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Agaricus bisporus is in general cultivated on wheat and rice straw in China. However, millet straw is a potential alternative resource for Agaricus bisporus cultivation, but this has hardly been studied. In the present study, the feasibility of millet straw based mushroom production was analyzed by three successive trials. Mature compost demonstrated high quality with total nitrogen, pH, and C/N ratio of 2.0%, 7.5, and 18:1 respectively, which was suitable for mushroom mycelia growth. During composting, 47-50% of cellulose, 63-65% of hemicellulose, and 8-17% lignin were degraded, while 22-27% of cellulose, 14-16% of hemicellulose, and 15-21% of lignin were consumed by A. bisporus mycelia during cultivation. The highest FPUase and CMCase were observed during mushroom flushes. Endo-xylanase had the key role in hemicellulose degradation with high enzyme activity during cultivation stages. Laccase participated in lignin degradation with the highest enzyme activity in Pinning stage followed by a sharp decline at the first flush. Yield was up to 20 kg/m 2 , as this is similar to growth on wheat straw, this shows that millet straw is an effective resource for mushroom cultivation. Actinobacteria, Bacteroidetes, Chloroflexi, Deinococcus-Thermus, Firmicutes, and Proteobacteria were the dominant phyla, based on 16S rRNA gene sequencing during composting. The key environmental factors dominating bacterial communities of the samples were determined to be pH value, cellulose content, and hemicellulose content for prewetting and premixed phase of basic mixture (P0); moisture content for phase I (PI); and nitrogen content, lignin content, and ash content for phase II (PII), respectively.

Agaricus brasiliensis is popular because of its nutritional and medicinal properties. Brazil supplies this edible mushroom to international markets, where quality standards are very restrictive regarding the allowable concentrations of heavy metals, especially cadmium. In order to evaluate the accumulation of cadmium, chromium, and lead in A. brasiliensis, the fungus was grown as producers cultivate it, using plastic bags containing inoculated compost with soil as a covering. We analyzed the soil and compost using atomic absorption spectrophotometry to determine their physical and chemical properties and natural concentrations of the heavy metals cadmium, chromium, and lead. In addition to the metals naturally found in the soil and compost, 6.6 mgkg-1 cadmium chloride, 13.2 mgkg-1 chromium sulfate, and 1.2 mgkg-1 lead nitrate were added to the soil covering to simulate contaminated soils and to allow us to evaluate potential adsorption by the fungus. An analysis of the fruiting bodies after fructification showed accumulation of the heavy metals, with the greatest amount in those fruiting bodies that had been contaminated with cadmium. Furthermore, the contaminated fruiting bodies had the least raw protein content, were physically deformed, and exhibited variations in the results of Fourier transform infrared spectroscopy analyses.

We show that a lectin like protein from the mushroom Agaricus bisporus (LSMT) is capable to permeate the epithelial monolayer barrier of the intestine ex vivo. The protein is not toxic or immunogenic upon prolonged administration and elevated dose in mice. Thus, it could be a candidate as a drug carrier for oral administration. However, its permeability should be tested after the protein has been modified, mimicking the condition in which it is used as a drug carrier. The protein was conjugated to captopril, the selected model of a Biopharmaceutical Classification System (BCS) class III drug, with high solubility but poor permeability. The drug was conjugated to LSMT that had been modified with 4-succinimidyloxycarbonyl-alpha-methyl-2-pyridyldithiotoluene (SMPT) as a linker. The success of LSMT modification was confirmed with TLC and MS; the latter also indicated the amount of captopril molecule linked. The modified LSMT could permeate through the intestinal monolayer barrier, and thus could be absorbed in the intestine after modification. The modified protein appears to remain stable after incubation in simulated gastrointestinal fluids. This pioneering work provides an essential basis for further development of the protein as a drug carrier for oral administration.

Intensive rice-wheat cultivation cycle in Northern belt of India in general and in the State of Punjab in particular results in large volumes of straw and other post-harvest residue annually. The agricultural area, bordering the districts of Nawanshahr and Hoshiarpur, is popularly known as the seleniferous belt of India. The agri-residues, generated in seleniferous region of this state, are observed to contain significantly high concentration of selenium (Se). The present study was aimed to evaluate the Se uptake by different mushroom species: Pleurotus sajor-caju, Pleurotus ostreatus, Pleurotus citrinopileatus, Agaricus bisporus, and Volvariella volvacea, cultivated on Se-rich wheat and paddy straw from the seleniferous region. Wheat (Pleurotus species and A. bisporus) and paddy straw (V. volvacea) was inoculated with the mycelium spawn and left for 7-20 days, depending on the species, to grow. Control mushrooms were grown analogously using the agricultural residues from non-seleniferous area of the State of Punjab. All fruiting bodies were collected and analyzed in triplicate. Se was quantified using inductively coupled plasma sector field mass spectrometry. The Se accumulation was high in all species under study, being the highest in A. bisporus (1396 μg/g vs. 46.8 μg/g in controls - dry weight) and V. volvacea (231 μg/g vs. 3.77 μg/g - dry weight). The observed biological efficiency and total yield for all mushroom species showed good and unaltered productivity in Se-rich conditions, if compared to the controls. The Se-rich mushrooms can be prospective Se-supplements sourcing and biofortified foods, providing readily bioavailable and accessible Se for the diets deficient of this biologically essential element., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Wild and cultured mushrooms have been extensively used for food and medicinal purposes all around the world. However, there is limited information on chemical composition, health enhancing effects and contributions on diet of some mushrooms (e.g., Agaricus arvensis) widely distributed in many countries including United Kingdom, Australia, Turkey etc. Therefore, the present study was aimed to analyse the bioactive composition and ameliorative effects of A. arvensis via evaluating in vitro and in vivo antioxidant properties in CCl 4 induced rat model. The extract exhibited higher antioxidant capacities in vitro than that of the positive control (Reishi-Shiitake-Maitake standardized extract). Administration of the extract had significant regulative effects in the levels of AST, ALT, LDH, Urea and TRIG levels according to CCl 4 group. Additionally, lipid peroxidation and GSH in the brain, kidney and liver tissues was regulated by extract treated groups compared to the CCI 4 group. The supplementation of the extract at the dose of 100 mg/kg regulated the levels of GST, GR, CAT and GPx enzyme activities in brain and liver, but not in kidney tissue. There was approximately three fold increase in CAT enzyme activity in kidney tissue of extract treated groups compared to Control and CCl 4 groups. The extract contained a rich composition of bioactive compounds including phenolics (protocatechuic acid and p-hydroxybenzoic acid), volatile compounds (benzaldehyde, palmitic acid and linoleic acid) and mineral compounds (K, Si, Mg and Na). Data obtained within this study suggests that A. arvensis might be used for food industries in order to obtain nutritional products.

Three culinary-medicinal fungi and mushrooms (Agaricus bisporus AS2796, Helvella lacunosa X1 and Fomitiporia yanbeiensis S. Guo & L. Zhou) were individually inoculated into different cereal grains (wheat, rice, oat, corn, millet, quinoa, buckwheat, soybean, pea and sorghum) and the antioxidant properties of fungus-fermented products after solid-state fermentation (SSF) (0, 7, 14, 21, 28 and 35 days; 25°C) were studied. The results showed that the total phenol contents (TPCs) of the fermented cereals varied with fermentation time and the starter organisms. According to the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power, ferrous ion chelating ability and superoxide anion radical scavenging ability of ethanolic extracts from the fungus-fermented products (35 days), it was shown that the antioxidant properties of all the products were significantly stronger than uninoculated grains. It revealed that SSF on cereal grains by dietary fungi is a biotechnological strategy, which may enhance the antioxidant properties of the substrate. The three medicinal mushroom and fungi-fermented products were relatively effective in the antioxidant properties assayed and might be potential antioxidants for application in food products.

