Selected Species of the Genus Phellinus – Chemical Composition,Biological Activity,and Medicinal Applications

This study presents the current knowledge on chemical composition, biological activity, and possible medicinal applications of Phellinus igniarius, Phellinus pini, Phellinus pomaceus, and Phellinus robustus. These inedible arboreal species are phytopathogens that cause the enzymatic decomposition of wood. These species belong to the medicinal mushrooms and have been known for centuries in the traditional medicine of the Far East. They have been used as an effective remedy for stomach and intestinal ailments, diarrhea, and hemorrhages. Mycochemical studies have proved the presence of polysaccharides, phenolic compounds, and terpenoids. These compounds show biological activities such as anticancer, antioxidant, antiangiogenic, and antiviral. Research studies conducted using modern analytical methods have advanced the knowledge on the potential therapeutic use of compounds isolated not only from the fruiting bodies but also from biomass obtained with in vitro biotechnological methods.     

Quantitative determination of lanostane triterpenes in Fomes officinalis and their fragmentation study by HPLC-ESI

Introduction – The fruit bodies of Fomes officinalis are used for the treatment of coughs, gastric cancer, rheumatism and hydropsia; however, no method is currently available to assess the quality of this medicinal fungus based on quantitative profile of its main triterpenes. Objective – To develop a simple and accurate HPLC‐UV method for the simultaneous quantification of five lanostane‐type triterpenes in the fruit bodies of F. officinalis. Method – Separations were performed on an Agilent Zorbax Eclipse XDB‐C₁₈ column by gradient elution using acetonitrile : formic acid. Analytes were identified by HPLC coupled with electrospray ionisation mass spectrometry experiments. The quantitative HPLC‐UV method was validated for linearity, precision, accuracy and limits of detection and quantification. Results – Calibration curves presented good linear regression (r > 0.9996) within test ranges. The relative standard deviation of this method was less than 1.7% for intra‐ and inter‐day assays and overall recoveries were 96.4–104.1% for the five compounds analysed. The method was successfully applied to the quantification of five triterpenes in 16 samples of F. officinalis collected from different regions. Conclusion – The developed assay could be considered as a suitable quality control method for F. officinalis. Copyright © 2010 John Wiley & Sons, Ltd.     

Production of bioactive metabolites by submerged fermentation of the medicinal mushroom Antrodia cinnamomea: recent advances and future development

Edible and medicinal mushrooms have usually been considered as a sustainable source of unique bioactive metabolites, which are valued as promising provisions for human health. Antrodia cinnamomea is a unique edible and medicinal fungus widespread in Taiwan, which has attracted much attention in recent years for its high value in both scientific research and commercial applications owing to its potent therapeutic effects, especially for its hepatic protection and anticancer activity. Due to the scarcity of the fruiting bodies, the cultivation of A. cinnamomea by submerged fermentation appears to be a promising substitute which possesses some unique advantages, such as short culture time period and its high feasibility for scale-up production. However, the amount of fungal bioactive metabolites derived from the cultured mycelia of A. cinnamomea grown by submerged fermentation is much less than those obtained from the wild fruiting bodies. Hence, there is an urgent need to bridge such a discrepancy on bioactive metabolites between the wild fruiting bodies and the cultured mycelia. The objective of this article is to review recent advances and the future development of the mycelial submerged fermentation of A. cinnamomea in terms of enhancement for the production of fungal bioactive components by the optimization of culture conditions and the regulation of fungal metabolism. This review provides valuable information for further biotechnological applications of A. cinnamomea as well as other mushrooms being the source of bioactive ingredients by submerged fermentation.     

Abundance and viability of fungal spores along a forestry gradient – responses to habitat loss and isolation?

