Dissertations / Theses on the topic 'Ganoderma'

Ganoderma species are one of the most widely researched fungi because of their reported potent bioactive properties. Although there is much information related to American, European and Asian isolates, little research has been conducted on Australian Ganoderma isolates. Ganoderma may only be imported into Australia under strict quarantine conditions, therefore, the isolation of a native strain that possesses bioactivity may be industrially and commercially significant. Three Australian species of this wood-decomposing fungus were isolated in northern Queensland. In this study, they have been identified as three separate species. Further, they have been studied to determine their optimal growth conditions in liquid culture and assessed for their antibacterial properties. Phylogeny inferred from the Internal Transcribed Spacer Regions (ITS) from the DNA sequences resolved the three Australian Ganoderma species into separate clades. Two isolates were identified to be isolates of Ganoderma cupreum (H1) and Ganoderma weberianum (H2). The third isolate could only be identified to the genus level, Ganoderma species, due to the lack of informative data that could be used for comparison. The effects of short term and long term storage on the viability of the fungi were investigated on agar plates, agar slants and balsa wood at varying temperatures ranging from 10 to 45�C. The most appropriate storage conditions were determined to be �80�C on balsa wood chips for periods of up to 2 years without subculture, and on agar slants at 4�C for up to a maximum of eight weeks. Light was observed to be detrimental to the survival of Ganoderma H1 and Ganoderma H2 during storage. Growth trials using potato dextrose agar plates determined the optimal temperature and pH for mycelial growth to be 30�C and a pH of 6, for all isolates. Subsequent growth trials in liquid media found that glucose, as the carbohydrate source, supported the greatest mycelial growth of Ganoderma H1 and Ganoderma H2 and that galactose and fructose supported the greatest growth of Ganoderma H3. Abstract ii Aqueous (hot water) and organic (hexane (HEX), dichloromethane (DCM), ethyl acetate (EtOAc), methanol (MeOH)) extracts from the liquid cultivated mycelium were assessed for their antibacterial activity using disc diffusion assays. Extracts from the mycelium of Ganoderma H1 exhibited activity against a greater number of Gram positive bacteria than those from Ganoderma H2 and H3. Subsequent studies on the DCM and EtOAc extracts from Ganoderma H1 determined the MIC and MBC against a number of Gram positive bacteria, including Bacillus cereus, B. subtilis, Enterococcus faecalis, Streptococcus pyogenes, Staphylococcus aureus, S. epidermidis and Listeria monocytogenes, as well as Clostridium species, including Clostridium perfringens, C. sporogenes and C. difficile, and some methicillin resistant Staphylococcus aureus (MRSA) strains. Time course growth assays confirmed that the DCM and EtOAc extracts predominantly exhibited bactericidal activity. Finally, the active compounds were determined to be terpenoid in structure with some phenolic groups attached.

Ganoderma lucidum and G. tsugae are two members of the G. lucidum complex. The authenticity of the two wood-rotting species was demonstrated by comparative studies. Ganoderma lucidum is restricted to hardwoods. Its "smooth" walled basidiospores were characterized by narrow, numerous inter-wall pillars. Isolates of G. lucidum produced chlamydospores in culture and had an average growth of 7.8 mm/da at their optimum temperature range of 30-34 C. Ganoderma tsugae is restricted to conifers. Its basidiospores were "rough" walled and had broad inter-wall pillars. Isolates of G. tsugae did not produce chlamydospores in culture and had an average growth of 2.1 mm/da at the optimum temperature range of 20-25 C. Mating systems were determined for both species as heterothallic and tetrapolar. Interspecific matings of homokaryons were incompatible. Homokaryons of a European G. resinaceum isolate were interfertile with homokaryons from North American collections of G. lucidum. The ability of G. lucidum and G. tsugae to decay wood in vitro was studied using the following woods in agar block decay chambers: grape, oak, mesquite, white fir, and Douglas-fir. Grape wood lost the most weight while mesquite the least. G. lucidum isolates generally caused greater weight loss of all woods than did G. tsugae isolates. Both Ganoderma species caused simultaneous decay in all woods. However, chemical analyses of the decayed blocks indicated that selective delignification by both species also occurred in grape and white fir blocks but not in oak or Douglas-fir blocks. Scanning electron microscopy demonstrated various stages of selective delignification and simultaneous decay of all woods tested. Isolates of Ganoderma lucidum infected Dog Ridge variety grape plants, grown in the greenhouse, from below-ground wood block inoculations. Twenty-four plants were inoculated: one plant died and 4 other plants declined. After 24 months reisolations yielded only G. lucidum from the five declining plants, demonstrating pathogenicity. The fungus developed in the heartwood and, in later stages, invaded the sapwood. Infected plants developed water stress symptoms with leaves wilting, yellowing, and dying. Field grape plants inoculated with the fungus developed decay columns as large as 42 cm in 17 mons. Decay was limited to the heartwood; no foliar symptoms occurred.