We examined the effect of various concentrations of HAuCl 4 , AgNO 3 , Na 2 SeO 3 , Na 2 SiO 3 , and GeO 2 on mycelial growth of the soil basidiomycetes Agaricus bisporus and A. arvensis in submerged and solid media. Fungal mycelial extracts and cell-free culture filtrates were able to reduce ions of Au, Ag, Se, Si, and Ge compounds, forming Au 0 , Ag 0 , Se 0 , Si 0 /SiO 2 and Ge 0 /GeO 2 nanoparticles. The physical characteristics of the mycogenic nanoparticles differed depending on the species of Agaricus and the type of extract. Au nanospheres obtained with cell-free culture filtrates were of 2-5 nm diameter in A. bisporus and of 2-10 nm in A. arvensis. Nanoparticles produced by extracts of mycelia were several times larger and highly heterogenous. Ag nanoparticles produced by cell-free culture filtrates were spherical or irregular-shaped and agglomerated, whereas with extracts of mycelia, small homogenous nanospheres of 1-10 nm were formed. Se nanospheres obtained with cell-free culture filtrates were of 100-250 nm diameter in A. bisporus and of 150-550 nm diameter in A. arvensis. The particles synthesized with extracts of mycelia were of 40-140 nm in A. bisporus and of 100-250 nm in A. arvensis. Incubation of Na 2 SiO 3 with cell-free culture filtrates resulted in porous Si nanoparticles of 30-65 nm in A. bisporus and of 50-200 nm in A. arvensis. Ge nanoparticles synthesized by both Agaricus species were mostly spheres of 50-250 nm diameter., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The degradation of two isomeric three-ringed polycyclic aromatic hydrocarbons by the white rot fungus Pleurotus ostreatus D1 and the litter-decomposing fungus Agaricus bisporus F-8 was studied. Despite some differences, the degradation of phenanthrene and anthracene followed the same scheme, forming quinone metabolites at the first stage. The further fate of these metabolites was determined by the composition of the ligninolytic enzyme complexes of the fungi. The quinone metabolites of phenanthrene and anthracene produced in the presence of only laccase were observed to accumulate, whereas those formed in presence of laccase and versatile peroxidase were metabolized further to form products that were further included in basal metabolism (e.g. phthalic acid). Laccase can catalyze the initial attack on the PAH molecule, which leads to the formation of quinones, and that peroxidase ensures their further oxidation, which eventually leads to PAH mineralization. A. bisporus, which produced only laccase, metabolized phenanthrene and anthracene to give the corresponding quinones as the dominant metabolites. No products of further utilization of these compounds were detected. Thus, the fungi's affiliation with different ecophysiological groups and their cultivation conditions affect the composition and dynamics of production of the ligninolytic enzyme complex and the completeness of PAH utilization., (Copyright © 2018 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

We demonstrate the feasibility of a self-propelling mushroom motor, namely a 'logibot', as a functional unit for the construction of a host of optimized binary logic gates. Emulating the chemokinesis of unicellular prokaryotes or eukaryotes, the logibots made stimuli responsive conditional movements at varied speeds towards a pair of acid-alkali triggers. A series of integrative logic operations and cascaded logic circuits, namely, AND, NAND, NOT, OR, NOR, and NIMPLY, have been constructed employing the decisive chemotactic migrations of the logibot in the presence of the pH gradient established by the sole or coupled effects of acid (HCl-catalase) and alkali (NaOH) drips inside a peroxide bath. The imposed acid and/or alkali triggers across the logibots were realized as inputs while the logic gates were functionally reconfigured to several operational modes by varying the pH of the acid-alkali inputs. The self-propelling logibot could rapidly sense the external stimuli, decide, and act on the basis of intensities of the pH triggers. The impulsive responses of the logibots towards and away from the external acid-alkali stimuli were interpreted as the potential outputs of the logic gates. The external stimuli responsive self-propulsion of the logibots following different logic gates and circuits can not only be an eco-friendly alternative to the silicon-based computing operations but also be a promising strategy for the development of intelligent pH-responsive drug delivery devices.

Many fungi are polykaryotic, containing multiple nuclei per cell. In the case of heterokaryons, there are different nuclear types within a single cell. It is unknown what the different nuclear types contribute in terms of mRNA expression levels in fungal heterokaryons. Each cell of the mushroom Agaricus bisporus contains two to 25 nuclei of two nuclear types originating from two parental strains. Using RNA-sequencing data, we assess the differential mRNA contribution of individual nuclear types and its functional impact. We studied differential expression between genes of the two nuclear types, P1 and P2, throughout mushroom development in various tissue types. P1 and P2 produced specific mRNA profiles that changed through mushroom development. Differential regulation occurred at the gene level, rather than at the locus, chromosomal, or nuclear level. P1 dominated mRNA production throughout development, and P2 showed more differentially up-regulated genes in important functional groups. In the vegetative mycelium, P2 up-regulated almost threefold more metabolism genes and carbohydrate active enzymes (cazymes) than P1, suggesting phenotypic differences in growth. We identified widespread transcriptomic variation between the nuclear types of A. bisporus Our method enables studying nucleus-specific expression, which likely influences the phenotype of a fungus in a polykaryotic stage. Our findings have a wider impact to better understand gene regulation in fungi in a heterokaryotic state. This work provides insight into the transcriptomic variation introduced by genomic nuclear separation., Competing Interests: The authors declare no conflict of interest., (Copyright © 2018 the Author(s). Published by PNAS.)

Fucose is one of important residues of recognition pattern for many immune cells. In this study, we characterized bioactive fucose-containing acidic polysaccharides from submerged fermentation of Agaricus blazei Murill. We obtained the polysaccharides through a cell-based activity-guided strategy, and used carbohydrate recognition monoclonal antibodies based Enzyme-Linked Immuno Sorbent Assay (ELISA) along with methylation and NMR analyses to investigate the structural characteristics of the polysaccharides. The polysaccharides had Mw of 3.5 × 10 5  Da. The major sugars were l-fucose, l-arabinose, d-galactose, d-xylose, and d-galacturonic acid in the molar ratio of 6.4, 15.5, 28.5, 14.7, and 25.0% with a small amount of d-glucose, d-mannose, l-rhamnose, and d-glucuronic acid. Results indicated that the bioactive polysaccharides consisted of a (1,4)-Galp and (1,4)-GalAp back bone; (1,2)-Xyl and (1,2)-Rha might also comprise backbone or constitute side chain; linkage (1,5)-Ara and terminal fucosyl residues were also involved in the polysaccharides. Regarding bioactivity, removal of the terminal l-fucosyl residues reduced the TNF-α cytokine stimulating activity of the polysaccharides in a RAW 264.7 macrophage cell-line test, whereas NF-κB and TLR4 affected the polysaccharide-induced TNF-α production., (Copyright © 2017. Published by Elsevier B.V.)