Regional variation in spore deposition and viability was studied for two fungi, Fomitopsis rosea (Alb. & Schwein.: Fr.) P. Karst. and Phlebia centrifuga P. Karst., both confined to old‐growth spruce forests in the boreal zone. Seven regions in Sweden were studied along a north‐south transect in which the historical impact from forestry increases and the amount old forests decreases towards the south. The two southernmost regions were located outside the distribution border of the species. Spore deposition was measured species specifically as heterokaryotisation of homokaryotic mycelia growing on wood discs. There was a significant decline in spore deposition towards the south for both species. F. rosea deposited an average amount of 111 spores m^?2 24 h^?1 in the northernmost region compared to less than 1 spore in the four southernmost regions. The corresponding values for P. centrifuga were 27 spores m^?2 24 h^?1 in the north compared to less than 2 spores in the 4 southernmost regions. No deposition was found south of the distribution borders. The viability of spores from local populations within each region was measured as germination success on nutrient media. Individual fruiting bodies from large populations in the north generally produced spores with higher germinability than fruiting bodies from geographically isolated populations in the central and southern regions. However, there was a high variation among the southern populations. Our data suggest that some populations in mid‐ and south Sweden may suffer from negative genetic effects, possibly associated with fragmentation and loss of habitat. Thus, the combination of low spore deposition and low germinability of spores may be a threat to the long‐term persistence of F. rosea and P. centrifuga in southern Sweden. Several other species may experience the same situation, especially when considering the severe decline of dead wood in Swedish forests.     

Bacteria associated with truffle‐fruiting bodies contribute to truffle aroma

Truffles, symbiotic fungi renown for the captivating aroma of their fruiting bodies, are colonized by a complex bacterial community of unknown function. We characterized the bacterial community of the white truffle Tuber borchii and tested the involvement of its microbiome in the production of sulphur‐containing volatiles. We found that sulphur‐containing volatiles such as thiophene derivatives, characteristic of T. borchii fruiting bodies, resulted from the biotransformation of non‐volatile precursor(s) into volatile compounds by bacteria. The bacterial community of T. borchii was dominated by α‐ and β‐Proteobacteria. Interestingly, all bacteria phyla/classes tested in this study were able to produce thiophene volatiles from T. borchii fruiting body extract, irrespective of their isolation source (truffle or other sources). This indicates that the ability to produce thiophene volatiles might be widespread among bacteria and possibly linked to primary metabolism. Treatment of fruiting bodies with antibacterial agents fully suppressed the production of thiophene volatiles while fungicides had no inhibitory effect. This suggests that during the sexual stage of truffles, thiophene volatiles are exclusively synthesized by bacteria and not by the truffle. At this stage, the origin of thiophenes precursor in T. borchii remains elusive and the involvement of yeasts or other bacteria cannot be excluded.     

<Emphasis Type="Italic">Fomitopsis betulina (</Emphasis>formerly <Emphasis Type="Italic">Piptoporus betulinus)</Emphasis>: the Iceman’s polypore fungus with modern biotechnological potential

Higher Basidiomycota have been used in natural medicine throughout the world for centuries. One of such fungi is Fomitopsis betulina (formerly Piptoporus betulinus), which causes brown rot of birch wood. Annual white to brownish fruiting bodies of the species can be found on trees in the northern hemisphere but F. betulina can also be cultured as a mycelium and fruiting body. The fungus has a long tradition of being applied in folk medicine as an antimicrobial, anticancer, and anti-inflammatory agent. Probably due to the curative properties, pieces of its fruiting body were carried by Ötzi the Iceman. Modern research confirms the health-promoting benefits of F. betulina. Pharmacological studies have provided evidence supporting the antibacterial, anti-parasitic, antiviral, anti-inflammatory, anticancer, neuroprotective, and immunomodulating activities of F. betulina preparations. Biologically active compounds such as triterpenoids have been isolated. The mushroom is also a reservoir of valuable enzymes and other substances such as cell wall (1→3)-α-d-glucan which can be used for induction of microbial enzymes degrading cariogenic dental biofilm. In conclusion, F. betulina can be considered as a promising source for the development of new products for healthcare and other biotechnological uses.     

