Profile of the extracellular lignocellulolytic enzymes activities as a tool to select the promising strains of Volvariella volvacea (Bull. ex Fr.) sing

Out of 26 strains of Volvariella volvacea used, 18 were of ‘typical’ type and possessed all the characteristics of a normal V. volvacea mycelium, while the rest 4 ‘atypical’ type strains showed completely distinct mycelial growth characteristics. The remaining 4 strains grew very slowly and exhibited growth characteristics of single spore isolates of V. volvacea. Strains varied in their extracellular lignocellulolytic activities and strains; OE-274, OE-272 and OE-210 with high ligninase enzymes (laccase and polyphenol oxidase) activities, gave highest mushroom yield on pasteurized paddy straw substrate. On the composted paddy straw substrate, additional two strains, OE-213 and OE-215 with lower activities of ligninases also gave higher mushroom yield. Mushrooms were harvested 3 to 4 d early from the composted substrate than on the pasteurized substrate. Activities of endoglucanase, laccase and polyphenol oxidase were found to be more crucial for mushroom yield on pasteurized substrate, while xylanase and β-glucosidase were more important for composted substrate. Strains also varied in their fruiting bodies quality and the substrate used for mushroom cultivation also affected the fruiting body quality. The superior yielding strains varied in shape, size, weight, colour and contents of sodium and potassium in their fruiting bodies; while contents of carbon, calcium and protein did not vary much with the strains.     

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.     

Effects of different levels of wheat bran, rice bran and maize powder supplementation with saw dust on the production of shiitake mushroom (Lentinus edodes (Berk.) Singer)

The cultivation of shiitake mushroom (Lentinus edodes) is increasing rapidly in Bangladesh due to its nutritional and medicinal importance with excellent flavor and longer shelf life. With the aim of increased production, we have cultivated L. edodes on saw dust (SD) supplemented with different levels (10%, 15%, 20%, 25%, 30%, 35% and 40%) of wheat bran (WB), rice bran (RB), maize powder (MP) and their combination (WB+RB+MP = 1:1:1) to investigate the growth, yield and quality of this mushroom. Most of the growth, yield and quality parameters varied significantly when mushrooms were cultivated with different levels of supplementation. The yield of mushroom was increased with the level of each supplementation upto a certain level, and then decreased. SD supplemented with 25% WB produced the highest number of fruiting bodies (34.8/500 g packet), highest biological yield (153.3/500 g packet), and biological efficiency (76.6%) of L. edodes. But the yield of the best quality mushroom was observed on SD with 40% WB supplementation; however, the qualities were not always supplementation dose dependent. In this study, we report that 25% WB supplementation with SD may be very effective for higher yield and 40% WB supplementation for better quality of L. edodes.     

Crystallization and preliminary X-ray analysis of volvatoxin A2 from Volvariella volvacea

Volvatoxin A2, an ion channel disturbed cardiotoxic and hemolytic protein from the edible mushroom, Volvarilla volvacea, has been crystallized by the vapor diffusion method using polyethylene glycol 4000 and ammonium sulfate in sodium acetate buffer pH 4.6. The best crystals belong to the monoclinic space group C2 with unit cell dimensions a = 155.25 Å, b = 58.06 Å, c = 116.92 Å, and β = 119.5°. These crystals diffract to at least 2.2 Å and there are four molecules of molecular weight 24 kDa per asymmetric unit with a solvent content of 48%.     

Dietary effect of Pleurotus eryngii on biochemical function and histology in hypercholesterolemic rats

This work was conducted to investigate diet supplement of king oyster mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-week old female Sprague–Dawley albino rats were divided into three groups of 10 rats each. The feeding of 5% powder of the fruiting bodies of Pleurotus eryngii to hypercholesterolemic rats reduced their plasma total cholesterol, triglyceride, low-density lipoprotein, total lipid, phospholipids, and LDL/HDL ratio by 24.05%, 46.33%, 62.50%, 24.63%, 19.22%, and 57.14%, respectively. Mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no adverse effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, and enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that P. eryngii significantly reduced plasma β and pre-β-lipoprotein, while increased α-lipoprotein. A histological study of hepatic cells by conventional hematoxylin–eosin and oil red O staining showed normal findings for mushroom-fed hypercholesterolemic rat. The present study suggests that 5% P. eryngii diet supplement provided health benefits by acting on the atherogenic lipid profile in hypercholesterolaemic rats. Therefore, king oyster mushroom could be recommended as a natural cholesterol lowering substance within the human diet.     

