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.     

Combining high-throughput sequencing with fruit body surveys reveals contrasting life-history strategies in fungi

Before the recent revolution in molecular biology, field studies on fungal communities were mostly confined to fruit bodies, whereas mycelial interactions were studied in the laboratory. Here we combine high-throughput sequencing with a fruit body inventory to study simultaneously mycelial and fruit body occurrences in a community of fungi inhabiting dead wood of Norway spruce. We studied mycelial occurrence by extracting DNA from wood samples followed by 454-sequencing of the ITS1 and ITS2 regions and an automated procedure for species identification. In total, we detected 198 species as mycelia and 137 species as fruit bodies. The correlation between mycelial and fruit body occurrences was high for the majority of the species, suggesting that high-throughput sequencing can successfully characterize the dominating fungal communities, despite possible biases related to sampling, PCR, sequencing and molecular identification. We used the fruit body and molecular data to test hypothesized links between life history and population dynamic parameters. We show that the species that have on average a high mycelial abundance also have a high fruiting rate and produce large fruit bodies, leading to a positive feedback loop in their population dynamics. Earlier studies have shown that species with specialized resource requirements are rarely seen fruiting, for which reason they are often classified as red-listed. We show with the help of high-throughput sequencing that some of these species are more abundant as mycelium in wood than what could be expected from their occurrence as fruit bodies.     

Correspondence between genet diversity and spatial distribution of above- and below-ground populations of the ectomycorrhizal fungus Hebeloma cylindrosporum

Population studies of ectomycorrhizal fungal species have largely relied upon fruit body (the reproductive organ) sampling. Analysis of the fruit bodies alone supposes that they reflect the present and spatial organization of all below-ground genets (mycorrhizas and extramatrical mycelia). The relation between fruit bodies and ectomycorrhizas was investigated for the basidiomycete agaric Hebeloma cylindrosporum in four Pinus pinaster stands in south-west France. Genet identification was based on the comparison of polymorphisms within a hypervariable segment of the ribosomal intergenic spacer amplified by polymerase chain reaction (PCR) using a H. cylindrosporum species-specific primer. Mycorrhizas were sorted from soil samples collected underneath patches of fruit bodies or patches where fruit bodies had or had not been observed during the years prior to mycorrhiza collection. On average 65% of the 1026 mycorrhizas collected underneath fruit bodies were formed by H. cylindrosporum, whereas only 2% of the 954 collected in places from where fruit bodies were absent were formed by this species. All genotypes identified above ground were also identified below ground. In patches where one genotype formed all or more than 90% of the fruit bodies, the same genotype formed all or a large majority of the mycorrhizas. In patches occupied by several different fruiting genotypes, additional nonfruiting ones could be present on the root systems. In all cases, the mycorrhizas of one genotype were found no more than 10-20 cm away from its corresponding fruit bodies, and fruit body disappearance at a given place was associated with the disappearance of the corresponding mycorrhizas within 1 year. Although there was not a strict coincidence between the total numbers of genets present below ground and of those forming fruit bodies, fruit body analysis for H. cylindrosporum appears to reflect both the genetic diversity and the spatial structure of its below-ground populations. The results obtained also illustrate the rapid turnover of ectomycorrhizal fungal species on the root systems in the absence of any obvious major disturbance of the ecosystem.     

Nutritional induction and suppression of fruiting in Myxococcus xanthus FBa

A defined agar medium (A agar) containing 15 amino acids in concentrations between 0.5 and 2 mm was developed for studying the fruiting cycle of Myxococcus xanthus FBa. Cells grew only vegetatively in this medium unless the initial concentration of one of nine required or stimulatory amino acids was lowered about 50-fold. In the latter circumstance, fruiting bodies developed after several days of vegetative growth. The conclusion was that fruiting occurred when any amino acid required for normal growth became limiting in the environment. High concentrations (10 mm) of phenylalanine, tryptophan, or methionine prevented fruiting without affecting growth. Mutants requiring arginine, thymidine, or adenine could not be induced to fruit by limiting their unique requirement although they responded to the same deprivations which brought about fruiting of the wild type. A histidine auxotroph formed fruiting bodies when histidine was lowered to growth-limiting concentrations, provided that the medium was supplemented with purines. A uracil auxotroph was isolated that, perhaps secondarily, had lost some of the mechanisms which control the formation of fruiting bodies; if uracil was present, it formed fruits even when no amino acid was limiting. No concentration of uracil was sufficient to prevent fruiting. Fruiting bodies were formed when mixtures of the uracil auxotroph and wild-type cells were inoculated on A agar plus uracil, even when 75% of the cells were wild type. Microcysts of both strains were present in the fruiting bodies.     

