Bioconversion of agricultural lignocellulosic wastes through the cultivation of the edible mushrooms Agrocybe aegerita,Volvariella volvacea and Pleurotus spp.

Ten selected wild and commercial strains of Pleurotus ostreatus,Pleurotus eryngii,Pleurotus pulmonarius, Agrocybe aegerita andVolvariella volvacea were cultivated on three agricultural wastes, i.e. wheat straw (WS), cotton waste (CW) and peanut shells (PS). All species demonstrated significantly higher colonization rates on WS and CW than on PS. WS supported faster growth of A. aegerita and Pleurotus spp., whereas V. volvacea performed better on CW. Comparison of growth rates on composted and non-composted WS and CW substrates revealed that in the latter case faster colonization was achieved, particularly for Pleurotus spp. However, one commercial strain of V. volvacea presented higher growth rates when the composted CW medium was used. Furthermore, earliness in the fructification of P. ostreatus, P. pulmonarius and V. volvacea strains was promoted in CW substrates, while WS favoured earliness of P. eryngii and A. aegerita. Similarly, high sporophore yields were obtained by P. ostreatus and P. pulmonarius on both wastes, whereas WS enhanced yield and basidioma size of P. eryngii and A. aegerita strains and CW production of V. volvacea. The substrates cellulose:lignin ratios were found to be positively correlated to mycelial growth rates and to mushroom yield of P. ostreatus and P. pulmonarius; in addition, positive correlation was also detected for carbon:nitrogen ratio and mushroom yield in P. eryngii and A. aegerita and between cellulose content and mushroom yield for V. volvacea strains.     

Response of rhizosphere microbial diversity and soil physico-chemical properties in a rotation of cucumber with Volvariella volvacea

Crop rotation can significantly increase yield by improving soil conditions through reducing soil pathogens and increasing the microbial diversity near the root system of the plant. Cucumber is widely consumed across the world, and monoculture of cucumber under greenhouse conditions is often threatened by Fusarium wilt, which is caused by fungi belonging to the genus Fusarium. In Jiangsu province of China, rotating cucumber crops with Volvariella volvacea is used as a way to control Fusarium wilt. Here, we tested how rotating cucumber with V. volvacea influences soil physico-chemical and biological properties. We found that rotating with V. volvacea improved bacterial diversity near the cucumber root system. We also found an increase in nitrogen-fixing bacteria that have been used for biological control of plant pathogens, and a decrease in Fusarium species. In the treated soil, we found increased activity of the soil enzymes catalase, dehydrogenase, polyphenol oxidase and alkaline phosphatase by 2–22%, 14–29%, 16–58% and 9–64%, respectively. Lastly, the total and available nitrogen, phosphorus and potassium content significantly increased one to three weeks after V. volvacea residual substrate was mixed into the soil, up to 39%, 24%, 17%, 28%, 18% and 70, but declined slowly afterwards. These results suggested that rotating cucumber with V. volvacea could aid in resetting the structure of the microbial community of the cucumber root system. Increase in beneficial microbes and improved soil conditions are likely to contribute to improved yield of this commercially important crop.     

Lignocellulolytic enzyme profiles of edible mushroom fungi

One of the most economically-viable processes for the bioconversion of many types of lignocellulosic wastes is represented by edible mushroom cultivation. Lentinula edodes, Volvariella volvacea and Pleurotus sajor-caju are three important commercially cultivated mushrooms which exhibit varying abilities to utilise different lignocellulosics as growth substrate. Examination of the lignocellulolytic enzyme profiles of the three species show this diversity to be reflected in qualitative variations in the major enzymic determinants (i.e. cellulases, ligninases) required for substrate bioconversion. For example, L. edodes, which is cultivated on highly lignified substrates such as wood or sawdust, produces two extracellular enzymes which have been associated with lignin depolymerisation in other fungi, (manganese peroxidase and laccase). Conversely, V. volvacea, which prefers high cellulose-, low lignin-containing substrates produces a family of cellulolytic enzymes including at least five endoglucanases, five cellobiohydrolases and two -glucosidases, but none of the recognised lignin-degrading enzymes.J.A. Buswell, Y.J. Cai, S.T. Chang, J.F. Peberdy and S.Y. Fu are with the Department of Biology, The Chinese University of Hong Kong, Shatin, Hong Kong. J.A. Buswell and S.T. Chang are also with the Centre for International Services to Mushroom Biotechnology, The Chinese University of Hong Kong, Shatin, Hong Kong. J.F. Peberdy is also with the Department of Life Science, University of Nottingham, United Kingdom. S.Y. Fu is also with, and H.-s. Yu is with the Institute of Chemistry, Academia Sinica, Guangzhou, China.     