Tintinnadiol,a sphaeroane diterpene from fruiting bodies of Mycena tintinnabulum

Antifungal metabolites were isolated from fruiting bodies of Mycena tintinnabulum and from mycelia grown in either complex media or on oak wood. Strobilurins were produced in fruiting bodies and in mycelial cultures, whereas a new diterpene, named tintinnadiol, was isolated only from the fruiting bodies. The structure of the new compound was determined by spectroscopic methods. It exhibited cytotoxic effects.     

Spatial Ecology of the Fungal Genus Xylaria in a Tropical Cloud Forest

Fungal symbioses with plants are ubiquitous, ancient, and vital to both ecosystem function and plant health. However, benefits to fungal symbionts are not well explored, especially in non‐mycorrhizal fungi. The Foraging Ascomycete hypothesis proposes that some wood‐decomposing fungi may shift life‐history strategies to endophytism to bridge gaps in time and space between suitable substrates. To test this hypothesis we examine spatial relationships of Xylaria endophytic fungi in the forest canopy with Xylaria decomposer fungi on the forest floor. We sampled for fungi of the genus Xylaria using a spatially explicit sampling scheme in a remote Ecuadorian cloud forest, and concurrently carried out an extensive culture‐based sampling of fungal foliar endophytes. We found 36 species of Xylaria in our 0.5 ha plot, 31 of which were found to only occur as fruiting bodies. All five species of Xylaria found as endophytes were also found as fruiting bodies. We also tested the relationships of both stages of these fungi to environmental variables. Decomposer fungi were differentiated by species‐specific habitat preferences, with three species being found closer to water than expected by chance. In contrast, endophytes displayed no sensitivity to environmental conditions, such as host, moisture, or canopy cover. We found evidence of spatial linkage between life stages in two species. We also demonstrate that direct transmission of endophytes from leaves to woody substrates is possible. These results indicate that endophytism may represent one way for decomposer fungi to escape moisture limitation, and that endophytic fungi may act as sources of dispersal for decomposer fungi consistent with predictions of the Foraging Ascomycete hypothesis.     

Phenolic composition and antioxidant properties of some traditionally used medicinal plants affected by the extraction time and hydrolysis

Introduction – Polyphenolic phytochemicals in traditionally used medicinal plants act as powerful antioxidants, which aroused an increasing interest in their application in functional food development. Objective – The effect of extraction time (5 and 15 min) and hydrolysis on the qualitative and quantitative content of phenolic compounds and antioxidant capacity of six traditionally used medicinal plants (Melissa officinalis L., Thymus serpyllum L., Lavandula officinalis Miller, Rubus fruticosus L., Urtica dioica L., and Olea europea L.) were investigated. Methodology – The content of total phenols, flavonoids, flavan‐3‐ols and tannins was determined using UV/Vis spectrophotometric methods, while individual phenolic acids, flavones and flavonols were separated and detected using HPLC analysis. Also, to obtain relevant data on the antioxidant capacity, two different assays, (2,2‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulphonic acid) (ABTS) radical scavenging and ferric reducing/antioxidant power (FRAP) assays were used. Results – The extraction efficiency of phenolics, as well as the antioxidant capacity of plant extracts, was affected by both prolonged extraction and hydrolysis. The overall highest content of phenolic compounds was determined in hydrolyzed extract of blackberry leaves (2160 mg GAE/L), followed by the non‐hydrolyzed extract of lemon balm obtained after 15 min of extraction (929.33 mg GAE/L). The above extracts also exhibited the highest antioxidant capacity, while extracts of olive leaves were characterized with the lowest content of phenolic compounds, as well as the lowest antioxidant capacity. The highest content of rosmarinic acid, as the most abundant phenolic compound, was determined in non‐hydrolyzed extract of lemon balm, obtained after 15 min of extraction. Although the hydrolysis provided the highest content of polyphenolic compounds, longer extraction time (15 min) was more efficient to extract these bioactives than shorter extraction duration (5 min). Conclusion – The distribution of detected phenolic compounds showed a wide variability with regard to their botanical origin. Examined medicinal plants showed to be a valuable supplement to a daily intake of bioactive compounds. Copyright © 2010 John Wiley & Sons, Ltd.     