Spatial and temporal patterns of morel fruiting

The biotic and abiotic factors conditioning morel fruit body production are incompletely known. We examined spatial and temporal patterns of Morchella esculenta fruiting over five years in a wooded site in Missouri, USA. Fruiting onset was inversely correlated with spring air and soil temperatures, whereas abundance was positively correlated with rain events (>10 mm) during the 30 d preceding fruiting. The two years with the greatest fruiting had the shortest fruiting seasons (6–7 d). Fruiting season length was positively correlated with soil warming, suggesting that a narrow range of optimum soil temperatures favour the explosive production of fruit bodies. All woody stems of at least 1 cm diam were mapped and stem diameter and crown condition were noted. Morel fruit bodies were significantly closer to stems of Carya spp., Tilia americana and Ulmus americana than predicted by the frequencies of these woody species or their contribution to the total basal area on the site. Although intra-annual clustering of fruit bodies was often observed, inter-annual clustering was not. The spatial pattern of M. esculenta fruiting appears to be associated with vegetation pattern, whereas the onset and abundance of fruiting are determined by the interaction of spring temperatures with availability of supporting precipitation.     

不同基质栽培的猴头菌子实体多糖理化性质和体外免疫活性

【背景】猴头菌多糖是猴头菌(Hericium erinaceus)中主要的活性成分,具有提高免疫力、抗肿瘤、降血糖、抗突变、抗衰老等多种功效。目前对于猴头菌多糖的研究主要集中在结构分析上,有关不同基质栽培对于猴头菌子实体多糖理化性质的影响鲜见报道。【目的】研究不同基质栽培的猴头菌子实体多糖的理化性质和体外免疫活性,筛选出高产优质猴头菌的栽培配方。【方法】采用7种不同配方的基质经工厂化栽培获得不同的猴头菌子实体,对其子实体产量和粗多糖含量进行统计和分析测定,并对7种不同基质栽培获得的猴头菌子实体多糖的分子量分布及单糖组成等理化特征进行分析,比较其对刺激RAW264.7巨噬细胞株释放NO的差异。【结果】综合各项指标表明,配方2 (木屑30%,玉米芯40%,棉籽壳15%,玉米粉2%,麸皮6%,米糠5%,石膏1%,石灰1%)和配方7 (玉米芯39%,棉籽壳10%,麸皮10%,米糠30%,大豆皮5%,甜菜渣4%,碳酸钙2%)栽培获得的猴头菌子实体具有较高的多糖含量和活性。【结论】所选配方适合作为栽培猴头菌深加工原料的培养基质。     

Isolation and characterization of developmental variants in fruiting using a homokaryotic fruiting strain ofCoprinus cinereus

Developmental variants in fruiting ofCoprinus cinereus were induced by mutagenizing oidia of the homokaryotic fruiting strain CopD5-12 with UV light. Through screening of 2,696 isolates, 1,018 strains exhibited defects in fruiting and were classified into 8 groups: (1) knotless variants, which fail to form hyphal knots, the first visible sign of fruiting; (2) primordiumless variants, which form hyphal knots but fail to develop fruit-body primordia; (3) maturationless variants, which form fruit-body primordia but do not form mature fruit bodies; (4) elongationless variants, which form mature fruit bodies with short stipes; (5) expansionless variants, which form mature fruit bodies with unexpanded pilei; (6) sporeless variants, which fail to produce black basidiospores, resulting in fruit bodies with white pilei after maturation; (7) compound type, which includes variants exhibiting several of the phenotypes described above; (8) others, including variants that produce a “dark stipe” even under in light/dark conditions, which is formed under continuous darkness in the wild-type. Two elongationless variants were characterized histologically.     