The white button mushroom Agaricus bisporus is one of the most widely produced edible fungus with a great economical value. Its commercial cultivation process is often performed on wheat straw and animal manure based compost that mainly contains lignocellulosic material as a source of carbon and nutrients for the mushroom production. As a large portion of compost carbohydrates are left unused in the current mushroom cultivation process, the aim of this work was to study wild-type A. bisporus strains for their potential to convert the components that are poorly utilized by the commercial strain A15. We therefore focused our analysis on the stages where the fungus is producing fruiting bodies. Growth profiling was used to identify A. bisporus strains with different abilities to use plant biomass derived polysaccharides, as well as to transport and metabolize the corresponding monomeric sugars. Six wild-type isolates with diverse growth profiles were compared for mushroom production to A15 strain in semi-commercial cultivation conditions. Transcriptome and proteome analyses of the three most interesting wild-type strains and A15 indicated that the unrelated A. bisporus strains degrade and convert plant biomass polymers in a highly similar manner. This was also supported by the chemical content of the compost during the mushroom production process. Our study therefore reveals a highly conserved physiology for unrelated strains of this species during growth in compost., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Solid-state fermentation (SSF) with Agaricus brasiliensis and Agaricus bisporus on corn was carried out. The results showed that SSF with the two fungi made up the deficiency of tryptophan in corn and improved the protein nutritional value of corn. The conjugated polyphenols contents in fermented corn decreased and free polyphenols (FPP) contents increased. FPP contents in corn fermented with the two fungi reached respectively 25 and 88 times of control, total polyohenols contents reached respectively 1.4 and 3.3 times of control. The antioxidant properties (i.e. 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity, reducing power, ferrous ion chelating ability and superoxide anion radical scavenging ability) of fermented corn were improved significantly. SSF with A. bisporus was more favorable to the enhancement in protein nutritional value and antioxidant properties of corn than that of A. brasiliensis. The results indicated that SSF with the two fungi could upgrade the protein nutritional value, FPP content and antioxidant properties of corn.

Substrate preparation (i.e., composting) for Agaricus bisporus cultivation is the most critical point of mushroom production. Among many factors involved in the composting process, the microbial ecology of the system is the underlying drive of composting and can be influenced by composting management techniques. Pasteurization temperature at the beginning of phase II, in theory, may influence the bacterial community and subsequently the "selectivity" and nutrition of the final substrate. Therefore, this hypothesis was tested by simulation in bioreactors under different pasteurization conditions (57 °C/6 h, 60 °C/2 h, and 68 °C/2 h), simulating conditions adopted by many producers. Bacterial diversity, based on 16S ribosomal RNA obtained by high-throughput sequencing and classified in operational taxonomic units (OTUs), was greater than previously reported using culture-dependent methods. Alpha diversity estimators show a lower diversity of OTUs under a high-temperature pasteurization condition. Bacillales order shows a relatively higher OTU abundance under a high-pasteurization temperature, which also was related to high ammonia emission measurements. On the other hand, beta diversity analysis showed no significantly changes in the bacterial community structure under different conditions. Agaricus bisporus mycelium growth during a standard spawn run period was significantly slower in the compost pasteurized at high temperature. Since the bacterial community structure was not greatly affected by different pasteurization conditions but by-products left (e.g., ammonia) at the end of compost conditioning varied, further studies need to be conducted to determine the functional role of the microbial communities found during substrate preparation for Agaricus bisporus cultivation.

An 8-week feeding trial was performed to assess the effects of dietary white button mushroom (Agaricus bisporus) powder (WBMP) on the mucosal immunity and growth of rainbow trout (Oncorhynchus mykiss). Trout (n = 192; weight 13.76 ± 1.17 g) were stocked in 12 cages (65 × 65 × 65 cm) placed in 4 raceways with a flow-through water system. Trout were fed a basal diet (control group) or a basal diet supplemented with 0.5%, 1%, or 2% WBMP for 8 weeks. Evaluation of total protein levels and lysozyme activity in skin mucus revealed noticeable increases in trout fed 1% or 2% WBMP (P < 0.05), but no significant difference was observed with 0.5% WBMP administration (P > 0.05). The results of sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed alterations in the protein profile of skin mucus following dietary administration of WBMP. Molecular studies showed a noticeable increase (P < 0.05) in tumor necrosis factor-α messenger RNA in the intestine of WBMP-fed trout, regardless of the inclusion level. Also, fish receiving the 1% or 2% WBMP treatments had a remarkable increase in interleukin (IL)-1β expression compared with the control group (P < 0.05). In a similar way, intestinal IL-8 expression was upregulated with the 1% and 2% WBMP treatments (P < 0.05). However, no significant difference was found between the control group and the 0.5% WBMP treatment group in the case of IL-8 gene expression (P > 0.05). Furthermore, after 8 weeks of WBMP feeding, no improvement was seen in the growth parameters of trout compared with those fish fed the nonsupplemented diet (P > 0.05). These results hint at the potential immunomodulatory effects of dietary WBMP.

Five culinary-medicinal mushrooms are commonly available in the Malaysian market: Agaricus bisporus (white and brown), Ganoderma lucidum, Hypsizygus marmoreus, Pleurotus floridanus, and P. pulmonarius. These species were selected for use in the current study, the aim of which was to investigate the antimelanogenesis and anti-inflammatory activity of these mushrooms in an attempt to evaluate their potential use in cosmeceuticals. Mushroom fruiting bodies were extracted with hot water, and the extracts were freeze-dried before testing. The antimelanogenesis activity of the extracts was determined by cell viability assay, measurement of intracellular melanin content, and cellular tyrosinase assay with B16F10 melanoma cells. The anti-inflammatory activity of the mushroom extracts was tested by measuring the levels of nitric oxide (NO), tumor necrosis factor (TNF)-α, and interleukin-10 excreted by RAW264.7 macrophages. Brown A. bisporus reduced intracellular melanin content to the largest extent-up to 57.05 ± 3.90%-without a cytotoxic effect on B16F10 melanoma cells. This extract also reduced cellular tyrosinase activity to 17.93 ± 2.65%, performing better than kojic acid, the positive control. In parallel, the extract from brown A. bisporus, at the highest concentration tested, has appreciable anti-inflammatory activity through reductions of NO and TNF-α levels. The other 5 extracts showed moderate antimelanogenesis and anti-inflammatory activities. In summary, our findings show that A. bisporus (brown) extract has the potential to be used as an ingredient in whitening skincare products and to sooth the inflammatory response on the skin.

Providing guidance on the reasonable use of pesticide in agricultural production is of particular importance for ensuring food safety. In the present study, a field trial was performed to study the dissipation and accumulative pattern of cyromazine (CA) and its metabolite in Agaricus bisporus (A. bisporus) cultivation. An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was first developed and validated for the determination of CA and melamine (MEL) in the casing soil and fruiting body. During the cultivation period, the dissipation rates of CA in the casing soil were between 51.57 and 63.48% at three dose groups. The fruiting body presented higher accumulation ability for MEL compared with CA. The terminal residues of MEL never exceeded the maximum residue limits (MRLs) in food. In addition, the intake health risk from the CA and MEL residues in the fruiting body were negligible to humans. This study will help to provide valuable guidance on the application strategies of CA in A. bisporus cultivation.

We previously showed that luminous fungi share a common mechanism in bioluminescence, and identified hispidin as a luciferin precursor in Neonothopanus nambi mycelium. Here we showed the presence of hispidin as a bioluminescent active compound at 25-1000 pmol g -1 in the fruiting bodies of Mycena chlorophos, Omphalotus japonicus, and Neonothopanus gardneri. These results suggest that luminous mushrooms contain hispidin as a luciferin precursor. We also found that non-luminous "young" fruiting bodies exhibited luminescence by hispidin treatment. Furthermore, we observed a gradual luminescence enhancement of the cell-free fruiting body extract by the addition of hispidin biosynthetic components, namely caffeic acid, ATP and malonyl-CoA. These findings suggest that continuous weak glow of luminous mushrooms is regulated by slow recycling biosynthesis of hispidin.

The goal of this study was to evaluate cadmium and lead accumulation ability of in vitro cultures biomass containing selected edible mushroom species derived from the environment (Laetiporus sulphureus, Imleria badia) and those of commercial origin (Agaricus bisporus). Atomic absorption spectrometry was used to evaluate the content of Cd(II) and Pb(II) on the medium supplemented with Cd(II) or Pb(II), each of them at the same concentration of 5·10 -5 M. The highest concentration of Cd(II) ions was determined in the biomass from L. sulphureus in vitro cultures, while the highest concentration of Pb(II) ions was found in the biomass from A. bisporus in vitro cultures. The greatest Cd(II) and Pb(II) accumulation ability in mycelium per dry weight was shown for L. sulphureus. Among the test species, biomass of A. bisporus showed the lowest ability for the bioaccumulation of Cd(II); however, comparable ability for the remediation of Pb(II) was provided by the biomasses from A. bisporus and I. badia in vitro cultures. The results confirm the possibility of using these mushroom species for remediation and indicate the relationship between bioaccumulation of heavy metals and the test species.