Bioluminescence characteristics of the fruiting body of Mycena chlorophos

Bioluminescent fungi are widely distributed on land and most belong to the class Basidomycetes. Light of about 530 nm wavelength maximum is emitted continuously. The molecular basis for the light‐emitting process remains unclear. We investigated the characteristics of the bioluminescence using cultivated fruiting bodies of M. chlorophos. Only fresh fruiting bodies exhibited long‐lasting light emission; rapid decay of light emission was observed with frozen and freeze‐dried samples. Freeze‐dried samples can be stored at room temperature under dry conditions and may be useful for the isolation of luciferin. The light emission of the fresh fruiting bodies was maintained in various buffers at varying pH; it could be stopped with pH 4 acetate buffer and could be recovered at pH 6. The isolation of luciferin from the fresh fruiting bodies might be possible by the control of buffer pH. The effect of temperature on the light emission of fruiting bodies indicated that bioluminescence in M. chlorophos may involve enzymatic reaction(s). The solubilization of bioluminescent components from the fruiting bodies could not be achieved with various surfactants. Copyright © 2011 John Wiley & Sons, Ltd.     

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.     

Ecology of coarse wood decomposition by the saprotrophic fungus <Emphasis Type="Italic">Fomes fomentarius</Emphasis>

Saprotrophic wood-inhabiting basidiomycetes are the most important decomposers of lignin and cellulose in dead wood and as such they attracted considerable attention. The aims of this work were to quantify the activity and spatial distribution of extracellular enzymes in coarse wood colonised by the white-rot basidiomycete Fomes fomentarius and in adjacent fruitbodies of the fungus and to analyse the diversity of the fungal and bacterial community in a fungus-colonised wood and its potential effect on enzyme production by F. fomentarius. Fungus-colonised wood and fruitbodies were collected in low management intensity forests in the Czech Republic. There were significant differences in enzyme production by F. fomentarius between Betula pendula and Fagus sylvatica wood, the activity of cellulose and xylan-degrading enzymes was significantly higher in beech wood than in birch wood. Spatial analysis of a sample B. pendula log segment proved that F. fomentarius was the single fungal representative found in the log. There was a high level of spatial variability in the amount of fungal biomass detected, but no effects on enzyme activities were observed. Samples from the fruiting body showed high β-glucosidase and chitinase activities compared to wood samples. Significantly higher levels of xylanase and cellobiohydrolase were found in samples located near the fruitbody (proximal), and higher laccase and Mn-peroxidase activities were found in the distal ones. The microbial community in wood was dominated by the fungus (fungal to bacterial DNA ratio of 62-111). Bacterial abundance composition was lower in proximal than distal parts of wood by a factor of 24. These results show a significant level of spatial heterogeneity in coarse wood. One of the explanations may be the successive colonization of wood by the fungus: due to differential enzyme production, the rates of biodegradation of coarse wood are also spatially inhomogeneous.     

Triterpenes and Meroterpenes with Neuroprotective Effects from Ganoderma leucocontextum

Ganoderma leucocontextum is a well‐known medicinal mushroom cultivated in the Tibet Plateau of China. Chemistry investigation on the fruiting bodies of this mushroom resulted in the isolation of sixteen secondary metabolites including three new lanostane triterpenes, ganoleucoins Q – S ( 1 – 3 ), as well as thirteen known compounds ( 4 – 16 ). The structures of compounds 1 – 3 were determined by NMR, MS, CD spectral analysis, and chemical derivation method. The neuroprotective effects of compounds 1 – 16 were tested on PC12 cells. Compounds 1 and 2 showed protective effects against the H₂O₂ induced damage with the survival rate of 83.19 ± 0.92%, 73.37 ± 1.25% at the concentration of 200 μm , respectively. Meanwhile, compounds 1 and 2 induced neurite outgrowth at 50 – 200 μm . The results from this study suggested that G. leucocontextum and its metabolites may be potential functional food ingredients for the prevention of neurodegenerative diseases.     