生长年限对段木栽培鲍姆桑黄子实体营养、活性成分及抗氧化活性的影响

本研究以鲍姆桑黄Sanghuangporus baumii为研究对象,通过测定段木栽培一年生、二年生、三年生子实体粗蛋白质、粗脂肪、粗纤维、氨基酸、多糖、三萜、总黄酮、总酚含量和抗氧化活性(ABTS和FRAP),探究生长年限对段木栽培桑黄子实体营养、活性成分及抗氧化活性的影响.结果表明,生长年限对上述10个指标均产生了...     

A common genetic control of dikaryotic and monokaryotic fruiting in the basidiomycete Agrocybe aegerita

Summary In the edible mushroom Agrocybe aegerita (Agaricales) fruit bodies may be formed in both the sexual and asexual cycle. The major difference between the two types of fruit bodies is that the latter are smaller and contain only two spores on each basidium. Sexual fruiting requires the establishment of a dikaryon which is under the control of the well known incompatibility factors A and B. Asexual fruiting starts directly from a monokaryon. In both dikaryotic and monokaryotic fruiting the same two genes (fi ⁺ , fb ⁺ )are responsible for the initiation and differentiation of fruit bodies respectively. This shows that the morphogenetic procedures leading to fruit body formation in higher basidiomycetes are not necessarily correlated with the sexual cycle. These findings are significant for basic and applied research.     

Interactions of heterologous mycelia colonized in the substrate govern fruit body production in the cultivated homobasidiomycete Pholiota nameko

The spawn of cultivated mushrooms are generally produced, propagated, and distributed to growers as a mycelial culture without genetic purification, in which phenotypic variants frequently occur. We investigated how heterologous mycelia present in a spawn influence fruit body production in the cultivated basidiomycete Pholiota nameko. The 'di-mon' dual cultivation of protoplast clones produced mosaic fruit bodies, which could result from the 'di-mon' mating. In the 'di-di' dual cultivation of heterologous strains with different fruiting times, authentic fruit bodies of each dikaryon and chimera showing a feature combining characteristics of the two dikaryons emerged simultaneously. Mycelia isolated from the chimera produced all three types of fruit bodies, indicating unlikeliness of the occurrence of anastomosis. These results suggest that mycelia colonized in the substrate interact with each other and coordinately promote fruit body production in P. nameko. This phenomenon masks a clonal variability that may be surfaced through multiplication and distribution of the spawn, occasionally bringing about abnormal fruiting.     

Fine‐scale spatiotemporal dynamics of fungal fruiting: prevalence,amplitude, range and continuity

Despite the critical importance of fungi as symbionts with plants, resources for animals, and drivers of ecosystem function, the spatiotemporal distributions of fungi remain poorly understood. The belowground life cycle of fungi makes it difficult to assess spatial patterns and dynamic processes even with recent molecular techniques. Here we offer an explicit spatiotemporal Bayesian inference of the drivers behind spatial distributions from investigation of a Swiss inventory of fungal fruit bodies. The unique inventory includes three temperate forest sites in which a total of 73 952 fungal fruit bodies were recorded systematically in a spatially explicit design between 1992 and 2006. Our motivation is to understand how broad‐scale climate factors may influence spatiotemporal dynamics of fungal fruiting within forests, and if any such effects vary between two functional groups, ectomycorrhizal (ECM) and saprotrophic fungi. For both groups we asked: 1) how consistent are the locations of fruiting patches, the sizes of patches, the quantities of fruit bodies, and of prevalence (occupancy)? 2) Do the annual spatial characteristics of fungal fruiting change systematically over time? 3) Are spatial characteristics of fungal fruiting driven by climatic variation? We found high inter‐annual continuity in fruiting for both functional groups. The saprotrophic species were characterised by small patches with variable fruit body counts. In contrast, ECM species were present in larger, but more distinctly delimited patches. The spatial characteristics of the fungal community were only indirectly influenced by climate. However, climate variability influenced overall yields and prevalence, which again links to spatial structure of fruit bodies. Both yield and prevalence were correlated with the amplitudes of occurrence and of fruit body counts, but only prevalence influenced the spatial range. Summarizing, climatic variability affects forest‐stand fungal distributions via its influence on yield (amount) and prevalence (occupancy), whereas fungal life‐history strategies dictate fine‐scale spatial characteristics.     

