Phytochemistry (v.114, #C)

Editorial for the special issue on Ganoderma by R. Russell M. Paterson; Nelson Lima (5-6).

Global diversity of the Ganoderma lucidum complex (Ganodermataceae, Polyporales) inferred from morphology and multilocus phylogeny by Li-Wei Zhou; Yun Cao; Sheng-Hua Wu; Josef Vlasák; De-Wei Li; Meng-Jie Li; Yu-Cheng Dai (7-15).
Ganoderma lingzhi is the most widely cultivated medicinal species within the Ganoderma lucidum complex, within which 13 species have been recognized from morphology and phylogeny.Display OmittedSpecies of the Ganoderma lucidum complex are used in many types of health products. However, the taxonomy of this complex has long been chaotic, thus limiting its uses. In the present study, 32 collections of the complex from Asia, Europe and North America were analyzed from both morphological and molecular phylogenetic perspectives. The combined dataset, including an outgroup, comprised 33 ITS, 24 tef1α, 24 rpb1 and 21 rpb2 sequences, of which 19 ITS, 20 tef1α, 20 rpb1 and 17 rpb2 sequences were newly generated. A total of 13 species of the complex were recovered in the multilocus phylogeny. These 13 species were not strongly supported as a single monophyletic lineage, and were further grouped into three lineages that cannot be defined by their geographic distributions. Clade A comprised Ganoderma curtisii, Ganoderma flexipes, Ganoderma lingzhi, Ganoderma multipileum, Ganoderma resinaceum, Ganoderma sessile, Ganoderma sichuanense and Ganoderma tropicum, Clade B comprised G. lucidum, Ganoderma oregonense and Ganoderma tsugae, and Clade C comprised Ganoderma boninense and Ganoderma zonatum. A dichotomous key to the 13 species is provided, and their key morphological characters from context, pores, cuticle cells and basidiospores are presented in a table. The taxonomic positions of these species are briefly discussed. Noteworthy, the epitypification of G. sichuanense is rejected.
Keywords: Taxonomy; Polypore; ITS; tef1α; rpb1; rpb2;

The nomenclature of Ganoderma used as a Chinese medicine is debated. A group of researchers could not amplify the DNA of type specimens and concluded the DNA was degraded irreparably. New topotypes were used as the type specimens which was premature. The use of internal amplification controls is recommended to determine if other factors were involved as alternative explanations.

Genome analysis of medicinal Ganoderma spp. with plant-pathogenic and saprotrophic life-styles by Ursula Kües; David R. Nelson; Chang Liu; Guo-Jun Yu; Jianhui Zhang; Jianqin Li; Xin-Cun Wang; Hui Sun (18-37).
This review carries out a detailed comparison of the nuclear genomes, mitochondrial genomes and transcriptomes from several Ganoderma species. Genes involved in biosynthetic pathways such as CYP450 genes and in cellular development such as matA and matB genes are characterized and compared in detail.Display Omitted Ganoderma is a fungal genus belonging to the Ganodermataceae family and Polyporales order. Plant-pathogenic species in this genus can cause severe diseases (stem, butt, and root rot) in economically important trees and perennial crops, especially in tropical countries. Ganoderma species are white rot fungi and have ecological importance in the breakdown of woody plants for nutrient mobilization. They possess effective machineries of lignocellulose-decomposing enzymes useful for bioenergy production and bioremediation. In addition, the genus contains many important species that produce pharmacologically active compounds used in health food and medicine. With the rapid adoption of next-generation DNA sequencing technologies, whole genome sequencing and systematic transcriptome analyses become affordable approaches to identify an organism’s genes. In the last few years, numerous projects have been initiated to identify the genetic contents of several Ganoderma species, particularly in different strains of Ganoderma lucidum. In November 2013, eleven whole genome sequencing projects for Ganoderma species were registered in international databases, three of which were already completed with genomes being assembled to high quality. In addition to the nuclear genome, two mitochondrial genomes for Ganoderma species have also been reported. Complementing genome analysis, four transcriptome studies on various developmental stages of Ganoderma species have been performed. Information obtained from these studies has laid the foundation for the identification of genes involved in biological pathways that are critical for understanding the biology of Ganoderma, such as the mechanism of pathogenesis, the biosynthesis of active components, life cycle and cellular development, etc. With abundant genetic information becoming available, a few centralized resources have been established to disseminate the knowledge and integrate relevant data to support comparative genomic analyses of Ganoderma species. The current review carries out a detailed comparison of the nuclear genomes, mitochondrial genomes and transcriptomes from several Ganoderma species. Genes involved in biosynthetic pathways such as CYP450 genes and in cellular development such as matA and matB genes are characterized and compared in detail, as examples to demonstrate the usefulness of comparative genomic analyses for the identification of critical genes. Resources needed for future data integration and exploitation are also discussed.
Keywords: Ganoderma; Whole genome sequencing; Mitochondrial genome; Transcriptome; CYP450; matA; matB;