Effect of lignin-derived phenolic monomers on the growth of the edible mushrooms Lentinus edodes,Pleurotus sajor-caju and Volvariella volvacea

Pleurotus sajor-caju was generally more tolerant to lignin-related phenolic monomers and tannin derivatives than Lentinus edodes and the straw mushroom, Volvariella volvacea. Several phenols, at up to 5 mM, enhanced mycelial growth of P. sajor-caju. No clear pattern was evident when the effects of phenols and tannins on the growth of V. volvacea and L. edodes were compared, but the lower concentrations of 4-hydroxybenzaldehyde and vanillin which were tested were markedly more toxic to the straw mushroom. The distribution of phenolic monomers and tannin derivatives in the agricultural wastes used for mushroom cultivation may be an important growth determinant. However, the differences in the growth inhibition profiles of L. edodes, P. sajor-caju and V. volvacea suggest that, alone, the effect of these compounds on fungal growth is unlikely to account for the varying abilities of the three mushroom species to grow and fruit on a particular lignocellulosic substrate.     

Molecular Cloning,Sequencing, and Expression of a <Emphasis Type="SmallCaps">l</Emphasis>-Glutamine <Emphasis Type="SmallCaps">d</Emphasis>-Fructose 6-Phosphate Amidotransferase Gene from <Emphasis Type="Italic">Volvariella volvacea</Emphasis>

Using 3′-RACE and 5′-RACE, we have cloned and sequenced the genomic gene and complete cDNA encoding l-glutamine d-fructose 6-phosphate amidotransferase (GFAT) from the edible straw mushroom, Volvariella volvacea. Gfat contains five introns, and encodes a predicted protein of 697 amino acids that is homologous to other reported GFAT sequences. Southern hybridization indicated that a single gfat gene locus exists in the V. volvacea genome. Recombinant native V. volvacea GFAT enzyme, over-expressed using Escherichia coli and partially purified, had an estimated molecular mass of 306 kDa and consisted of four equal-sized subunits of 77 kD. Reciprocal plots revealed K _m values of 0.55 and 0.75 mM for fructose 6-phosphate and l-glutamine, respectively. V. volvacea GFAT activity was inhibited by the end-product of the hexosamine pathway, UDP-GlcNAc, and by the glutamine analogues N ³-(4-methoxyfumaroyl)-l-2,3-diaminopropanoic acid and 2-amino-2-deoxy-d-glucitol-6-phosphate.     

In vitro evaluation of solvent extracted compounds from edible macromycetes against phytopathogenic fungi

Extraction of antimicrobial compounds from medicinal mushrooms, namely, Auricularia polytricha (Jew's ear), Lentinulla edodes (Shiitake) and Volvariella volvacea (paddy straw), has been done with different solvent systems (chloroform, ethyl acetate, ether and methanol) and tested against a wide range of phytopathogens by filter paper disk assay. All the three basidiomycetes inhibited the phytopathogens tested so far. This enumeration was based on the number of organisms inhibited and the diameter of inhibitory zones produced. From the results obtained, it could be observed that ethyl acetate was the best solvent for extracting antimicrobial substances from these medicinal mushrooms. Thin layer chromatography revealed the presence of several compounds with different Retention Factor (RF) values. Sodium dodecyl sulphate–polyacrylamide gel electrophoresis profiling of V. volvacea showed the presence of several antimicrobial proteins with various molecular weights. Western blotting revealed the presence of thaumatin-like glycoproteins of molecular weight more than 45 kDa.     