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.     

人工疏蕾对杏鲍菇产量及品质的影响

【背景】工厂化栽培中,杏鲍菇是不定点出菇,通常采用人工疏蕾的方法来实现对子实体数目的控制,但目前关于人工疏蕾保留子实体的数目无具体的研究。【目的】通过人工疏蕾的方式,研究保留不同子实体数目对杏鲍菇产量及品质的影响。【方法】对各处理组进行基质利用情况测定,对采收后各组子实体进行产量、形态指标及质构特性的测定。【结果】保留3—4个子实体的基质利用率较高,在生产实际中对成本的浪费较少;保留不同子实体数目对子实体形态指标、单菇重量及单包产量都有显著影响,保留1个子实体时,单菇重量最大但单包产量最低,保留4个子实体时,单包产量较高且子实体形态一致;从质构特性来看,保留子实体数目为4个时,杏鲍菇子实体品质最好、口感最佳。【结论】在杏鲍菇工厂化生产的人工疏蕾环节,保留子实体数目为3—4个时可以减少对成本的浪费,获得产量较高、品质较好的产品。     

Influence of spatio-temporal resource availability on mushroom mite diversity

Although biodiversity in nature is of fundamental importance because it improves the sustainability of ecosystems, communities of microscopic organisms are generally excluded from conservation targets for biodiversity. Here, I hypothesize that mushroom mite species richness is correlated with both spatial (i.e., mushroom size) and temporal (i.e., longevity of fruiting bodies) resource availability. I collected fruiting bodies in an old-growth forest over 4 years to collect mites and insects inhabiting the mushrooms. Mites were collected from 47 % of the fruiting bodies and approximately 60 % of the mite species were collected only once. Mite species richness was significantly correlated with the availability of long-lasting fruiting bodies. For example, bracket fungi contained more mite species than ephemeral fruiting bodies. Insect presence was also correlated with mushroom mite richness, probably as phoretic hosts and food resources for predacious mites. On the other hand, mushroom size seemed to be less important; small fruiting bodies sometimes harbored several mite species. Although mite species richness was correlated with mushroom species richness, mushroom specificity by mites was not clear except for a preference for long-lasting fruiting bodies. Therefore, I suggest that a constant supply of coarse woody debris is crucial for maintaining preferred resources for mushroom mites (e.g., bracket fungi) and their associated insects (mycophilous and possibly saproxylic insects).     

Comparative studies on the diversity of the edible mushroom Pleurotus eryngii: ITS sequence analysis, RAPD fingerprinting, and physiological characteristics

Verification of Pleurotus eryngii strains was assessed using ITS sequence analysis and RAPD fingerprinting. Sequence analysis of the ITS1–5.8S rDNA–ITS2 region of 24 strains of Pleurotus sp., which consisted of 22 strains of P. eryngii and the control strains P. ostreatus and P. ferulae, demonstrated that the DNA regions share mostly 99 % sequence identity, indicating that sequence-based analysis is not applicable for the verification of closely related mushroom strains. To verify the mushroom strains using RAPD, we amplified DNA fragments from the total cellular DNA of 24 mushroom strains with 18 different random primers, yielding 538 distinct DNA fragments ranging from 200–4000 bp. Analysis of the DNA fragment pattern showed that the 22 P. eryngii strains were clearly distinguished from the control strains P. ostreatus and P. ferulae, and could be categorized into five subgroups. Subsequent physiological studies on the development of fruiting bodies demonstrated the close correlation of the RAPD-based grouping with the phenotypical characteristics of mushroom fruiting bodies.     