Chemical features of Ganoderma polysaccharides with antioxidant, antitumor and antimicrobial activities by Isabel C.F.R. Ferreira; Sandrina A. Heleno; Filipa S. Reis; Dejan Stojkovic; Maria João R.P. Queiroz; M. Helena Vasconcelos; Marina Sokovic (38-55).
This review aims to contribute to the knowledge on bioactivity (antioxidant, antitumor and antimicrobial properties) of polysaccharides, glycoproteins and polysaccharidic extracts obtained from Ganoderma species. The chemical features were analyzed and related with the correspondent bioactivity.Display Omitted Ganoderma genus comprises one of the most commonly studied species worldwide, Ganoderma lucidum. However, other Ganoderma species have been also reported as important sources of bioactive compounds. Polysaccharides are important contributors to the medicinal properties reported for Ganoderma species, as demonstrated by the numerous publications, including reviews, on this matter. Yet, what are the chemical features of Ganoderma polysaccharides that have bioactivity? In the present manuscript, the chemical features of Ganoderma polysaccharides with reported antioxidant, antitumor and antimicrobial activities (the most studied worldwide) are analyzed in detail. The composition of sugars (homo- versus hetero-glucans and other polysaccharides), type of glycosidic linkages, branching patterns, and linkage to proteins are discussed. Methods for extraction, isolation and identification are evaluated and, finally, the bioactivity of polysaccharidic extracts and purified compounds are discussed. The integration of data allows deduction of structure–activity relationships and gives clues to the chemical aspects involved in Ganoderma bioactivity.
Keywords: Ganoderma; Polysaccharides; Chemical features; Bioactivity; Structure–activity;

From 2000 years of Ganoderma lucidum to recent developments in nutraceuticals by Karen S. Bishop; Chi H.J. Kao; Yuanye Xu; Marcus P. Glucina; R. Russell M. Paterson; Lynnette R. Ferguson (56-65).
A review. Ganoderma lucidum has been revered as a medicinal mushroom to treat assorted diseases and prolong life. Nowadays it is a multi-billion dollar industry.Display OmittedMedicinal mushrooms have been used for centuries as nutraceuticals to improve health and to treat numerous chronic and infectious diseases. One such mushroom is Ganoderma lucidum, commonly known as Lingzhi, a species revered as a medicinal mushroom for treating assorted diseases and prolonging life. The fungus is found in diverse locations, and this may have contributed to confusion regarding the correct taxonomic classification of the genus Ganoderma. G. lucidum was first used to name a specimen found in England and thereafter was naively applied to a different Ganoderma species found in Asia, commonly known as Chinese Lingzhi. Despite the taxonomic confusion, which has largely been uncorrected, the popularity of Lingzhi has escalated across the globe. The current taxonomic situation is now discussed accurately in this Special Issue on Ganoderma. Today it is a multi-billion dollar industry wherein Lingzhi is cultivated or collected from the wild and consumed as a tea, in alcoholic beverages, and as a nutraceutical to confer numerous health benefits. Consumption of nutraceuticals has grown in popularity, and it is becoming increasingly important that active ingredients be identified and that suppliers make substantiated health claims about their products. The objective of this article is to present a review of G. lucidum over the past 2000 years from prized ancient “herbal” remedy to its use in nutraceuticals and to the establishment of a 2.5 billion $ (US) industry.
Keywords: Medicinal mushrooms; Ganoderma lucidum; Ganodermataceae; Lingzhi; Taxonomy; History; Health benefits; Nutraceuticals;