En premier lieu, notre travail résume l’état des connaissances sur les Ganodermataceae des Antilles françaises, en se basant sur l’analyse exhaustive de la littérature et sur les récoltes personnelles que nous avons effectuées au cours de 7 missions sur le terrain, en Guadeloupe et en Martinique (2003- 2008). Soixante quatre collections ont été examinées, ainsi que plusieurs types d’espèces décrites des Néotropiques (Ganoderma dussii Pat. , G. Pulverulentum Murrill, G. Subfornicatum Murrill, G. Tuberculosum Murrill). Au total, quinze taxons ont été identifiés dans les Antilles françaises. Les caractères morphologiques de chaque espèce ont fait l’objet d’une description méticuleuse, d’une illustration sur planche ainsi que d’une discussion. De plus, chacune de nos hypothèses portées sur l’identité des espèces s’appuie sur des résultats phylogénétiques. Les espèces suivantes sont citées pour la première fois aux Antilles : anoderma amazonense Weir, Ganoderma flaviporum (Murill) Sacc. & Trotter, Ganoderma subamboinense (P. Hennings) Bazzalo & Wright, Ganoderma tuberculosum Murril. En second lieu, nos recherches ont porté sur la comparaison des activités antiprolifératives de différents extraits méthanoliques de Ganodermataceae Donk récoltés aux Antilles et Guyane françaises et de deux espèces de référence : Ganoderma lucidum européen et ‘Ganoderma lucidum’ asiatique, sur cellules cancéreuses humaines de type prostatique (PC-3), mammaires (MCF- 7) et colorectal (HT-29). Nous avons démontré que certains ganodermes néotropicaux tel le Ganoderma tuberculosum pouvaient aussi inhiber la croissance des cellules cancéreuses de la même façon, sinon plus, que le Ganoderma lucidum utilisé en médecine traditionnelle asiatique et le Ganoderma lucidum européen. Les acides ganodériques A, DM et F, pour lesquels une activité anticancéreuse a été démontrée, n’ont pas été retrouvés dans l’extrait méthanolique de Ganoderma tuberculosum. D’un autre côté, l’extrait de cette espèce contient un composé majoritaire correspondant 256 à un acide ganodérique jusqu’ici jamais décrit : GA FWI. Celui-ci n’a pas montré d’activité antiproliférative significative sur cellules cancéreuses de type PC-3, MCF-7 et HT29. De plus, ce travail a montré que l’activité antiproliférative du G. Lucidum européen est similaire à celle du G. Lucidum traditionnellement utilisé en médecine asiatique excepté pour HT29 pour lesquels aucune activité n’a été démontrée pour l’espèce asiatique. Enfin, notre étude illustre l'importance des investigations taxinomiques fiables, avec des méthodes traditionnelles et moléculaires, afin d'identifier clairement les espèces contenant les molécules spécifiques bioactives.

Ganoderma lucidum is a popular herbal medicine used in China to promote health. Modern studies have disclosed that the active ingredients of Ganoderma can exhibit several effects, including antitumor effects and immunomodulation. The present study evaluated the antitumor effects of self-prepared Ganoderma extracts and spores oil, and investigated the possible underlying mechanisms by observing the effects of the extracts and oil on topoisomerases and the cell cycle. The results showed that Ganoderma extracts and spores oil presented dose-dependent inhibitory effects on tumor cells. The half maximal inhibitory concentration (IC50) values of Ganoderma extracts on HL60, K562 and SGC-7901 cells for 24 h were 0.44, 0.39 and 0.90 mg/ml, respectively; for Ganoderma spores oil, the IC50 values were 1.13, 2.27 and 6.29 mg/ml, respectively. In the in vivo study, the inhibitory rates of Ganoderma extracts (4 g/kg/d, intragastrically) on S180 and H22 cells were 39.1 and 44.6%, respectively, and for Ganoderma spores oil (1.2 g/kg/d, intragastrically) the inhibitory rates were 30.9 and 44.9%, respectively. Ganoderma extracts and spores oil inhibited the activities of topoisomerase I and II. Ganoderma spores oil was shown block the cell cycle at the transition between the G1 and S phases and induce a marked decrease in cyclin D1 levels in K562 cells, with no significant change in cyclin E level. These results suggest that the Ganoderma extracts and spores oil possessed antitumor effects in the in vitro and in vivo studies. The antitumor mechanisms of the extracts and spores oil were associated with inhibitory effects on topoisomerase I and II activities, and for Ganoderma spores oil, the antitumor effects may also be associated with decreased cyclin D1 levels, thus inducing G1 arrest in the cell cycle.