Cultivation of different strains of king oyster mushroom (Pleurotus eryngii) on saw dust and rice straw in Bangladesh

Pleurotus eryngii is a popular mushroom due to its excellent consistency of cap and stem, culinary qualities and longer shelf life. In Bangladesh, where Pleurotus mushrooms are very popular, P. eryngii may take position among the consumers, but currently this mushroom is not cultivated in large scale there. In this study, 3 strains of P. eryngii such as Pe-1 (native to Bangladesh), Pe-2 (germplasm collected from China) and Pe-3 (germplasm collected from Japan) were cultivated on saw dust and rice straw and their growth and yield parameters were investigated. Pe-1 on saw dust showed the highest biological yield and efficiency (73.5%) than other strains. Also, the mycelium run rate and number of fruiting bodies were higher in Pe-1 than other two strains. The quality of mushroom strains was near about similar. On saw dust, the yield and efficiency were better than those cultivated on rice straw, however, on straw; the mushroom fruiting bodies were larger in size. This study shows the prospects of P. eryngii cultivation in Bangladesh and suggests further study in controlled environment for higher yield and production.     

Evaluation of <Emphasis Type="Italic">Volvariella volvacea</Emphasis> Strains for Yield and Diseases/Insect-Pests Resistance Using Composted Substrate of Paddy Straw and Cotton Mill Wastes

Out of the 3 parent strains and 4 single spore isolates of Volvariella volvacea evaluated, strain, OE-274 gave earliest yield in 11.25–11.50 days post-spawning in all 4 trials. The yield varied in different strains in different trials and it was highest in strain, OE-272 in trial 1, SSI, OE-55-08 in trial 2, and strain, OE-274 in trial 3 and 4. In overall average, highest yield was in strain, OE-272, closely followed by strain, OE-274. The number of fruiting bodies per q substrate also varied in different strains in different trials. Highest numbers were in strain, OE-272, SSIs, OE-55-08 and OE-12-22, and strain, OE-210 in trial 1, 2, 3 and 4, respectively. Highest fruiting body wt was in strain, OE-274 in all 4 trials. The yield during different weeks of cropping varied in different strains but invariably it was highest in first week, which accounted for 60–70% of the total yield. The fruiting bodies of strain, OE-274 were of bigger size, brownish, toughest and with least tendency of veil opening, while that of strain, OE-272 and SSI, OE-55-08 were whitish to grayish-white, oblong, medium size, delicate and lesser tendency of veil opening. The strain, OE-274 and SSI, OE-55-08 exhibited higher resistance against the growth of competitor moulds and infestations of insect-pests, while strain, OE-272 exhibited highest susceptibility to insect-pests infestation.     

Cultivation and utility of <Emphasis Type="Italic">Piptoporus betulinus</Emphasis> fruiting bodies as a source of anticancer agents

Piptoporus betulinus is a wood-rotting basidiomycete used in medicine and biotechnology. However, to date, no indoor method for cultivation of this mushroom fruiting bodies has been developed. Here we present the first report of successful production of P. betulinus mature fruiting bodies in artificial conditions. Four P. betulinus strains were isolated from natural habitats and their mycelia were inoculated into birch sawdust substrate supplemented with organic additives. All the strains effectively colonized the medium but only one of them produced fruiting bodies. Moisture and organic supplementation of the substrate significantly determined the fruiting process. The biological efficiency of the P. betulinus PB01 strain cultivated on optimal substrate (moisture and organic substance content of 55 and 65 and 25 or 35 %, respectively) ranged from 12 to 16 %. The mature fruiting bodies reached weight in the range from 50 to 120 g. Anticancer properties of water and ethanol extracts isolated from both cultured and nature-derived fruiting bodies of P. betulinus were examined in human colon adenocarcinoma, human lung carcinoma and human breast cancer cell lines. The studies revealed antiproliferative and antimigrative properties of all the investigated extracts. Nevertheless the most pronounced effects demonstrated the ethanol extracts, obtained from fruiting bodies of cultured P. betulinus. Summarizing, our studies proved that P. betulinus can be induced to fruit in indoor artificial culture and the cultured fruiting bodies can be used as a source of potential anticancer agents. In this respect, they are at least as valuable as those sourced from nature.     