A (1→6)-β-glucan from a somatic hybrid of Pleurotus florida and Volvariella volvacea: isolation, characterization, and study of immunoenhancing properties

A water-soluble immunoenhancing polysaccharide was isolated from the aqueous extract of fruit bodies of somatic hybrid (Pflo Vv5 FB), obtained through protoplast fusion between Pleurotus florida and Volvariella volvacea strains. On the basis of acid hydrolysis, the polysaccharide was found to contain glucose only. Methylation analysis, periodate oxidation along with ¹H, DEPT-135, and ¹³C NMR spectroscopy, including two-dimensional TOCSY, DQF-COSY, NOESY, ROESY, 1H,13C-HMQC, and HMBC experiments showed that the polysaccharide was a (1→6)-β-d-glucan, which was not a constituent of any of the parent mushrooms previously reported.This glucan stimulated the macrophages, splenocytes, and thymocytes.     

Nutritional studies on species and mutants of Lepista,Cantharellus, Pleurotus and Volvariella

The purpose of this study is to determine the ability of select Agaricales species to utilize various sources of carbon, nitrogen, vitamins, and growth hormones. Fungi selected for the studies include:Cantharellus clavatus Fries,C. cibarius Fries,Lepista nuda (Bull. ex. Fries)Cooke,Pleurotus ostreatus (Jacq. ex. Fries)Kummer, andVolvariella volvacea (Bull. ex. Fries)Singer. Three strains ofC. cibarius and one mutant ofV. volvacea (V135), V134, were employed to determine if nutritional requirement differences occurred. One species,V. volvacea, is grown commercially as a cottage industry in the Orient (Alicbusan &Ela, 1961) while the other species currently have no commercial value. All species studied possess pleasing flavors and have potential use in the mushroom production industry.A literature compilation of the nutritional regulation of basidiocarp formation and vegetative growth of Agaricales was made with specific mention to the named species (Volz &Beneke, 1969). Recent nutritional studies with one or more of the specific species include those byYusef &Allam (1967), andEger (1970).     

新疆阿魏菇与杏鲍菇中氨基酸含量的分析研究

对新疆阿魏菇与杏鲍菇的氨基酸成分进行系统分析,结果显示新疆本地的阿魏菇和杏鲍菇中17种氨基酸含量丰富,种类齐全,阿魏菇中氨基酸总量为16.80%高于杏鲍菇中氨基酸总量(14.34%);两种菇中必需氨基酸含量(EAA)占总氨基酸含量的比例分别为34.72%和37.92%。新疆阿魏菇与杏鲍菇种富含人体必需的氨基酸,可作为饮食氨基酸来源的重要补充。     

Mycelium growth kinetics and optimal temperature conditions for the cultivation of edible mushroom species on lignocellulosic substrates

The influence of environmental parameters on mycelial linear growth ofPleurotus ostreatus, P. eryngii, P. pulmonarius, Agrocybe aegerita, Lentinula edodes, Volvariella volvacea andAuricularia auricula-judae was determined in two different nutrient media in a wide range of temperature, forming the basis for the assessment of their temperature optimaV. volvacea grew faster at 35°C,P. eryngii at 25°C,P. ostreatus andP. pulmonarius at 30°C,A. aegerita at 25 or 30°C andA. auricula-judae at 20 or 25°C depending on the nutrient medium used andL. edodes at 20 or 30°C depending on the strain examined. The mycelium extension rates were evaluated on seven mushroom cultivation substrates: wheat straw, cotton gin-trash, peanut shells, poplar sawdust, oak sawdust, corn cobs and olive press-cake. The mycelium extension rates (linear growth and colonization rates) were determined by the ‘race-tube’ technique, and were found to be the highest on cotton gin-trash, peanut shells and poplar sawdust forPleurotus spp. andA. aegerita. Wheat straw, peanut shells and particularly cotton gin-trash supported fast growth ofV. volvacea, whereas wheat straw was the most suitable substrate forL. edodes andA. auricula-judae. Supplemented oak sawdust and olive press-cake were poor substrates for most species examined, white almost all strains performed adequately on corn cobs.     