Secondary metabolites from Ganoderma by Sabulal Baby; Anil John Johnson; Balaji Govindan (66-101).
Ganoderma is a genus of medicinal mushrooms. Phytochemical studies led to the isolation of over 380 terpenoids (ganoderic acids, lucidenic acids, meroterpenoids) from genus Ganoderma. More than 30 steroidal compounds were also isolated from the genus. Terpenoids, steroids (and polysaccharides) isolated from Ganoderma showed significant biological activities.Display Omitted Ganoderma is a genus of medicinal mushrooms. This review deals with secondary metabolites isolated from Ganoderma and their biological significance. Phytochemical studies over the last 40 years led to the isolation of 431 secondary metabolites from various Ganoderma species. The major secondary compounds isolated are (a) C30 lanostanes (ganoderic acids), (b) C30 lanostanes (aldehydes, alcohols, esters, glycosides, lactones, ketones), (c) C27 lanostanes (lucidenic acids), (d) C27 lanostanes (alcohols, lactones, esters), (e) C24, C25 lanostanes (f) C30 pentacyclic triterpenes, (g) meroterpenoids, (h) farnesyl hydroquinones (meroterpenoids), (i) C15 sesquiterpenoids, (j) steroids, (k) alkaloids, (l) prenyl hydroquinone (m) benzofurans, (n) benzopyran-4-one derivatives and (o) benzenoid derivatives. Ganoderma lucidum is the species extensively studied for its secondary metabolites and biological activities. Ganoderma applanatum, Ganoderma colossum, Ganoderma sinense, Ganoderma cochlear, Ganoderma tsugae, Ganoderma amboinense, Ganoderma orbiforme, Ganoderma resinaceum, Ganoderma hainanense, Ganoderma concinna, Ganoderma pfeifferi, Ganoderma neo-japonicum, Ganoderma tropicum, Ganoderma australe, Ganoderma carnosum, Ganoderma fornicatum, Ganoderma lipsiense (synonym G. applanatum), Ganoderma mastoporum, Ganoderma theaecolum, Ganoderma boninense, Ganoderma capense and Ganoderma annulare are the other Ganoderma species subjected to phytochemical studies. Further phytochemical studies on Ganoderma could lead to the discovery of hitherto unknown biologically active secondary metabolites.
Keywords: Ganoderma; Ganodermataceae; Lanostane type triterpenoids; Ganoderic acids; Lucidenic acids; Meroterpenoids; Steroids; Volatiles; Biological activities;

Ganoderma pfeifferi – A European relative of Ganoderma lucidum by Ulrike Lindequist; Wolf-Dieter Jülich; Sabine Witt (102-108).
Ganoderma pfeifferi contains farnesylhydroquinones named ganomycins and several triterpenoid compounds. Extracts and some of the isolated compounds possess antibacterial, antiviral and other biological activities.Display OmittedIn contrast to well-studied and broadly used Ganoderma species, such as Ganoderma lucidum and Ganoderma applanatum, knowledge regarding Ganoderma pfeifferi is very limited. Herein is an overview of the phytochemistry, biological activities and possible applications of this mushroom species. In addition to triterpenoids and polysaccharides, G. pfeifferi contains unique sesquiterpenoids and other small molecular weight compounds. Some of these compounds exhibit remarkable antimicrobial activities in vitro and in vivo against multi-resistant bacteria, such as MRSA. Antiviral properties, UV-protection abilities and other activities are also known. Potential issues arising from the conversion of research results into practical applications are discussed.
Keywords: Ganoderma pfeifferi; Ganodermataceae; Farnesylhydroquinones; Triterpenoids; Antibacterial; Antiviral; Keratinocytes;