La compréhension de la structure génétique et de la dynamique des populations, ainsi que les mécanismes moléculaires régissant les interactions entre l’hôte et le pathogène sont des éléments clefs pour la gestion des maladies. Au cours de cette étude nous avons cherché à développer dans un premier temps des marqueurs microsatellites à partir de données issues du séquençage d’un isolat de Ganoderma boninense. Ces marqueurs microsatellites nous ont permis d’étudier la structuration génétique et l’histoire démographique de G. boninense. Le génotypage d’un sous-échantillonnage issus de missions de récolte en Malaisie et à Sumatra nous a permis de mettre en évidence deux groupes principaux au sein de l’Asie du Sud-Est. Nous avons ensuite concentré notre attention sur la région regroupant la Malaisie péninsulaire et l’île de Sumatra, zone historique de développement de la culture industrielle du palmier à huile et d’apparition de la pourriture basale du stipe due à G. boninense, qui semble former une seule population. Nous avons examiné à une échelle géographique plus restreinte cette zone géographique afin de mettre en évidence une potentielle sous structure au sein de cette population. En testant l’effet du fond génétique du palmier d’origine de chaque individu, du nombre de génération de palmier précédent le moment de la récolte de l’individu, ou de la distance géographique sur une possible sous-structure des individus au sein de la zone historique, nous avons mis en évidence qu’aucune sous-structure génétique n’émergeait. En revanche, La comparaison de plusieurs scénarios d’évolution démographique a permis de mettre en évidence un phénomène d’expansion très ancien bien antérieur au début du développement de la culture industrielle du palmier à huile. Pour finir, la comparaison des données transcriptomiques entre des isolats agressifs et non agressifs a permis de souligner la présence d’une centaine de gènes différentiellement exprimés possédant une annotation fonctionnelle. Les résultats de ces deux approches pourront permettre une meilleure gestion de la maladie ainsi que l’amélioration des programmes développement et de gestion des résistances
The understanding of genetic structuration and population dynamic, as well as the molecular mechanisms ruling host/pathogen interaction, are key elements for disease management. During this study, as first step, we were looking to develop microsatellites markers from genomic data obtained from sequencing of a Ganoderma boninense pure strain. Those markers allowed studying genetic structuration and demographic history of G. boninense. Genotyping of a subset of samples from sampling mission in Malaysia and Sumatra have highlighted two main groups in South-East Asia. Then, we focused on a region gathering peninsular Malaysia and Sumatra together, as it is both historical region of industrial oil palm culture development and the first region of basal stem rot observation caused by G. boninense, and to appear as a single population. We examine this region at a lower scale in order to highlight a potential genetic substructure in this population. We tested for effect of genetic tree background of each sample, number of planting generation before sampling and geographical distance between sample in order to observe a potential correlation between genetic substructure and one of those factors. As no correlation appeared, we concluded that this population does not have a genetic substructure. On the other hand, the comparison between several demographic evolution scenarios have shown a strong support for a past expansion event further back in time from the beginning of industrial oil palm culture development. To conclude, the exploration of transcriptomic data between strains owning aggressive or non-aggressive profile showed the differential expression of a hundred genes owning a functional annotation. Results from both approaches will allow the development of better disease management and a better resistance selection and management program