Protein enrichment and digestibility of soft rush (<Emphasis Type="Italic">Juncus effusus</Emphasis>) and rice residues using edible mushrooms <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> and <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis>

Pleurotus species are found to be among the most efficient lignocellulolytic types of white-rot fungi. Rice is the main grain cultivated in the extreme south of Brazil. Defatted rice bran and straw are by-products of low aggregate value. Soft rush (Juncus effusus) is a common native plant also very abundant in the region. In the present work, we evaluated changes in substrate composition after growth of two white-rot fungal species: Pleurotus ostreatus and Pleurotus sajor-caju, aiming to increase protein content and digestibility from substrates through solid fermentations and obtain edible mushrooms of high aggregate value. For that, defatted rice bran, defatted rice straw and soft rush were utilized as substrate. The influence of the variables thermal treatment temperature of substrate, substrate moisture and concentration were evaluated on the protein content, digestibility and biological efficiency. The highest protein enrichment of rice bran in P. sajor-caju-fermented medium was due the fact that there was no fructification in these media, while for the P. ostreatus-fermented medium, part of the synthesized protein was converted into mushrooms. The highest protein enrichments were verified in medium with 80% moisture and 25% soft rush (47.1% using P. ostreatus and 49.0% using P. sajor-caju). A higher digestible protein increase was obtained for both species in media with 70% moisture and 25% soft rush.     

Therapeutic potential of culinary-medicinal mushrooms for the management of neurodegenerative diseases: diversity,metabolite, and mechanism

Mushrooms have long been used not only as food but also for the treatment of various ailments. Although at its infancy, accumulated evidence suggested that culinary-medicinal mushrooms may play an important role in the prevention of many age-associated neurological dysfunctions, including Alzheimer’s and Parkinson’s diseases. Therefore, efforts have been devoted to a search for more mushroom species that may improve memory and cognition functions. Such mushrooms include Hericium erinaceus, Ganoderma lucidum, Sarcodon spp., Antrodia camphorata, Pleurotus giganteus, Lignosus rhinocerotis, Grifola frondosa, and many more. Here, we review over 20 different brain-improving culinary-medicinal mushrooms and at least 80 different bioactive secondary metabolites isolated from them. The mushrooms (either extracts from basidiocarps/mycelia or isolated compounds) reduced beta amyloid-induced neurotoxicity and had anti-acetylcholinesterase, neurite outgrowth stimulation, nerve growth factor (NGF) synthesis, neuroprotective, antioxidant, and anti-(neuro)inflammatory effects. The in vitro and in vivo studies on the molecular mechanisms responsible for the bioactive effects of mushrooms are also discussed. Mushrooms can be considered as useful therapeutic agents in the management and/or treatment of neurodegeneration diseases. However, this review focuses on in vitro evidence and clinical trials with humans are needed.     

Composition and Expression of Genes Encoding Carbohydrate-Active Enzymes in the Straw-Degrading Mushroom Volvariella volvacea

Volvariella volvacea is one of a few commercial cultivated mushrooms mainly using straw as carbon source. In this study, the genome of V. volcacea was sequenced and assembled. A total of 285 genes encoding carbohydrate-active enzymes (CAZymes) in V. volvacea were identified and annotated. Among 15 fungi with sequenced genomes, V. volvacea ranks seventh in the number of genes encoding CAZymes. In addition, the composition of glycoside hydrolases in V. volcacea is dramatically different from other basidiomycetes: it is particularly rich in members of the glycoside hydrolase families GH10 (hemicellulose degradation) and GH43 (hemicellulose and pectin degradation), and the lyase families PL1, PL3 and PL4 (pectin degradation) but lacks families GH5b, GH11, GH26, GH62, GH93, GH115, GH105, GH9, GH53, GH32, GH74 and CE12. Analysis of genome-wide gene expression profiles of 3 strains using 3′-tag digital gene expression (DGE) reveals that 239 CAZyme genes were expressed even in potato destrose broth medium. Our data also showed that the formation of a heterokaryotic strain could dramatically increase the expression of a number of genes which were poorly expressed in its parental homokaryotic strains.     