New xanthine oxidase inhibitors from the fruiting bodies of Tyromyces fissilis

Excessive uric acid production, which causes gout and hyperuricemia, can be blocked by inhibiting xanthine oxidase (XO). However, some agents to block on XO often cause side effects, thereby necessitating the identification of new inhibitors. During the screening of XO inhibitors from various mushroom extracts, we found that a methanolic extract of the fruiting bodies of Tyromyces fissilis, an inedible and non-toxic fungus, showed inhibitory activity. Both n-hexane and ethyl acetate layers, obtained by partitioning this extract exhibited XO inhibitory activity. Subsequently, using an activity-guided separation method, eight active compounds (1–8) were isolated. The structures of five of the new compounds, 2–4, 6, and 7, were elucidated by spectral analysis and chemical derivatization. All compounds had a salicylic acid moiety with an aliphatic group at the C-6 position. Notably, 2-hydroxy-6-pentadecylbenzoic acid (1) showed the highest level of XO noncompetitive inhibition (58.9 ± 2.2% at 25 µM).     

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.     

Development of antler-type fruiting bodies of Ganoderma lucidum and determination of its biochemical properties

Following the importance of antler-type fruiting bodies of Ganoderma lucidum, in this study, the impact of main growth parameters such as ventilation and light on the development of antler-type fruiting bodies has been investigated together with the determination of physico-chemical properties of antler fruiting bodies. For this, the primordia bags of G. lucidum were kept under controlled ventilation to adjust the CO₂ produced by the mushrooms owing to its respiration under light and dark conditions. The bioactive compounds such as phenolics, flavonoids, water-soluble polysaccharides and ganoderic acid showed a two-fold increase in the antler-type fruiting bodies as compared to normal kidney-shaped fruiting bodies. It is assumed from this study that the antler type fruiting bodies are developed due to restricted ventilation which causes an increase in the level of CO₂ gas in the air as a result of respiration of mushroom. The shape and colour of antler fruiting bodies again dependent on the light provided in the growth chamber. This study also proves that with the manipulation of light and ventilation antler-type fruiting bodies of G. lucidum could be developed with higher quantity of bioactive compounds and with higher antioxidant potential.     

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.     

Mineral content of the fruiting bodies of Pleurotus sajor-caju (FR.) SINGER cultivated on different kinds of biomass

Pleurotus sajor-caju (FR. ) SINGER was cultivated on different organic wastes, namely sericulture waste, Populus deltoides MARSH , and Eupatorium adenophorum SPRENG . Paddy straw was taken as the control and all the data were compared with it. The mineral contents of the fruiting bodies of Pleurotus sajor-caju and the substrates on which the mushroom was grown were analyzed. Among the eight minerals determined (calcium, phosphorus, potassium, magnesium, sodium, iron, manganese and zinc), the potassium content was highest followed by phosphorus, magnesium and sodium. Analysis of the mineral contents of the substrates before cultivation had also been carried out. The mineral contents of the fruiting bodies of Pleurotus sajor-caju were found to be different on different substrates. It was also observed that the mineral contents of the fruiting bodies of Pleurotus sajor-caju increase when cultivated on substrates with higher mineral contents. The maximum mineral contents per 100 g of the substrates before cultivation were Ca – 347 mg; P – 151 mg; K – 1,805 mg; Na – 127 mg; Mg – 227 mg; Fe – 53 mg; Mn – 10 mg and zn – 3.1 mg. The mineral contents of the fruiting bodies of Pleurotus sajor-caju per 100 g ranged as follows: Ca – 25.1 mg to 35.3 mg; P – 448 mg to 602 mg; K – 2,146 mg to 2350 mg; Na – 139 mg to 229 mg; Mg – 153 mg to 224 mg; Fe – 9.74 mg to 20.75 mg; Mn – 2.5 mg to 4.0 mg and Zn – 2.2 mg to 3.1 mg.