Mushroom protein isolate (MPI) from Agaricus bisporus was hydrolyzed using Alcalase, Pancreatin, Flavourzyme, Alcalase-Pancreatin and Alcalase-Flavourzyme. The obtained hydrolysates (MPHs) were ultrafiltered to generate peptide fractions (UFs) of molecular sizes (<1, 1-3, 3-5 and 5-10kDa). The electrophoretic profile results indicated that the enzymatic systems were efficient in hydrolyzing the MPI into low molecular weight peptides. Hydrolysate yields of >57% and protein recoveries of >43% were obtained. Effective concentration that scavenged 50% (EC 50 ) of DPPH radicals was similar for the MPHs while inhibition against linoleic acid oxidation was strongest (66.49%) for Alcalase-Flavourzyme hydrolysate on day 5 of incubation. UFs exhibited a concentration-dependent FRAP, with the highest activity for fractions from Alcalase and Pancreatin recorded in 1-3kDa. The antioxidant activities of MPHs and their UFs suggested that they could be potential bioactive ingredients for use in the formulation of functional foods as well as natural antioxidants in lipid food systems., (Copyright © 2017. Published by Elsevier Ltd.)

The present work is aimed at developing nutraceutical formulations based on the mycelium of Agaricus bisporus and Pleurotus ostreatus, highlighting the potential of in vitro culture as a tool to improve the production of bioactive compounds, namely phenolic acids and ergosterol. The mycelia of both species were cultured in different solid and liquid media in order to compare the growth rate and yielded biomass. Fruiting bodies, mycelia and culture media were compared regarding the antioxidant activity, anti-inflammatory effects in RAW264.7 cells and cytotoxicity in human tumor cell lines and non-tumor porcine liver cells. P. ostreatus mycelia showed higher contents of ergosterol and phenolic compounds, and stronger antioxidant activity than the corresponding fruiting body. P. ostreatus and A. bisporus did not show anti-inflammatory activity, and P. ostreatus was the only one showing cytotoxicity in tumor cell lines. The results show that these mushrooms provide compounds with antioxidant and cytotoxic capacities, with variations among species.

The production of a protease inhibitor from Agaricus bisporus through solid-state fermentation was studied. The purpose was to produce protease inhibitor from natural, cheap, and readily available carbon and nitrogen sources. Solid-state fermentation enhanced the mycelia growth and also gave a higher yield of the product. Further, fungal growth and other production parameters were statistically optimized. The specificity of the inhibitor was tested and was effective against trypsin. Screening of significant factors (wheat bran, cyanobacterial biomass, initial pH, temperature, incubation period, and moisture content and inoculum size) was performed using Plackett-Burman design. Central composite design was used to determine the optimized values of the significant variables which were found to be temperature (27.5°C), incubation time (156 hr), cyanobacterial biomass (1 g), and moisture content (50%) and gave a statistical yield of 980 PIU/g which was 25.6% higher than experimental yield (780 PIU/g). The inhibitor was purified by ammonium sulfate precipitation and diethylaminoethyl (DEAE) cellulose chromatography (yield 43.89% and 0.21%, respectively) and subjected to reversed-phase HPLC to validate its identity. Since protease inhibitors act against proteases, finding ample therapeutic roles; the isolated protease inhibitor from A. bisporus can also be a probable medicinal agent after its further characterization.

Agaricus bisporus (J. E. Lange) Imbach, well known as the common cultivated mushroom or white button mushroom, is widely cultivated all over the world. We used iTRAQ-MS/MS (i.e., isobaric tags for relative and absolute quantification-coupled two-dimensional liquid chromatography-tandem mass spectrometry) to identify protein expression changes that occur during the post-harvest maturation of Agaricus bisporus fruitbodies at 0, 6, 12, and 48 h after harvest. A total of 5878 unique peptides, representing 1063 proteins, were identified. Quantitative data were obtained from 1012 out of the 1063 proteins. A total of 102, 106, and 160 differentially expressed proteins (>twofolds differences) were identified from samples collected at 6, 12, and 48 h compared to sample from 0 h post harvest, accounting for 10.1, 10.5, and 15.8% of the total proteins identified, respectively. All identified proteins including the differentially expressed proteins among different post-harvest stages were subjected to bioinformatic analysis. Furthermore, seven representative proteins either up or down-regulated at different post-harvest stages were analyzed by real-time PCR. Three out of the seven proteins, the mismatched base pair and cruciform DNA recognition protein, hydrophobin-B and protein transporter, exhibited similar expression patterns as their corresponding proteins. The 260 differentially expressed proteins identified from our study laid a foundation for future studies aiming to understand the post-harvest maturation process of A. bisporus and eventually, might help in the development of breeding program to identify strains with extended shelf life.

In order to study the respiration metabolism mechanism based on the generation of adenosine triphosphate (ATP) and reactive oxygen species (ROS) and nitric oxide (NO) by the electron transport chain (ETC) of the white mushroom under high O 2 /CO 2 controlled atmospheres, the treatments of 100% O 2 , 80% O 2 + 20% CO 2 , 60% O 2 + 40% CO 2 , and 40% O 2 + 60% CO 2 at 2 ± 1 °C were employed and natural air was used as the control. ATP and energy charge can maintain the membrane integrity and function, life activities, and physicochemical reactions of higher plants. The results showed that the 80% O 2 + 20% CO 2 treatment inhibited the respiration rate, embden-meyerhof-parnas or glycolysis pathway, and ROS and NO contents. It significantly delayed the reduction of the ATP content and energy charge level, tricarboxyfic-acid-cycle and cytochrome pathway proportion, and their key enzymes activity and gene expression. It also maintained a high phosphopentose pathway and moderate alternative pathway. Results indicated that the 80% O 2 + 20% CO 2 prolonged the storage time of mushrooms to 24 days and retarded the senescence through retaining the higher energy, suppressing the ROS contents, enhancing the endurance capability in adversity, and regulating the respiration pathways and ETC metabolism.

Multigenicity is commonly found in fungal enzyme systems, with the purpose of functional compensation upon deficiency of one of its members or leading to enzyme isoforms with new functionalities through gene diversification. Three genes of the flavin-dependent glucose-methanol-choline (GMC) oxidoreductase pyranose dehydrogenase (AmPDH) were previously identified in the litter-degrading fungus Agaricus (Leucoagaricus) meleagris, of which only AmPDH1 was successfully expressed and characterized. The aim of this work was to study the biophysical and biochemical properties of AmPDH2 and AmPDH3 and compare them with those of AmPDH1. AmPDH1, AmPDH2 and AmPDH3 showed negligible oxygen reactivity and possess a covalently tethered FAD cofactor. All three isoforms can oxidise a range of different monosaccarides and oligosaccharides including glucose, mannose, galactose and xylose, which are the main constituent sugars of cellulose and hemicelluloses, and judging from the apparent steady-state kinetics determined for these sugars, the three isoforms do not show significant differences pertaining to their reaction with sugar substrates. They oxidize glucose both at C2 and C3 and upon prolonged reaction C2 and C3 double-oxidized glucose is obtained, confirming that the A. meleagris genes pdh2 (AY753308.1) and pdh3 (DQ117577.1) indeed encode CAZy class AA3&#95;2 pyranose dehydrogenases. While reactivity with electron donor substrates was comparable for the three AmPDH isoforms, their kinetic properties differed significantly for the model electron acceptor substrates tested, a radical (the 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulphonic acid] cation radical), a quinone (benzoquinone) and a complexed iron ion (the ferricenium ion). Thus, a possible explanation for this PDH multiplicity in A. meleagris could be that different isoforms react preferentially with structurally different electron acceptors in vivo.