Oil palm is an important crop to the economy of Malaysia and Indonesia. There have been considerable efforts to improve crop quality of crop to meet the growing demands for edible oil in the world. However, the threat of pests and diseases in Malaysian oil palm has increasingly challenged the production of crude palm oil, sometimes to the point of driving plantations to closure and conversion of estate land for commercial development. The most devastating disease in oil palm is basal stem rot (BSR), caused by the root-rot pathogen Ganoderma boninense. The disease has been observed in both young and mature oil palm and in both inland and coastal plantations. Epidemiology of BSR was explained through infection by spores and by mycelium from previously infected oil palm and coconut stands, and the presence of G. boninense in the basal stem and soil surrounding infected palms. The life cycle of G. boninense could extend to years as the fungus can remain as resting structures in palm tissues and as recalcitrant spores spread by wind and rain splash in estates. Shade house trials have been successful in producing artificially infected Ganoderma-oil palm BSR symptoms. However, the current method uses oil palm seedlings of three to 12 months and involves inoculation with G. boninense that has pre-colonised a rubber wood block for one month. This method requires a minimum of six months to observe BSR-like symptoms, a time consuming effort. The aim of this work was to develop an efficient artificial infection assay that uses clonal oil palm plantlets as hosts for BSR disease through the inoculation of G. boninense isolate GBLS. The experiment was set up with treatments of T1: non-treatment control, T2: wounded plant control and T3: wounded and GBLS-infected plants. During the incubation period of 42 days, T3 plants consistently showed significant stunting (5.18% and 13.41% shorter than T1 and T2, respectively) and loss of weight (57.58% and 61.00% lighter than T1 and T2, respectively). The T3 plants also had significantly thinner leaves (38.70% and 37.71% narrower than T1 and T2, respectively) and lower chlorophyll contents (42.95% and 64.88% lower SPAD readings than T1 and T2, respectively). Disease severity on the T3 plants was 100% by 6 weeks, indicating death of oil palms. The quantity of GBLS DNA present in T3 samples was highest at Day 14, corresponding to the active growth phase of the pathogen, while on Day 42, the quantity of DNA increased to 13.58% of Day 14 readings, indicating continuous growth in vivo. The method developed was time-sensitive and reliable for screening oil palm for response during the plant-pathogen interaction. The work examined the hypothesis that G. boninense utilizes lignin degrading enzymes (LDEs) such as laccase, lignin peroxidase (LiP) and manganese-dependent peroxidase (MnP) to breakdown oil palm lignin, causing primary cell, tissue and stem rot. Therefore, the role of laccase was investigated during the interaction in otherwise symptomless oil palm tissues. A small gene fragment (208 bp) of laccase was isolated from total DNA of G. boninense GBLS and sequencing showed it to contain 89% homology to basidiomycete laccase. GBLS reduced the total lignin content of oil palm in T3 plants (48.86% and 53.18% lower than T1 and T2, respectively). However, neither laccase nor MnP enzymes were produced in significantly higher amounts in T3 as compared to T1 and T2, indicating the need to differentiate the presence of plant and fungal laccases. Transcript abundance for GBLS laccase gene using qPCR indicated that laccase was induced during the interaction, with maximum laccase detected on Day 28. However, this did not place laccase as a virulence factor although the presence of higher amounts of laccase towards the end of the experiment corresponds to loss of lignin and plant death. Therefore, laccase and other LDEs need further investigations to be confirmed as virulence factors. This work reports a novel infection assay for G. boninense interaction with oil palm and was the first study to have investigated the role of G. boninense laccases in the devastating BSR disease.

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O uso de cogumelos para fins medicinais é muito antigo, onde o basidiomiceto Ganoderma lucidum tem posição de destaque no Japão e na tradicional medicina chinesa, sendo relatadas atividades antitumorais, antivirais, antibacteriana, dentre outras. Várias espécies com superfície lacada são confundidas com G. lucidum, entre elas Ganoderma parvulum Murrill, uma espécie tropical que pode representar potencial econômico, respeitando-se a biodiversidade local. O presente trabalho objetivou avaliar a atividade antibacteriana, concentração inibitória mínima (CIM) e a atividade citotóxica dos extratos metanólico e em acetato de etila de G. parvulum, a atividade antibacteriana e a CIM do óleo essencial, bem como fornecer dados para caracterização da droga fúngica. Para os extratos estudados, tanto a atividade antibacteriana como citotóxica foram observadas apenas para o extrato em acetato de etila. A atividade antibacteriana foi considerada moderada frente a Staphylococcus aureus, Bacillus subtilis e Micrococcus luteus. A atividade citotóxica em dose única de 50 μg/ml de extrato foi de 71,94 ± 2,04% contra a linhagem de células HT-29 e 90,02 ± 2,17% contra a linhagem de células HEp-2. O óleo essencial teve atividade antimicrobiana contra as bactérias gram-positivas (Staphylococcus aureus, Bacillus subtilis e Micrococcus luteus) e gram-negativas (Escherichia coli, Klebsiella pneumoniae e Pseudomonas aeruginosa), contudo bactérias gram-positivas foram mais susceptíveis ao óleo essencial. Quanto à prospecção micoquímica, os compostos majoritários foram representados pelos esteróides e terpenóides, apresentando também flavonóides, derivados cinâmicos e fenilpropanoglicosídeos. A composição do óleo essencial foi analisada por cromatografia gasosa acoplada a espectroscopia de massas (CG-MS), onde os principais compostos foram o ácido benzóico, o sesquiterpeno aristolona mono-oxigenado, o 2,6-bis (1,1-dimetiletil)-4-metilfenol e o N-(2-hidroxi-4-oxo-4H-quinazolina-3-il)-benzamida. A morfoanatomia foi considerada fundamental para a identificação de G. parvulum, onde características como o tamanho do basidioma, tamanho do basidiósporo, o sistema hifálico, a coloração do contexto e linhas de deposição de material resinoso, foram características essenciais para distinção de G. parvulum. O tamanho das partículas do pó indicou se tratar de um pó moderadamente espesso por simples agregação das partículas, o teor de cinzas totais foi de 1,67 ± 0,14% e a perda por dessecação foi de 13% ± 0,07, dentro dos níveis encontrados para o gênero. Na análise microscópica do pó, a observação de hifas esqueléticas e de esporos de parede dupla e truncados, característicos do gênero, foram importantes para a identificação da droga. Os dados sobre teor de cinzas, perda por dessecação, análise granulométrica e microscópica do pó são inéditos para G. parvulum, bem como as atividades biológicas dos extratos estudados e do óleo essencial.