A Comprehensive Characterisation of Rosemary tea Obtained from Rosmarinus officinalis L. Collected in a sub‐Humid Area of Tunisia

Introduction

Rosemary (Rosmarinus officinalis L.) is an aromatic plant common in Tunisia and it is widely consumed as a tea in traditional cuisine and in folk medicine to treat various illnesses. Currently, most research efforts have been focused on rosemary essential oil, alcoholic and aqueous extracts, however, little is reported on rosemary infusion composition.

Objective

To investigate compounds present in rosemary tea obtained from Rosmarinus officinalis L. collected in a sub‐humid area of Tunisia in order to assess whether the traditional rosemary tea preparation method could be considered as a reference method for rosemary's compounds extraction.

Methodology

Qualitative characterisation of Rosmarinus officinalis tea obtained after rosemary infusion in boiled water was determined by high performance liquid chromatography coupled with electrospray ionisation quadrupole time‐of‐flight mass spectrometry (HPLC‐ESI‐QTOF‐MS). Quantitative analysis relies on high performance liquid chromatography with diode array detector (HPLC‐DAD).

Results

Forty‐nine compounds belonging to six families, namely flavonoids, phenolic acids, phenolic terpenes, jasmonate, phenolic glycosides, and lignans were identified. To the best of the authors' knowledge eucommin A is characterised for the first time in rosemary. Rosmarinic acid (158.13 μg/g dried rosemary) was the main compound followed then by feruloylnepitrin (100.87 μg/g) and luteolin‐3′‐O‐(2″‐O‐acetyl)‐β‐d ‐glucuronide (44.04 μg/g). Among quantified compounds, luteolin‐7‐O‐rutinoside was the compound with the lowest concentration.

Conclusion

The infusion method allows several polyphenols present in rosemary tea to be extracted, therefore it could be a reference method for rosemary's compounds extraction. Moreover, traditional Tunisian Rosmarinus officinalis tea consumption is of interest for its rich phenolic content. Copyright © 2017 John Wiley & Sons, Ltd.     

Structural Characterization of <Emphasis Type="Italic">β</Emphasis>-glucans of <Emphasis Type="Italic">Agaricus brasiliensis</Emphasis> in Different Stages of Fruiting Body Maturity and their Use in Nutraceutical Products

β-Glucans of Agaricus brasiliensis fruiting bodies in different stages of maturity were isolated and characterized by FTIR and NMR. These fractions had greater amount of (1→6)-β-glucan and the (1→3)-β-glucan increased with fruiting bodies maturation. Yields of β-glucans increased from 42 mg β-glucans g⁻¹ fruiting bodies (dry wt) in immature stage to 43 mg g⁻¹ in mature stage with immature spores, and decreased to 40 mg g⁻¹ in mature stage with spore maturation. Mature fruiting bodies, which included these glucans, have potential therapeutical benefits for use in nutraceutical products.     

Antiproliferative Effect and Cell Cycle Alterations Induced by Salvia officinalis Essential Oil and Its Three Main Components in Human Colon Cancer Cell Lines

Colon cancer is one of the most common human malignancies, and chemotherapy cannot yet prevent recurrence in all patients. Essential oils are phytocomplexes with antiproliferative properties. In this study, we elucidated the antiproliferative properties and the effect on cell cycle progression of Sicilian Salvia officinalis essential oil and its three main compounds, α‐thujone, 1,8‐cineole (eucalyptol) and camphor, on three human colon cancer cell lines. The essential oil was obtained by hydrodistillation and analyzed by gas chromatography. Cell proliferation was evaluated by MTT assay, and the cell cycle distribution was determined by flow cytometry. Thirty‐four compounds were identified in the tested essential oil. Growth inhibition was observed after 72 h, with an impact on cell cycle progression and no effect on the viability of normal colonic epithelial cells. The study shows that S. officinalis essential oil and its three main components have an in vitro antiproliferative effect on colon cancer cells.     

