and industrial wastewater. In addition to impacting fauna, flora, and fungi, skin absorption, inhalation, and ingestion of some pollutants can also negatively affect human health. Fungi play a crucial role in the decomposition and cycle of natural and synthetic substances. They exhibit a variety of growth, metabolic, morphological, and reproductive strategies and can be found in association with animals, plants, algae, and cyanobacteria. There are fungal strains that occur naturally in soil, sediment, and water that have inherent abilities to survive with contaminants, making the organism important for bioassay applications. In this context, we reviewed the applications of fungal-based bioassays as a versatile tool for environmental monitoring.

Lepista sordida is a fairy ring-forming fungus that belongs to the family Tricholomataceae and is widely distributed in the Northern Hemisphere. Here, we report the complete mitochondrial genome sequence of L. sordida. The mitochondrial genome (57,375 bp) contained 20 protein-coding genes, 2 ribosomal RNA genes, and 26 transfer RNA genes. Phylogenetic analysis based on 14 conserved protein sequences from L. sordida and 15 related basidiomycetes showed that L. sordida was located on the outermost branch of the Tricholomataceae clade. This study is the first to report the complete mitochondrial genome sequence of a fairy ring-forming fungus belonging to the genus Lepista.

The Amazon rainforest has a rich biodiversity, and studies of Basidiomycete fungi that have biomolecules of biotechnological interest are relevant. The use of lignocellulosic biomass in biotechnological processes proposes an alternative use, and also adds value to the material when employed in the bioconversion of agro-industrial waste. In this context, this study evaluate the production of lignocellulolytic enzymes (carboxymethylcellulases (CMCase), xylanase, pectinase, laccase) as well as phenolic compounds and proteases by solid-state fermentation (SSF) using the fungus Lentinus strigosus isolated from Amazon. The guarana (Paullinia cupana) residue was characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). SSF was carried out with 60% humidification of the residue, at 30 °C, for 10 days. The lignocellulosic biomass presented fragmented structures with irregular shapes and porosities, and was mainly constituted by cellulose (19.16%), hemicellulose (32.83%), and lignin (6.06%). During the SSF, significant values of CMCase (0.84 U/g) on the 8th day, xylanase (1.00 U/g) on the 7th day, pectinase (2.19 U/g) on the 6th day, laccase (176.23 U/mL) on the 5th day, phenolic compounds (10.27 μg/mL) on the 1st day, soluble proteins (0.08 mg/mL) on the 5th day, and protease (8.30 U/mL) on the 6th day were observed. In general, the agro-industrial residue used provided promising results as a viable alternative for use as a substrate in biotechnological processes. [ABSTRACT FROM AUTHOR]

Iugisporipsathyra, a new psathyrelloid genus from tropical red soil of China, is established with I. reticulopilea as the type species. The new genus is characterised by basidiomata psathyrelloid, pileus rugose to appearing reticulate ridged, covered by persistent, but inconspicuous villus, pleurocystidia absent and ridge-ornamented spores with an obvious suprahilar plage. The genus is unique amongst Psathyrellaceae in producing ridge-ornamented spores with an obvious suprahilar plage and forms a distinct lineage within Psathyrellaceae, based on the Maximum Likelihood and Bayesian Inference analyses of a combined three-gene sequence dataset (ITS, LSU and β-tub). Full descriptions and photographs of the new genus and species are presented. [ABSTRACT FROM AUTHOR]

• Two genes essential for basidiospore production were identified in P. ostreatus. • Mutations in pcl1 or cro6c impair post-meiotic processes. • They would be promising targets for breeding sporeless mushrooms. A sporeless strain is an important breeding target in the mushroom industry. However, basidiospore production in the oyster mushroom Pleurotus ostreatus has been shown to be impaired by single-gene mutations in only two meiosis-related genes, mer3 and msh4. This study proposed a strategy for identifying the genes essential for basidiospore formation after meiotic division to determine new targets for molecular breeding. RNA-seq analysis was performed to identify P. ostreatus genes that are specifically expressed in the gill tissue of fruiting bodies, where basidiospore formation occurs. Transcriptome data during fruiting development of Coprinopsis cinerea , in which the meiotic steps progress synchronously, were then used to identify genes that are active in the postmeiotic stages. Based on these comparative analyses, five P. ostreatus genes were identified. Plasmids containing expression cassettes for hygromycin B-resistance screening, Cas9, and single-guide RNA targeting each gene were introduced into the protoplasts of dikaryotic strain, PC9×#64, to generate dikaryotic gene disruptants. Among the obtained transformants, three dikaryotic pcl1 disruptants and two cro6c disruptants did not produce basidiospores. Microscopic analyses indicated that spore formation was arrested at particular stages in these gene disruptants. These results indicate that these two genes are essential for mature spore formation in this fungus. [ABSTRACT FROM AUTHOR]