Anti-diabetic effects of Ganoderma lucidum by Haou-Tzong Ma; Jung-Feng Hsieh; Shui-Tein Chen (109-113).
This review summarizes the anti-diabetic effects of Ganoderma lucidum. Four types of molecules, proteins, proteoglycans, polysaccharides and triterpenoids were shown to regulate multiple pathways in the performance of anti-diabetic activities.Display Omitted Ganoderma lucidum is a white rot fungus widely used as a tonic for the promotion of longevity and health. Extracts of G. lucidum have been recognized as an alternative adjuvant treatment for diabetes. Among the many biologically active constituents of G. lucidum, polysaccharides, proteoglycans, proteins and triterpenoids have been shown to have hypoglycemic effects. G. lucidum polysaccharides have been reported to have hypoglycemic activity by increasing plasma insulin levels and decreasing plasma sugar levels in mice. Protein tyrosine phosphatase 1B is a promising therapeutic target in diabetes, and G. lucidum proteoglycan can inhibit this enzyme in vitro. Moreover, G. lucidum triterpenoids were shown to have inhibitory activity on aldose reductase and α-glucosidase that can suppress postprandial hyperglycemia. In addition, a protein Ling Zhi-8 extracted from G. lucidum significantly decreased lymphocyte infiltration and increased the antibody detection of insulin in diabetic mice. This review summarizes most of the research about the hypoglycemic action effects of polysaccharides, proteoglycans, proteins and tritrerpenoids from G. lucidum as a guide for future research.
Keywords: Ganoderma lucidum; Ganodermataceae; Triterpenoids; Polysaccharides; Diabetes;

Accessing biological actions of Ganoderma secondary metabolites by in silico profiling by Ulrike Grienke; Teresa Kaserer; Florian Pfluger; Christina E. Mair; Thierry Langer; Daniela Schuster; Judith M. Rollinger (114-124).
A multi-conformational 3D molecular structure database comprising 279 Ganoderma constituents was generated and subjected to pharmacophore-based virtual parallel screening using 529 pharmacophore models covering targets related to viral diseases and the metabolic syndrome. Hit lists were evaluated by comparison with activities reported in literature.Display OmittedThe species complex around the medicinal fungus Ganoderma lucidum Karst. (Ganodermataceae) is widely known in traditional medicines, as well as in modern applications such as functional food or nutraceuticals. A considerable number of publications reflects its abundance and variety in biological actions either provoked by primary metabolites, such as polysaccharides, or secondary metabolites, such as lanostane-type triterpenes. However, due to this remarkable amount of information, a rationalization of the individual Ganoderma constituents to biological actions on a molecular level is quite challenging. To overcome this issue, a database was generated containing meta-information, i.e., chemical structures and biological actions of hitherto identified Ganoderma constituents (279). This was followed by a computational approach subjecting this 3D multi-conformational molecular dataset to in silico parallel screening against an in-house collection of validated structure- and ligand-based 3D pharmacophore models. The predictive power of the evaluated in silico tools and hints from traditional application fields served as criteria for the model selection. Thus, the focus was laid on representative druggable targets in the field of viral infections (5) and diseases related to the metabolic syndrome (22). The results obtained from this in silico approach were compared to bioactivity data available from the literature. 89 and 197 Ganoderma compounds were predicted as ligands of at least one of the selected pharmacological targets in the antiviral and the metabolic syndrome screening, respectively. Among them only a minority of individual compounds (around 10%) has ever been investigated on these targets or for the associated biological activity. Accordingly, this study discloses putative ligand target interactions for a plethora of Ganoderma constituents in the empirically manifested field of viral diseases and metabolic syndrome which serve as a basis for future applications to access yet undiscovered biological actions of Ganoderma secondary metabolites on a molecular level.
Keywords: Ganoderma lucidum; Ganodermataceae; Pharmacophore profiling; Virtual screening; Triterpenes; Antiviral targets; Metabolic syndrome;