Identification and expression analysis of a new glycoside hydrolase family 55 exo-β-1,3-glucanase-encoding gene in Volvariella volvacea suggests a role in fruiting body development

The edible straw mushroom Volvariella volvacea is an important crop in South East Asia and is predominantly harvested in the egg stage. Rapid stipe elongation and cap expansion result in a swift transition from the egg to elongation and maturation stage, which are subjected to fast senescence and deterioration. In other mushrooms, β-1,3-glucanases have been associated with degradation (softening) of the cell wall during stipe elongation and senescence. We present a new glycoside hydrolase family 55 (GH55) exo-β-1,3-glucanase gene, exg2, and highly conserved deduced EXG2 protein. The 3D model and presumed catalytic residues of V. volvacea EXG2 are identical to Lentinula edodes EXG2 and Phanerochaete chrysosporium Lam55A, supporting similar enzymatic functions. In addition to previous association to stipe elongation and senescence, our data clearly indicates a role for cap (pileus) expansion. Digital gene expression, quantitative PCR and isobaric tags for relative and absolute quantification analysis showed low exg2 and EXG2 levels in primordia, button, egg and elongation stages and significantly increased levels in the maturation stage. Subsequent relative quantitative PCR analysis designated expression of exg2 to the stipe in the elongation stage and to the pileus and stipe in the maturation stage. EXG2 cell wall softening activity, close correlation of exg2 expression with the principal expanding mushroom tissues and a strong conservation of expression patterns and protein sequences in other mushrooms, make V. volvacea exg2 an important candidate for future studies on mechanisms of fruiting body expansion and senescence causing commodity value loss.     

The composition of volatile components of cepe (<Emphasis Type="Italic">Boletus edulis</Emphasis>) and oyster mushrooms (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis>)

The composition of aroma compounds in cooked and canned cepe (Boletus edulis) and in cooked oyster mushrooms (Pleurotus ostreatus) is studied using capillary gas chromatography and chromatographymass spectrometry. It is found that unsaturated alcohols and ketones containing eight atoms of carbon determine the aroma of raw mushrooms and take part in the formation of the aroma of cooked mushrooms as well. The content of these compounds was the highest in canned cepes. In oyster mushrooms, the concentration of these alcohols and ketones was lower in comparison with cepes. The content of aliphatic and aromatic aldehydes was much higher in oyster mushrooms. Volatile aliphatic and heterocyclic Maillard reaction products and isomeric octenols and octenones formed the aroma of cooked and canned mushrooms.     

Isolation of protoplasts from edible fungi

Summary Mycelium from the periphery of actively growing colonies on cellophane and from shake flask cultures was used for the isolation of protoplasts from strains ofAgaricus bisporus,Auricularia auricula, Lentinus edodes, Pleurotus sajor-caju, Volvariella bombycina andV. volvacea. The mycelial cells were treated with two mycolytic enzymes, Novozym 234 or lywallzyme. Protoplasts were produced from all the edible fungi tested.Pleurotus sajor-caju gave the highest yield (3.84×10⁷/ml), followed byAu. auricula (7.46×10⁶/ml),Ag. bisporus (2.16×10⁶/ml) andV. volvacea (1.92×10⁶/ml), when treated with lywallzyme.Agaricus bisporus gave the smallest yield of protoplasts when Novozym 234 was used. The effects of different molarities of osmotic stabilizers were also studied. The cellophane method is simple and quick and can be used as a screening procedure. The yields of protoplasts obtained from liquid cultures were usually higher than those from cellophane cultures.

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Aislamiento de protoplastos a partir de setas comestibles

Resumen Para el aislamiento de protoplastos de cepas deAgaricus bisporus, Auricularia auricula,Lentinus edodes, Pleurotus sajor-caju, Volvariella bombycina, y V. volvacea se utilizo micelio periferico de colonias que estaban en crecimiento activo en celofan y en frascos de agitación. Las celulas del micelio se tratarón con dos enzymas mycoliticos: Novozym 234 y lywallzyme. Todas las setas comestibles ensayadas produjerón protoplastos. Mediante tratamiento con lywallzyme la mejor cosecha de protoplastos la proporcionoP. sajor-caju (3.84×10⁷/ml) seguido porAu. auricula (7.46×10⁶/ml),Ag. bisporus (2.16×10⁶/ml) yV. volvacea (1.92×10⁶/ml). Con Novozym 234Ag. bisporus fué la de menor rendimiento. También se estudiarón los efectos de distintos estabilizadores osmoticos a diferentes molaridades.