Objective: We studied the dynamic nuclear behavior of Hypomyces perniciosus on axenic culture and its disease progression after infection on different growth stages of Agaricus bisporus., Methods: Infection process was initiated by inoculating different stages of A. bisporus fruit body, and different depths of compost and casing soil with H. perniciosus. Disease progression was studied by observing symptoms on the fruit body using light microscopy and scanning electron microscopy. The nuclear behavior of H. perniciosus was determined by observation using fluorescence light microscopy after binding of DNA specific fluorochrome dye (DAPI:4, 6-Diamidino-2-phenylindole dihydrochloride) to the nuclei., Results: Inoculating H. perniciosus on different depths of compost and casing soil resulted in different disease rate as follows:on the surface of casing soil>in the center of casing soil>between the casing soil and the compost>in the center of compost. H. perniciosus can infect any stage of fruit body development, when young primordial (up to 3 mm) was infected, large, irregular and tumorous fungal masses were formed. H. perniciosus directly penetrated A. bisporus without the formation of appressorium-like structures. The germination of the conidia led to a necrotic brown lesion symptom on A. bisporus at the beginning stages of disease development. The mycelium of A. bisporus plasmolysed, hydropically degenerated, cytoplasmolysed, emptied of mycelium cytosol and eventual death as the disease advanced. H. perniciosus produced two types of conidia. Group I conidia had no septa, colorless and smooth containing one nucleus. Group II didymoconidium had septa, containing two nuclei, separated by septa. The first round of mitosis occurred in conidia with no nucleus in the germinal tube. Another kind of asexual spore for thicker cell wall wart convex chlamydospore, chlamydospore had two cells. The upper cell had two nuclear while the basal cell had one or two nuclear, when germinated, it produced one or two germinal tubes. The number of nuclear in the germinal tube was irregular, usually contained 0 to 2 nuclear., Conclusion: H. perniciosus can infect any part of the A. bisporus fruit body and can cause tremendous cytology changed. If we perform single spore isolation to do genetic analysis, one must isolate conidia with no septa.

Background: Agaricus bisporus (white button mushroom) is an important culinary and medicinal species of worldwide importance. The present study investigated for the first time whether it may be grown on substrates supplemented with Se alone or in combination with Cu and/or Zn (0.1-0.8 mmol L -1 ) to produce fruiting bodies of increased nutritional value., Results: As found, substrate supplementation did not affect yielded biomass up to 0.6 mmol L -1 element concentrations regardless of the cultivation model. At 0.8 mmol L -1 Se + Cu and Se + Zn supplementation biomass comparable with controls still developed. The accumulation of trace elements in the fruiting bodies generally increased over the concentration gradient reaching its maximum at 0.6 mmol L -1 (for Se + Zn and Se + Cu + Zn) and 0.8 mmol L -1 (for Se and Se + Cu). The organic Se constituted the greatest share in total Se quota. As calculated, each 10 g of dried fruiting bodies of A. bisporus obtained from 0.6 or 0.8 mmol L -1 supplementation would represent 342-469% of the Recommended Daily Allowance (RDA) for Se, 43.4-48.5% for Cu and 5.2-5.8% for Zn., Conclusion: Considering inexpensive methods of A. bisporus cultivation, global popularity and use of this mushroom, its biofortification with Se, Cu and Zn could have a practical application in deficiency prevention and assisted treatment. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

This study investigated changes in the activity, conformation and microstructure of mushroom polyphenoloxidase (PPO) subjected to thermal treatment. The inactivation of PPO can be achieved by high temperature-short time or mild temperature-long time treatment. Circular dichroism and fluorescence spectra suggested that heating process induced the rearrangement of secondary structure and the disruption of tertiary structure. Red shifts of fluorescence spectra showed positive correlations with the inactivation rate of PPO. There were significant differences in the conformation and molecular microstructure among PPO samples with the same relative activity, which were obtained by treating PPO at 45, 55 and 65°C for different times. In summary, PPO molecules were deformed at mild temperature, while higher temperature induced the formation of large aggregates. PPO with the same relative activity might exist in different forms., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The ability of Agaricus blazei mushroom in its dried and powdered mycelial form was evaluated for its antigenotoxic properties for the first time. Antigenotoxic effects in human peripheral blood cells against H 2 O 2 -induced DNA damage were examined in pretreatment and posttreatment protocol by comet assay. The results showed better antigenotoxic properties of Agaricus blazei on the interventional level, respectively, after treatment. Agaricus blazei in concentration of 250  μ g/mL after treatment was most efficient in regard to its action against DNA damage. The evaluation of repair kinetics showed decrease in H 2 O 2 induced DNA damage 15 min after the application of A. blazei , reaching the maximum potency after 30 min. Analysis of antioxidant properties of Agaricus blazei revealed strong • OH scavenging properties and moderate reducing power, while its DPPH scavenging ability was weak. In regard to our findings, we can conclude that our preliminary results demonstrated antigenotoxic properties of Agaricus blazei and its strong • OH scavenging ability. Mechanisms underlying its properties should be further evaluated in in vivo studies., Competing Interests: The authors declare that there is no conflict of interests regarding the publication of this paper.

Drought is a stressor for many soil-inhabiting organisms. Although plants have been extensively investigated for drought-adaptive mechanisms, little information is available for fungi. Antioxidants are especially relevant, since desiccation is accompanied by an excessive intracellular production of reactive oxygen species. Riboflavin (vitamin B 2 ) is one antioxidant regulating drought tolerance in plants. A similar function may exist in fungi. Here, we examined the respiratory and transcriptional responses of Agaricus bisporus to drought and the impact of riboflavin. Mesocosm experiments with four groups were established: hyphae were treated with or without 50 µM riboflavin under drought or no drought conditions. Drought increased riboflavin content in hyphae about 5 times with, but also without, addition of riboflavin. Without addition of riboflavin, fungal respiration decreased by more than 50% at water potentials of about -20 MPa. With addition of riboflavin, respiration remained about 2-3 times higher. The transcriptional responses to only drought or only riboflavin strongly overlapped and were mainly based on factors regulating transcription and translation. This was even stronger in combined treatments. Riboflavin induced protective mechanisms in drought-stressed hyphae. Most pronounced was the methylglyoxal (cytotoxic by-product of glycolysis) detoxifying of lactoylglutathione lyase. Thus, our data suggest a stress-priming function and a role of riboflavin in drought responses of A. bisporus.