Anti-inflammatory activity on mice of extract of Ganoderma lucidum grown on rice via modulation of MAPK and NF-κB pathways by Md. Abul Hasnat; Mehnaz Pervin; Kyu Min Cha; Si Kwan Kim; Beong Ou Lim (125-136).
Ganoderma lucidum exerts anti-inflammatory activity with an enhanced colitis safety profile via simultaneous inhibition of phosphorylation of MAPKs and activation of NF-κB.Display Omitted Ganoderma lucidum is a popular medicinal mushroom with anti-inflammatory potential. In the present study, the aim was to determine the anti-inflammatory effect and mode of action of G. lucidum grown on germinated brown rice (GLBR) in a mouse model of colitis. It was shown that GLBR suppressed the production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharide (LPS)-stimulated macrophages and decreased the expression of COX-2, TNF-α, iNOS, IL-1β, IL-6, and IL-10 mRNAs. GLBR also inhibited activation of p38, ERK, JNK, MAPKs, and nuclear factor kappa-B (NF-κB). In a mouse model of colitis, colonic mucosal injury was evaluated using macroscopic, biochemical, and histopathological testing. Disease activity index (DAI), macroscopic score, and histological score significantly decreased upon GLBR treatment. Moreover, immunofluorescence studies indicated that DSS activates nuclear translocation of NF-κB in colon tissue, which is attenuated by GLBR extract. These findings suggest that GLBR is protective against colitis via inhibition of MAPK phosphorylation and NF-κB activation.
Keywords: Ganoderma lucidum; Ganodermataceae; Rice; Inflammatory bowel disease; Nuclear factor-kappa B; Mitogen-activated protein kinase;

Lanostane triterpenoids from Ganoderma hainanense J. D. Zhao by XingRong Peng; JieQing Liu; JianJun Xia; CuiFang Wang; XuYang Li; YuanYuan Deng; NiMan Bao; ZhiRun Zhang; MingHua Qiu (137-145).
Fourteen lanostane triterpenoids were identified from Ganoderma hainanense, of which one possesses a 29-norlanostane structure. X-ray single crystal diffraction analysis determined the absolute configuration of ganoderma acids from G. hainanense to be 25S.Display OmittedChemical investigation of the fruiting bodies of Ganoderma hainanense resulted in isolation of fourteen lanostane triterpenoids, including nine ganoderma acids and five ganoderma alcohols, together with five known compounds. Structural elucidation was determined using extensive spectroscopic technologies, Mosher’s method and X-ray single crystal diffraction. Three of the compounds showed inhibitory activities against HL-60, SMMC-7721, A-549 and MCF-7 cells with IC50 values of 15.0–40.0 μM.
Keywords: Ganoderma hainanense; Lanostane triterpenoids; X-ray diffraction; Mosher’s method; Cytotoxicity;

A reproducible analytical system based on the multi-component analysis of triterpene acids in Ganoderma lucidum by Juan Da; Chun-Ru Cheng; Shuai Yao; Hua-Li Long; Yan-Hong Wang; Ikhlas A. Khan; Yi-Feng Li; Qiu-Rong Wang; Lu-Ying Cai; Bao-Hong Jiang; Xuan Liu; Wan-Ying Wu; De-An Guo (146-154).
Ten triterpenes in Ganoderma lucidum were quantitatively determined using ultra performance liquid chromatography, with ganoderic acid A serving as the single reference standard. Stable system parameters were established.Display OmittedUltra-performance liquid chromatography (UPLC) and Single Standard for Determination of Multi-Components (SSDMC) are becoming increasingly important for quality control of medicinal herbs; this approach was developed for Ganoderma lucidum. Special attention was necessary for the appropriate selection of markers, for determining the reproducibility of the relative retention times (RRT), and for the accuracy of conversion factors (F). Finally, ten components were determined, with ganoderic acid A serving as single standard. Stable system parameters were established, and with successful resolution of those issues, this analytical method could be used more broadly.
Keywords: Ganoderma lucidum; Ganoderenic acids; Ganoderic acids; Quantitative determination; Ultra performance liquid chromatography;