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Isolement de protoplastes à partir de champignons comestibles

Résumé Du mycélium prélevé à la périphérie de colonies dévoloppées sur cellophane ou à partir de cultures liquides agitées a été utilizé pour la préparation de protoplastes d'Agaricusbisporus,Auricularia auricula, Lentinus edodes, Pleurotus sajor-caju, Volvariella bombycina etV. volvacea. Les cellules mycéliales ont été traitées par deux enzymes mycolytiques, Novozym 234 et lywallzyme. Par traitement avec le lywallzyme, des protoplastes ont été obtenus avec tous les champignons comestibles étudiés,P. sajor-caju donnant le rendement le plus élevé (3.84×10⁷/ml), suivi parAu. auricula (7.46×10⁶/ml,Ag. bisporus (2.16×10⁶/ml etV. volvacea 1.92×10⁶/ml). Avec le Novozym 234,Ag. bisporus a donné le rendement en protoplastes le moins élevé. Les effets de différentes molarités de stabilisateurs osmotiques ont également été étudiés. La méthode de la cellophane est simple et rapide et peut être employée comme procédé de degrossissage. Les rendements en protoplastes obtenus à partir des cultures liquides sont habituellement plus élevés que ceux à partir des cultures sur cellophane.

     

Tolerance of tannin by the shiitake mushroom,Lentinus edodes

Summary Tannin at 1% (w/v) did not inhibit the growth ofLentinus edodes, but did inhibitPleuroius florida, P. sajor-caju, P. cystidosus, Agaricus bisporus andVolvariella volvacea. The inhibition was not due to its acidity.

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Tolérance de Lentinus edodes aux ajouts de tannin

Résumé Le tannin à la concentration de 1% (p/v) n'inhibe pas la croissance deLentinus edodes, mais inhibe celle dePleurotus florida, P. sajor-caju, P. cystidosus, Agaricus bisporus, etVolvariella volvacea. L'inhibition n'est pas due à son acidité.

     

First Report about Mineral Content,Fatty Acids Composition and Biological Activities of Four Wild Edible Mushrooms

The goal of this research was a comprehensive analysis of four wild edible mushroom species, Cantharellus cinereus, Clavariadelphus pistillaris, Clitocybe nebularis and Hygrocybe punicea, which have not been analyzed so far. Extracts of different polarities have been prepared and evaluated for their antioxidant activities by DPPH, ABTS, FRAP, TRP and CUPRAC methods. For all extracts, total phenolic content was determined. Based on the analysis, it was shown that solvent type had a significant effect on the antioxidant capacities of mushroom extracts, so water extracts showed the highest activity. Furthermore, the analysis includes determination of mineral composition, fatty acid profiles and antimicrobial activity. Unsaturated fatty acids, which are very important for human health, are dominant in the studied mushroom species. Linoleic and oleic acid consist of over 50 % of the total fatty acid composition. Seventeen biologically important and toxic elements have been analyzed by ICP‐OES and ICP‐MS and results showed that the element concentrations were species‐dependent. Also, it has been found that analyzed mushrooms did not show any antimicrobial activity. Chemometric analysis was used to understand the connection between the extracts of different polarities.     

CaMV 35S promoter directs β-glucuronidase expression in Ganoderma lucidum and Pleurotus citrinopileatus

The cauliflower mosaic virus (CaMV) 35S promoter has been most commonly used in plant transformation studies, but its activity in mushrooms has not been reported. p301-b is a binary vector containing a bialaphos resistance gene driven by the promoter of Lentinus edodes glyceraldehyde-3-phosphate dehydrogenase (GPD) gene. CaMV 35S-GUS was inserted into p301-b, and the resulting construct p301-bG was transformed to protoplasts of Ganoderma lucidum and basidiospores of Pleurotus citrinopileatus. GUS activity was observed in the transformants, indicating that CaMV 35S promoter can direct expression of exogenous gene in the mushrooms. This is the first report on the application of CaMV 35S promoter in genetic modification of mushrooms.