Background: The novel A/J Min/+ mouse, which is a model for human Familial Adenomatous Polyposis (FAP), develops spontaneously multiple adenocarcinomas in the colon as well as in the small intestine. Agaricus blazei Murill (AbM) is an edible Basidiomycetes mushroom that has been used in traditional medicine against cancer and other diseases. The mushroom contains immunomodulating β-glucans and is shown to have antitumor effects in murine cancer models. Andosan™ is a water extract based on AbM (82%), but it also contains the medicinal Basidiomycetes mushrooms Hericeum erinaceus and Grifola frondosa., Methods and Findings: Tap water with 10% Andosan™ was provided as the only drinking water for 15 or 22 weeks to A/J Min/+ mice and A/J wild-type mice (one single-nucleotide polymorphism (SNP) difference), which then were exsanguinated and their intestines preserved in formaldehyde and the serum frozen. The intestines were examined blindly by microscopy and also stained for the tumor-associated protease, legumain. Serum cytokines (pro- and anti-inflammatory, Th1-, Th2 -and Th17 type) were measured by Luminex multiplex analysis. Andosan™ treated A/J Min/+ mice had a significantly lower number of adenocarcinomas in the intestines, as well as a 60% significantly reduced intestinal tumor load (number of tumors x size) compared to control. There was also reduced legumain expression in intestines from Andosan™ treated animals. Moreover, Andosan™ had a significant cytotoxic effect correlating with apoptosis on the human cancer colon cell line, Caco-2, in vitro. When examining serum from both A/J Min/+ and wild type mice, there was a significant increase in anti-tumor Th1 type and pro-inflammatory cytokines in the Andosan™ treated mice., Conclusions: The results from this mouse model for colorectal cancer shows significant protection of orally administered Andosan™ against development of intestinal cancer. This is supported by the finding of less legumain in intestines of Andosan™ treated mice and increased systemic Th1 cytokine response. The mechanism is probably both immuno-modulatory and growth inhibition of tumor cells by induction of apoptosis., Competing Interests: Author GH has patent/patent applications and financial interests as shareholder in Immunopharma AS of Norway, commercializing material (AndoSan™) pertinent to this article: i) WO2005065063 A2, Appl. No.:10/585600, NO- and PCT-filed Jan 2004 and Jan 2005, respectively, by Inventor Hetland Geir, and ii) NO20090003383, Appl. No.: NO20090003383 20091119, by Inventor Hetland Geir filed by Applicant Immunopharma AS in Nov 2009. This does not alter the authors’ adherence to PLOS ONE policies on sharing data and materials.

Tyrosinase is an oxidase that is the rate-limiting enzyme for controlling the production of melanin in the human body. Overproduction of melanin could lead to a variety of skin disorders. Glabridin, an isoflavan, isolated from the root of Glycyrrhiza glabra Linn, has exhibited several pharmacological activities, including excellent inhibitory effects on tyrosinase. In this paper, the inhibitory kinetics of glabridin on tyrosinase and their binding mechanisms were determined using spectroscopic, zebrafish model and molecular docking techniques. The results indicate that glabridin reversibly inhibits tyrosinase in a noncompetitive manner through a multiphase kinetic process with the IC50 of 0.43μmol/L. It has been shown that glabridin had a strong ability to quench the intrinsic fluorescence of tyrosinase mainly through a static quenching procedure, suggesting a stable glabridin-tyrosinase complex may be generated. The results of molecular docking suggest that glabridin did not directly bind to the active site of tyrosinase. Moreover, according to the results of zebrafish model system, glabridin shows no effects on melanin synthesis in zebrafish but presents toxicity to zebrafish embryo. The possible inhibitory mechanisms, which will help to design and search for tyrosinase inhibitors especially for glabridin analogues, were proposed., (Copyright © 2016. Published by Elsevier B.V.)

Background: This work assesses the agronomic performance of defatted pistachio meal, after oil extraction, as a nutritional substrate supplement when growing the mushroom species Agaricus bisporus (Lange) Imbach and Pleurotus ostreatus (Jacq.) P. Kumm. Materials were applied at different doses at spawning. Along with non-supplemented substrates, commercial nutritional supplements were used as controls. Proximate analysis of mushrooms is also considered., Results: For the cultivation of champignon, defatted pistachio meal has provided larger mushrooms (unitary weight and cap diameter) with firmer texture and greater content in dry weight and protein, without significant alterations in quantitative parameters. For Pleurotus ostreatus, the supplement led to significant yield increase, even providing up to 34.4% of increment compared to non-supplementation with meal, reaching a biological efficiency of 129.9 kg dt(-1) , when applied to the 15 g kg(-1) compost dose. Supplementation has also been conducted to increase dry weight, protein and fibre within carpophores and to decrease the energy value. Defatted pistachio meal has similar or better results compared to the commercial supplements used as reference., Conclusions: Compost supplementation with defatted pistachio meal in A. bisporus concerns mainly the quantitative parameters (size, texture, dry weight and protein). Based on the results obtained, this technique has greater potential of development for P. ostreatus commercial crops, basically due to expected increases in production, with a direct impact on benefits and crop profitability. © 2015 Society of Chemical Industry., (© 2015 Society of Chemical Industry.)

Ethylene biosynthesis and function in Agaricus bisporus (the button mushroom) remain uncertain. The enzyme activities of 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) and ACC oxidase (ACO) were detectable in A. bisporus AS2796 and inhibited by α-aminooxyacetic acid and Co2+. We cloned and sequenced 2 ACS genes (Ab-ACS1 and Ab-ACS2) and 1 ACO gene (Ab-ACO) from the mushroom strain. Ab-ACS1 and Ab-ACS2 demonstrated low amino acid sequence similarity. Ab-ACO demonstrated an amino acid sequence completely identical to that of ACO1&#95;AGABI from A. bisporus. Antisense ACO significantly reduced ACO gene expression level, ACO enzyme activity, and ethylene production in the mushroom transformants. The transformants grew faster than the wild-type strain in sterilized compost and normally formed primordia when cultivated in sterilized compost with the sterilized casing vermiculite, but the wild-type strain did not. Our results show that ethylene is synthesized in button mushrooms via the ACC pathway. Ethylene inhibited button mushroom mycelial growth and development.

Zinc is one of those microelements that are essential for the proper functioning of the human body and must be supplemented in our food at a daily dose of 15 mg. It is well known that mushrooms accumulate elements; thus, in order to determine the extent of accumulation and the level of zinc released from mushrooms, in vitro cultures of Agaricus bisporus were established. The cultures were run on a modified Oddoux medium (a control culture) as well as on the same medium with the addition of zinc hydroaspartate (100 and 200 mg/L) and zinc sulfate (87.23 and 174.47 mg/L). These compounds were chosen to help estimate which form, organic or inorganic, results in a better assimilation of zinc(II) ions by biomass. As the next step, the level of zinc(II) ions released from the lyophilized biomass of in vitro cultures to the digestive juices, under thermal conditions of the human body (37°C), was determined. For this purpose, artificial digestive juices, imitating the composition of human digestive juices, were used. For determination of zinc(II) ions in the digestive tract, an anodic stripping voltammetry method was employed. The amount of zinc released into artificial saliva over 1 minute varied from 0.15 mg/100 g d.w. in the control culture to 2.35 mg/100 g d.w. in the biomass in the medium to which 200 mg/L zinc hydroaspartate had been added. Values were higher in gastric juice and depended on incubation time (2.66 to 30.63 mg/100 g d.w.). In intestinal juice, the highest value of the released zinc grew to 24.20 mg/100 g d.w. (biomass of A. bisporus in vitro cultures in medium with the addition of 200 mg/L zinc hydroaspartate). Total average amount of zinc released into artificial digestive juices was the highest (56.26 mg/100 g d.w.) from A. bisporus biomass of in vitro cultures in the medium to which 200 mg/L zinc hydroaspartate had been added.