Fruiting of Agaricus bisporus

Several enzymes were assayed in extracts from mycelium-colonised compost during growth and fruiting of Agaricus bisporus (Lange) Imbach. Comparison of changes of enzyme levels in axenic and nonaxenic cultures and in cultures of non-fruiting strains indicated that they were associated directly with the fungal mycelium. Large changes were found in the amounts of laccase and cellulase which were correlated with fruit body development. Laccase concentration increased during mycelial growth and then declined rapidly at the start of fruiting. Cellulase activity could be detected throughout growth but increased at fruiting. No such changes were observed in xylanase, alkaline protease, laminarinase and acid and alkaline phosphatases. Activities of laccase and cellulase were measured in axenic cultures arrested at various stages of fruiting development. Such cultures showed that the changes in concentration of laccase and cellulase were associated with the enlargement of fruit bodies.     

Use of coffee grounds for production of Pleurotus ostreatus (Jacq.:Fr.) Kummer

Studies were carried out to screen the industrial strain HK35 of Pleurotus ostreatus for its ability to develop fruiting bodies in solid state cultivation using several substrates containing 17.8 to 55% coffee grounds. Our results showed that only 55% of coffee grounds was used in the substrate without detecting changes in fruiting body or on its biological efficiency of production. The chemical analysis of the caffeine in the substrate showed that this compound decreased about 59% of the mycelium activity, and no caffeine was found in fruiting bodies indicating its degradation by the fungal strain tested.     

The effect of epsilon-aminocaproic acid on biochemical changes in the development of the cellular slime mould Dictyostelium discoideum.

epsilon-Aminocaproic acid (EACA) inhibited the development of Dictyostelium discoideum strain AX2 after the aggregation stage. Biochemical changes that occurred early in development (loss of cellular protein, RNA and carbohydrate; increase in the specific activity of N-acetylglucosaminidase, alpha-mannosidase, threonine deaminase and leucine aminopeptidase) were not affected by concentrations of EACA which blocked development; but biochemical changes that occurred later (synthesis of carbohydrate, increase in the specific activity of UDP-glucose pyrophosphorylase) were inhibited. Spores from fruiting bodies formed in the presence of low concentrations of EACA were larger, more spherical and less able to survive heat treatment than spores from fruiting bodies of control (no EACA) cells.     

Comparison of developmentally controlled glucoamylase from normal and mutant strains of the basidiomycete Schizophyllum commune

During the formation of fruit bodies, the basidiomycete Schizophyllum commune produces glucoamylase activity. DEAE-cellulose chromatography resolves the enzymic activity into a major peak found early in development and a minor peak which appears when the fruit bodies are mature. A mutant homokaryon has been isolated which constitutively produces glucoamylase activity without any fruiting. When purified, the mutant enzyme and the major fruit body enzyme appear to be identical by several physical and kinetic parameters including molecular weight, temperature sensitivity, and K_m.Supported in part by NIH Research Grant AI 09779-03 from the National Institute of Allergy and Infectious Diseases.     

FibA and PilA act cooperatively during fruiting body formation of Myxococcus xanthus

The extracellular matrix (ECM) of Myxococcus xanthus is essential for social (S-) motility and fruiting body formation. An ECM-bound protein, FibA, is homologous to M4 zinc metalloproteases and is important for stimulation by a phosphatidylethanolamine (PE) chemoattractant and for formation of discrete aggregation foci. In this work, we demonstrate that a correlation exists between a reduced ability to respond to PE and the observed defects in fruiting body morphogenesis. Furthermore, the fibA aggregation defect is accentuated by the absence of either PilA, the structural subunit of type IV pili, or DifD, a chemosensory response regulator. The inability to form fruiting bodies is not due to a loss of S-motility, but rather the loss of PilA and pili as pilT fibA mutants form fruiting bodies. The FibA active site residue E342 is important for fruiting body morphogenesis in the absence of PilA. Mutants exhibiting defects in fruiting body morphogenesis also produce fewer viable spores. It is proposed that FibA and PilA act as extracellular sensors for developmental signals.     