Metabolites from the mushroom Ganoderma lingzhi as stimulators of neural stem cell proliferation by Yong-Ming Yan; Xin-Long Wang; Qi Luo; Li-Ping Jiang; Cui-Ping Yang; Bo Hou; Zhi-Li Zuo; Yong-Bin Chen; Yong-Xian Cheng (155-162).
Ten meroterpenoids with diverse ring systems and two known compounds were isolated from Ganoderma lingzhi. Several of them were found to promote proliferation of neural stem cells at 10 μM.Display Omitted Ganoderma lingzhi is a valuable, edible and medicinal fungus that has been widely used for the prevention and treatment of a broad range of diseases. In this study, spirolingzhines A–D, four meroterpenoids with a spiro[benzofuran-2,1′-cyclopentane] motif, lingzhines A–F, six meroterpenoids with diverse ring systems, along with two known compounds were isolated from the fruiting bodies of this fungus. The structures and stereochemistry of these substances were determined by using spectroscopic, X-ray crystallographic and computational methods. Chiral HPLC was used to separate (−)- and (+)-antipodes of seven of ten meroterpenoids, which were isolated from the fungus as racemic mixtures. Several of the metabolites were found to promote proliferation of neural stem cells (NSCs) and, as such, they constitute a class of NSC stimulators. The most potent member of this series, (−)-spirolingzhine A, was shown to affect NSC cell cycle progression using the 5-bromo-2-deoxyuridine (BrdU) incorporation assay.
Keywords: Ganoderma lingzhi; Ganodermataceae; Meroterpenoids; Neural stem cells; Neurodegenerative diseases;

(+)-Epogymnolactam, a novel autophagy inducer from mycelial culture of Gymnopus sp. by Shinya Mitsuhashi; Chihaya Shindo; Kengo Shigetomi; Toshizumi Miyamoto; Makoto Ubukata (163-167).
An autophagy inducer, named (+)-epogymnolactam, was discovered from the mycelial culture of Gymnopus sp., and exists as a tautomeric mixture. The major form in MeOH was (1R,5S)-4-butyl-4-hydroxy-3-aza-6-oxa-bicyclo[3.1.0]hexan-2-one.Display OmittedMushrooms, including Ganoderma lucidum, have been used as a potential source of therapeutic compounds, and an autophagy inducer would be useful for treatment of diverse diseases in human. Reported here is a full account of screening, isolation, structural determination, and biological activity of an autophagy inducer, (+)-epogymnolactam (1) from a mycelial culture of a Gymnopus sp. strain. Its structure was elucidated by HR-ESI-MS, NMR, and its plus sign by specific rotation. It exists as a tautomeric mixture of 1a and 1b in MeOH. The major tautomer of 1 is (1R,5S)-4-butyl-4-hydroxy-3-aza-6-oxa-bicyclo[3.1.0]hexan-2-one (1a), and the minor tautomeric form is (2R,3S)-3-pentanoyloxirane-2-carboxamide (1b).
Keywords: Mushroom; Gymnopus sp.; Isolation; Biologically active substance; Autophagy; (+)-Epogymnolactam; (1R,5S)-4-Butyl-4-hydroxy-3-aza-6-oxa-bicyclo[3.1.0]hexan-2-one; (2R,3S)-3-Pentanoyloxirane-2-carboxamide;

The molecular defence responses of oil palm to Ganoderma infection in basal stem rot are reviewed based on the transcript profiles of pathogenesis-related proteins in infected oil palms.Display OmittedBasal stem rot (BSR) of oil palm roots is due to the invasion of fungal mycelia of Ganoderma species which spreads to the bole of the stem. In addition to root contact, BSR can also spread by airborne basidiospores. These fungi are able to break down cell wall components including lignin. BSR not only decreases oil yield, it also causes the stands to collapse thus causing severe economic loss to the oil palm industry. The transmission and mode of action of Ganoderma, its interactions with oil palm as a hemibiotroph, and the molecular defence responses of oil palm to the infection of Ganoderma boninense in BSR are reviewed, based on the transcript profiles of infected oil palms. The knowledge gaps that need to be filled in oil palm–Ganoderma molecular interactions i.e. the associations of hypersensitive reaction (HR)-induced cell death and reactive oxygen species (ROS) kinetics to the susceptibility of oil palm to Ganoderma spp., the interactions of phytohormones (salicylate, jasmonate and ethylene) at early and late stages of BSR, and cell wall strengthening through increased production of guaiacyl (G)-type lignin, are also discussed.
Keywords: Ganoderma; Oil palm; Elaeis guineensis Jacq.; Basal stem rot; Molecular defence; Lignin;