This study investigates the effect of three spent Agaricus bisporus substrate (SAS) application methods on bioremediation of soil multi-polluted with Pb and PAH from close to a shooting range with respect natural attenuation (SM). The remediation treatments involve (i) use of sterilized SAS to biostimulate the inherent soil microbiota (SSAS) and two bioaugmentation possibilities (ii) its use without previous treatment to inoculate A. bisporus and inherent microbiota (SAS) or (iii) SAS sterilization and further A. bisporus re-inoculation (Abisp). The efficiency of each bioremediation microcosm was evaluated by: fungal activity, heterotrophic and PAH-degrading bacterial population, PAH removal, Pb mobility and soil eco-toxicity. Biostimulation of the native soil microbiology (SSAS) achieved similar levels of PAH biodegradation as SM and poor soil detoxification. Bioaugmented microcosms produced higher PAH removal and eco-toxicity reduction via different routes. SAS increased the PAH-degrading bacterial population, but lowered fungal activity. Abisp was a good inoculum carrier for A. bisporus exhibiting high levels of ligninolytic activity, the total and PAH-degrading bacteria population increased with incubation time. The three SAS applications produced slight Pb mobilization (<0.3%). SAS sterilization and further A. bisporus re-inoculation (Abisp) proved the best application method to remove PAH, mainly BaP, and detoxify the multi-polluted soil., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Laccases are polyphenol oxidases produced by many fungi and have many applications in textile, food and beverage, and pulp and paper industries. Laccase production can be induced using aromatic or phenolic compounds that mostly affect the transcription of laccase-encoding genes. In this study, we analyzed laccase and biomass production by Agaricus blazei in the presence of different concentrations of nitrogen, copper, and inducers such as pyrogallol, veratryl alcohol, xylidine, vanillin, guaiacol, and ethanol. Laccase production by A. blazei U2-4 reached 43.8 U/mL in the presence of 2.8 g/L nitrogen and 150 μM copper. However, addition of copper to the cultivation medium decreased biomass production. Different compounds differentially induced laccase production by A. blazei. Moreover, different concentrations of these inducers exerted different effects on laccase activity. Ethanol (1.0 mM), guaiacol (0.5 mM), and vanillin (0.5 mM) were the best inducers and increased laccase activity by 120% (A. blazei U2-2), 30% (A. blazei U2-3), and 9% (A. blazei U2-4), respectively. In contrast, pyrogallol and xylidine decreased laccase activity but increased biomass production.

Mushroom Virus X (MVX) is associated with a range of symptoms observed in mushroom crops. The most prominent symptom in Ireland is the occurrence of 'brown' or 'off-white' mushrooms in white strain crops. The browning symptoms are associated with the presence of four low molecular weight dsRNAs: MVX(0.6), MVX(0.8), MVX(1.8) and MVX(2.0), however viral dsRNAs also occur in non-symptomatic mushrooms. Three virus-infected mushroom cultures containing MVX(1.8) and MVX(2.0) were used to infect experimental crops at different rates and at different times in the crop cycle to test the effect on symptom expression. Mushroom colour was measured by chromometer, and the ΔE value calculated. RT-PCR was used to test for the presence of MVX(1.8) dsRNA in harvested mushrooms. Results indicate that following infection, browning symptom expression is variable both within and between crops. Control mushrooms from 1st and 2nd flush had ΔE values of 7-12, with most being <10. In contrast, 1st flush mushrooms from virus infected treatments had ΔE values of 6-25, with most being >10 while 2nd flush mushrooms had ΔE values similar to controls. Only mushrooms with ΔE > 15 appeared visibly brown or off colour. The transient and inconsistent nature of MVX-associated browning symptoms is discussed., (Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

The desiccation of upper soil horizons is a common phenomenon, leading to a decrease in soil microbial activity and mineralization. Recent studies have shown that fungal communities and fungal-based food webs are less sensitive and better adapted to soil desiccation than bacterial-based food webs. One reason for a better fungal adaptation to soil desiccation may be hydraulic redistribution of water by mycelia networks. Here we show that a saprotrophic fungus (Agaricus bisporus) redistributes water from moist (-0.03 MPa) into dry (-9.5 MPa) soil at about 0.3 cm ⋅ min(-1) in single hyphae, resulting in an increase in soil water potential after 72 h. The increase in soil moisture by hydraulic redistribution significantly enhanced carbon mineralization by 2,800% and enzymatic activity by 250-350% in the previously dry soil compartment within 168 h. Our results demonstrate that hydraulic redistribution can partly compensate water deficiency if water is available in other zones of the mycelia network. Hydraulic redistribution is likely one of the mechanisms behind higher drought resistance of soil fungi compared with bacteria. Moreover, hydraulic redistribution by saprotrophic fungi is an underrated pathway of water transport in soils and may lead to a transfer of water to zones of high fungal activity.

Laccases are used by fungi for several functions including defence responses to stresses associated with attack by other fungi. Laccase activity changes and the induction of two laccase genes, lcc1 and lcc2, in Agaricus bisporus were measured in response to toxic extracts of medium in which Trichoderma aggressivum, the cause of green mould disease, was grown. A strain of A. bisporus that shows resistance to the extracts showed higher basal levels and greater enzymatic activity after extract exposure than did a sensitive strain. Furthermore, pre-incubation of T. aggressivum extract with laccases reduced toxicity. Faster induction and greater numbers of lcc2 transcripts in response to the extract were noted in the resistant strain than in the sensitive strain. The timing and increase in lcc2 transcript abundance mirrored changes in total laccase activity. No correlation between resistance and lcc1 transcription was apparent. Transcript abundance in transformants with a siRNA construct homologous to both genes varied widely. A strong negative correlation between transcript abundance and sensitivity of the transformant to toxic extract was observed in plate assays. These results indicated that laccase activity and in particular that encoded by lcc2 contributes to toxin metabolism and by extension green mould disease resistance., (© 2015 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.)

In wheat straw based composting, enabling growth of Agaricus bisporus mushrooms, it is unknown to which extent the carbohydrate-lignin matrix changes and how much is metabolized. In this paper we report yields and remaining structures of the major components. During the Phase II of composting 50% of both xylan and cellulose were metabolized by microbial activity, while lignin structures were unaltered. During A. bisporus' mycelium growth (Phase III) carbohydrates were only slightly consumed and xylan was found to be partially degraded. At the same time, lignin was metabolized for 45% based on pyrolysis GC/MS. Remaining lignin was found to be modified by an increase in the ratio of syringyl (S) to guaiacyl (G) units from 0.5 to 0.7 during mycelium growth, while fewer decorations on the phenolic skeleton of both S and G units remained.

The dose response effect of UV-B irradiation from sunlight on vitamin D2 content of sliced Agaricus bisporus (white button mushroom) during the process of sun-drying was investigated.Real-time UV-B and UV-A data were obtained using a high-performance spectroradiometer. During the first hour of sunlight exposure, the vitamin D2 content of the mushrooms increased in a linear manner, with concentrations increasing from 0.1 μg/g up to 3.9 ± 0.8 μg/g dry weight (DW). At the subsequent two measurements one and 3 h later, respectively, a plateau was reached. Two hours of additional exposure triggered a significant decline in vitamin D2 content. After just 15 min of sun exposure and an UV-B dose of 0.13 J/cm(2), the vitamin D2 content increased significantly to 2.2 ± 0.5 μg/g DW (P < 0.0001), which is equivalent to 17.6 μg (704 IU) vitamin D2 per 100 g of fresh mushrooms and comparable to levels found in fatty fish like the Atlantic salmon.

The economically important edible basidiomycete mushroom Agaricus bisporus thrives on decaying plant material in forests and grasslands of North America and Europe. It degrades forest litter and contributes to global carbon recycling, depolymerizing (hemi-)cellulose and lignin in plant biomass. Relatively little is known about how A. bisporus grows in the controlled environment in commercial production facilities and utilizes its substrate. Using transcriptomics and proteomics, we showed that changes in plant biomass degradation by A. bisporus occur throughout its life cycle. Ligninolytic genes were only highly expressed during the spawning stage day 16. In contrast, (hemi-)cellulolytic genes were highly expressed at the first flush, whereas low expression was observed at the second flush. The essential role for many highly expressed plant biomass degrading genes was supported by exo-proteome analysis. Our data also support a model of sequential lignocellulose degradation by wood-decaying fungi proposed in previous studies, concluding that lignin is degraded at the initial stage of growth in compost and is not modified after the spawning stage. The observed differences in gene expression involved in (hemi-)cellulose degradation between the first and second flushes could partially explain the reduction in the number of mushrooms during the second flush., (© 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.)