Leaf Carbohydrate Status and Enzymes of Translocate Synthesis in Fruiting and Vegetative Plants of Cucumis sativus L

Carbon partitioning in the leaves of Cucumis sativus L., a stachyose translocating plant, was influenced by the presence or absence of a single growing fruit on the plant. Fruit growth was very rapid with rates of fresh weight gain as high as 3.3 grams per hour. Fruit growth was highly competitive with vegetative growth as indicated by lower fresh weights of leaf blades, petioles, stem internodes and root systems on plants bearing a single growing fruit compared to plants not bearing a fruit. Carbon exchange rates, starch accumulation rates and carbon export rates were higher in leaves of plants bearing a fruit. Dry weight loss from leaves was higher at night from fruiting plants, and morning starch levels were consistently lower in leaves of fruiting than in leaves of vegetative plants indicating rapid starch mobilization at night from the leaves of fruiting plants. Galactinol, the galactosyl donor for stachyose biosynthesis, was present in the leaves of fruit-bearing plants at consistently lower concentration than in leaves of vegetative plants. Galactinol synthase, and sucrose phosphate synthase activities were not different on a per gram fresh weight basis in leaves from the two plant types; however, stachyose synthase activity was twice as high in leaves from fruiting plants. Thus, the lower galactinol pools may be associated with an activation of the terminal step in stachyose biosynthesis in leaves in response to the high sink demand of a growing cucumber fruit.     

壳聚糖和ε-聚赖氨酸处理对双孢蘑菇贮藏过程中品质的影响

以延长双孢蘑菇货架期为目标,探讨壳聚糖和ε-聚赖氨酸处理对采后双孢蘑菇在4 ℃贮藏过程中生理特性、营养品质和贮藏特性的影响。结果表明：与对照组双孢蘑菇相比,壳聚糖与ε-聚赖氨酸6:4复配溶液处理能够有效抑制双孢蘑菇表面微生物的生长和多酚氧化酶活性的增加,保持双孢蘑菇子实体较高的L*值和硬度,延缓双孢蘑菇的质量损失和细胞膜透性的升高,减少双孢蘑菇子实体的腐烂,保持较高商品率。在4 ℃条件下,壳聚糖与ε-聚赖氨酸6:4复配溶液对双孢蘑菇的保鲜性能最优,能够有效保持双孢蘑菇的商品品质和延长其贮藏时间。     

A mast fruiting episode of the tropical tree Peltogyne purpurea (Caesalpinaceae) in the Osa Peninsula, Costa Rica

The existence of mast fruiting has not been well documented in the Neotropics. The occurrence of a mast fruiting episode in the population of the tree Peltogyne purpurea in the Osa Peninsula of Costa Rica is described. In February and March of 2000 most of the trees of this species produced a large fruit crop, compared with 1995-1999, when the number of fruit producing trees was very low or zero and those that did bear fruit, did so at a low intensity. In contrast, the fruit crop of 2000 was massive, all trees examined produced fruits and the intensity of fruiting was maximal. There is not enough information on the event for a hypothesis to be formed because the climatic or biological cues that triggered this sporadic flowering are unknown and there is no meteorological data available for this area. Populations with this mode of reproduction may experience local extinction bacause of logging operations.     

Inter‐annual variability of fruit timing and quantity at Nouragues (French Guiana): insights from hierarchical Bayesian analyses

The timing and quantity of fruit production are major determinants of the functioning of a forest community, but simultaneous analyses of both are rare. We analyzed a ten‐year dataset (2001–2011) of fruit production for 45 tree and liana species from the Nouragues rain forest, French Guiana. We developed a hierarchical Bayesian approach to determine variation in the timing and quantity of fruit production. Our analysis accommodates missing censuses and quantifies variation at seasonal and inter‐annual scales. The fruiting peak of 22 of 45 species occurred during the peak of the rainy season, which is typical for central and eastern Amazon. The timing and quantity of fruit production varied substantially across years in most species, with greater variation in quantity than in timing. The timing of fruit production varied from continuously fruiting species to mast fruiting species that had two or more consecutive years without fruit production. Fully 40% of species were mast fruiting species. The seasonal timing and inter‐annual variation in fruiting were unrelated to seed dispersal mode across species. We saw no evidence for directional change in the level of fruit production, the timing of fruit production, or their variances; however, 10 yr is a short record for such analyses.