基于Pacbio第三代测序技术的厚朴基因组测序分析

厚朴为著名的传统药用植物,归于木兰科、木兰属,于我国广泛种植,其树皮、根皮、枝皮、叶片、花、果实均能入药或食用。为获取厚朴全基因组序列信息,该文以厚朴叶片DNA为材料,采用Pacbio Sequel第三代测序技术构建厚朴全基因组数据库,并利用生物信息学方法对获得的核苷酸序列进行组装、功能注释以及进化分析研究。结果表明:(1)原始测序数据过滤后获得140.91 Gb三代数据,Read N50约为13 784bp,经过组装得到厚朴基因组大小为1.68 Gb,Contig N50约为222 069 bp,单拷贝基因完整性为81.0%。(2)组装后的序列通过与NR、KOG、KEGG等功能数据库比对,共有98.40%的基因得到了功能注释,其中KOG功能注释结果发现厚朴的蛋白功能主要集中在一般功能预测、翻译后修饰、蛋白质转换、伴侣以及信号转导机制; GO功能分类表明厚朴的基因集中在细胞组分及生物学过程; KEGG分析发现厚朴参与代谢通路的基因占主要地位。(3)通过与葡萄、拟南芥、水稻、杨树、银杏、无油樟、茶树及牛樟基因组的比对分析,发现厚朴23 424个基因中有20 801个基因可以分类到12 129个家族,其中有515个基因家族为厚朴所特有,而厚朴与牛樟(樟科)亲缘关系较近,两者的分化时间约在122.5百万年前(mya)。该研究首次利用第三代测序技术对厚朴全基因组解析,有利于对其进一步进行深入的开发与利用,也为研究其他药用植物全基因组奠定了基础。     

Inhibition of quorum sensing in Chromobacterium violaceum by pigments extracted from Auricularia auricular

Aims: This study aimed to search for a novel quorum‐sensing inhibitor from some fungi and analyse its inhibitory activity. Methods and Results: Chromobacterium violaceum CV026, a double mini‐Tn5 mutant, was used as an indicator to monitor quorum‐sensing inhibition. Auricularia auricular pigments from fruiting bodies were extracted using hydrochloric acid as an infusion, dissolved in alkaline dimethylsulfoxide (DMSO), sterilized by filtration through a 0·22‐μm membrane filter and added to C. violaceum CV026 cultures. Inhibitory activity was measured by quantifying violacein production using a microplate reader. The results have revealed that the alkaline DMSO‐soluble pigments significantly reduced violacein production in a concentration‐dependent manner, a quorum‐sensing‐regulated behaviour in C. violaceum. Conclusions: Auricularia auricular pigments can inhibit bacterial quorum sensing. Significance and Impact of the Study: The results suggest the bioactive constituents from edible and medicinal fungi could interfere with bacterial quorum‐sensing system, regulate its associate functions and prevent bacterial pathogenesis. Further studies were in process in our laboratory to isolate specific compounds from A. auricular pigments, evaluate them as quorum‐sensing inhibitors and analyse the exact mechanism of action.     

Kin Discrimination in Dictyostelium Social Amoebae

Evolved cooperation is stable only when the benefactor is compensated, either directly or through its relatives. Social amoebae cooperate by forming a mobile multicellular body in which, about 20% of participants ultimately die to form a stalk. This benefits the remaining individuals that become hardy spores at the top of the stalk, together making up the fruiting body. In studied species with stalked migration, P. violaceum, D. purpureum, and D. giganteum, sorting based on clone identity occurs in laboratory mixes, maintaining high relatedness within the fruiting bodies. D. discoideum has unstalked migration, where cell fate is not fixed until the slug forms a fruiting body. Laboratory mixes show some degree of both spatial and genotype‐based sorting, yet most laboratory fruiting bodies remain chimeric. However, wild fruiting bodies are made up mostly of clonemates. A genetic mechanism for sorting is likely to be cell adhesion genes tgrB1 and tgrC1, which bind to each other. They are highly variable, as expected for a kin discrimination gene. It is a puzzle that these genes do not cause stronger discrimination between mixed wild clones, but laboratory conditions or strong sorting early in the social stage diminished by later slug fusion could be explanations.