The white button mushroom, Agaricus bisporus, is the most commonly grown mushroom in Iran; however, there is a significant shortage of research on its antioxidant activity and other medicinal properties. The aim of this study was to evaluate antioxidant capacity of the methanolic extracts from four cultivated strains and four Internal Transcribed Spacer (ITS)-identified, Iranian wild isolates of A. bisporus. Evaluations were made for total phenols, flavonoids and anthocyanins, and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. Overall, results showed that all the wild isolates exhibited significantly lower DPPH-derived EC50, compared to the cultivated strains (p < 0.05). A relatively high relationship was observed between total phenols and flavonoids or anthocyanins (r(2) > 0.60). However, these constituents could not statistically differentiate the group of wild samples from the cultivated ones, and there was low correlation with the DPPH-derived EC50s (r(2) < 0.40). In conclusion, comparisons showed that wild isolate 4 and cultivated strains A15 and H1 had higher antioxidant capacity than the others (p < 0.05). This result identifies these mushrooms as good candidates for further investigation.

Different applications of spent Agaricus bisporus substrate (SAS), a widespread agro-industrial waste, were investigated with respect to the remediation of a historically polluted soil with Polycyclic Aromatic Hydrocarbons (PAH). In one treatment, the waste was sterilized (SSAS) prior to its application in order to assess its ability to biostimulate, as an organic amendment, the resident soil microbiota and ensuing contaminant degradation. For the other treatments, two bioaugmentation approaches were investigated; the first involved the use of the waste itself and thus implied the application of A. bisporus and the inherent microbiota of the waste. In the second treatment, SAS was sterilized and inoculated again with the fungus to assess its ability to act as a fungal carrier. All these treatments were compared with natural attenuation in terms of their impact on soil heterotrophic and PAH-degrading bacteria, fungal growth, biodiversity of soil microbiota and ability to affect PAH bioavailability and ensuing degradation and detoxification. Results clearly showed that historically PAH contaminated soil was not amenable to natural attenuation. Conversely, the addition of sterilized spent A. bisporus substrate to the soil stimulated resident soil bacteria with ensuing high removals of 3-ring PAH. Both augmentation treatments were more effective in removing highly condensed PAH, some of which known to possess a significant carcinogenic activity. Regardless of the mode of application, the present results strongly support the adequacy of SAS for environmental remediation purposes and open the way to an attractive recycling option of this waste., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Many species of terrestrial fungi produce fruiting bodies that contain high proportions of arsenobetaine (AB), an arsenic compound of no known toxicity. It is unknown whether fungi produce or accumulate AB from the surrounding environment. The present study targets the vegetative life stage (mycelium) of fungi, to examine the role of this stage in arsenic transformations and potential formation of AB. The mycelia of three different fungi species were cultured axenically and exposed to AB, arsenate (As(V)) and dimethylarsinoyl acetic acid for 60 days. Agaricus bisporus was additionally exposed to hypothesized precursors for AB and the exposure time to As(V) and dimethlyarsinic acid was also extended to 120 days. The mycelia of all fungi species accumulated all arsenic compounds with two species accumulating significantly more AB than other compounds. Few biotransformations were observed in these experiments indicating that it is unlikely that the mycelium of the fungus is responsible for biosynthesizing AB., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Oxidative-nitrative stress develops as a result of hyperglycemia under diabetes mellitus. Formation of excessive reactive oxygen species and reactive nitrogen species leads to different cytotoxic effects and ultimately to increased cell death by apoptosis of immune-competent blood cells. This study showed the influence of medicinal mushroom (MM) administration on the L-arginine/nitric oxide (NO) system and rat leukocyte apoptosis under normal and experimental diabetic conditions. Animals were divided into 6 groups: (1) control, (2) control animals treated with Agaricus brasiliensis, (3) control animals treated with Ganoderma lucidum, (4) animals with experimental diabetes (EDM), (5) diabetic animals treated with A. brasiliensis, and (6) diabetic animals treated with G. lucidum. Control and diabetic animals were fed powdered mushrooms at a dose of 1 g/kg body weight. Administration of MMs to animals with diabetes caused a decrease in the activity of the NO synthase enzyme, as well as in the content of stable end products of NO metabolism-nitrates and nitrites-at the control level. The normalizing effect of mushrooms on the percentage of leukocytes that contain pro- (p53) and antiapoptotic (Bcl-2) proteins compared with the EDM group was shown by immunocytochemical analysis. Thus the administration of MMs under EDM showed a positive corrective action on the L-arginine/NO system and the ratio between p53 and Bcl-2 proteins in white blood cells, as well as on apoptotic index reduction.

Centuries of artificial selection have dramatically improved the yield of human agriculture; however, strong directional selection also occurs in natural symbiotic interactions. Fungus-growing attine ants cultivate basidiomycete fungi for food. One cultivar lineage has evolved inflated hyphal tips (gongylidia) that grow in bundles called staphylae, to specifically feed the ants. Here we show extensive regulation and molecular signals of adaptive evolution in gene trancripts associated with gongylidia biosynthesis, morphogenesis and enzymatic plant cell wall degradation in the leaf-cutting ant cultivar Leucoagaricus gongylophorus. Comparative analysis of staphylae growth morphology and transcriptome-wide expressional and nucleotide divergence indicate that gongylidia provide leaf-cutting ants with essential amino acids and plant-degrading enzymes, and that they may have done so for 20-25 million years without much evolutionary change. These molecular traits and signatures of selection imply that staphylae are highly advanced coevolutionary organs that play pivotal roles in the mutualism between leaf-cutting ants and their fungal cultivars.

The present study aimed to clarify the effects of Agaricus brasiliensis KA21 (i.e., Agaricus blazei) mushroom on circulatory function. Spontaneously hypertensive rats (SHRs) were fed 10% A. blazei-containing pellets (agaricus group) or normal pellets (control group) for 5 weeks from 6 to 11 weeks of age. For Experiment 1, tail blood pressure and heart rate were measured in the conscious SHRs. For Experiment 2, echocardiographic and blood biochemical measurements were performed in the anesthetized SHRs. In Experiment 1, blood pressure and heart rate were significantly lower in the agaricus group compared with the control group throughout the observation period. In Experiment 2, the agaricus group also showed a significant decrease in cardiac output accompanied by a decrease in heart rate and an increase in early and late ventricular filling velocity (E/A ratio). Moreover, levels of escape enzymes such as creatine kinase (CK), CK-BB, CK-MB, asparate aminotransferase, lactate dehydrogenase, and aldolase were significantly lower than in the control group. We concluded that the ingestion of feed containing A. brasiliensis KA21 can improve hypertensive cardiovascular hemodynamics by decreasing the working load of the heart, presumably by lowering the sympathetic nervous tone in SHRs.

The effect of dietary supplementation with Se-enriched Agaricus bisporus on cytosolic gluthathione peroxidase-1 (GPx-1), gastrointestinal specific glutathione peroxidase-2 (GPx-2), thioredoxin reductase-1 (TrxR-1) and selenoprotein P (SeP) mRNA expression and GPx-1 enzyme activity in rat colon was examined. Rats were fed for 5weeks with control diet (0.15μg Se/g feed) or Se-enriched diet fortified with selenised mushroom (1μg Se/g feed). The mRNA expression levels were found to be significantly (P<0.01) up-regulated by 1.65-fold and 2.3-fold for GPx-1 and GPx-2, respectively, but were not significantly different for TrxR-1 and SeP between the 2 diet treatments. The up-regulation of GPx-1 mRNA expression was consistent with GPX-1 activity level, which was significantly (P<0.05) increased by 1.77-fold in rats fed with the Se-enriched diet compared to the control diet. The results showed that selenised A. bisporus can positively increase GPx-1 and GPx-2 gene expression and GPx-1 enzyme activity in rat colon., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)