Ganoderma lucidum spores (GLS) have attracted increasing attention for its versatile biological activities, particularly in cancer therapy. The resilient chitin bilayer of sporoderm is conventionally regarded as an obstacle in the exploitation of bioactive ingredients. Present study found that ethanol extract of broken GLS was able to inhibit cancer cells, however, water extract, especially medium extract (containing serum protein) from unprocessed GLS have also demonstrated anti-proliferative effects on cancer cells. The effectiveness of GLS extract on the inhibition of a series of human cancer cells, namely, osteosarcoma, neuroblastoma, myeloid leukaemia and breast cancer, has been compared, and DNA assays showed that the GLS extract is more efficient in inhibiting neuroblastoma but has less effect on osteosarcoma cell line. To overcome the limitations of the existing processing methods of GLS, the feasibility of sonication as a new way to break GLS has been tested. A series of processing parameters, such as sonication power and duration, have been compared to maximise the breaking efficiency. The preservation of bioactive components of GLS (e.g. polysaccharides and ganoderic acids) from sonication processing was revealed by Fourier Transform Infrared Spectroscopy (FTIR) and High Performance Liquid Chromatography (HPLC) analyses. In vitro study showed that sonication processed GLS were able to inhibit breast cancer cells, at dose and time dependent manner, particularly at low pH (6.5), favourable for cancerous cell growth. The inhibitory efficiency of sonication processed GLS on the growth of breast cancer cells was ranked the highest, compared with that of unprocessed GLS and commercially broken GLS. To preserve further the bioactive ingredients of GLS, broken GLS have been encapsulated with alginate by electrospraying (ES). The size of GLS encapsulated alginate (GLS/A) beads was found to affect the in vitro release profiles of bioactive ingredients of GLS, and can be controlled by varying the processing parameters (e.g. crosslinking time, infuse rates and applied voltage). A series of GLS/A beads with mean sizes ranging from 500 to 2500 µm have been produced by ES and the in vitro release profiles of GLS/A beads in simulated gastrointestinal mediums were found to be related to the pH, bead size and drying methods. In summary, an advanced method combining a customised sonication with ES has been developed by setting up a lab-scale production line from processing to encapsulation of GLS. This may pave the way to produce effective GLS products with desirable natural bioactive components for healthcare.

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Físicas e Matemáticas. Programa de Pós-Graduação em Química
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Ganoderma Karsten é um gênero de fungo pertencente a família

碩士
國立臺灣大學
食品科技研究所
106
Ganoderma formosanum and Ganoderma lucidium, Lingzhi, are traditional Chinese medicine and are considered as healthy food. The bioactive components in Lingzhi are (1, 3；1, 6)-β-D-glucans and ganoderic acids. This study aims to investigate that the effect of the static culture, the submerged culture and the two-stage fermentation on the production of the (1, 3；1, 6)-β-D-glucans and ganoderic acids of Ganoderma formosanum and Ganoderma lucidum. The biomass production, polysaccharide in mycelium and broth, the content and the degree of the branch of (1,3；1,6)-β-D-glucans and the structure of ganoderic acids of Ganoderma formosanum and Ganoderma lucidum were compared. In this study, the (1,3)-β-D-glucanase was used to investigate the content and the degree of the branch of (1,3；1,6)-β-D-glucans. Liquid chromatography-tandem mass spectrometry was used to analyze ganoderic acids. The structure properties of ganoderic acids were identified by accurate m/z (-2.33-2.33 ppm), MS/MS spectra and UV spectra. Standard addition method was used to analyze the concentration of ganoderic acids. Relative quantification method was used to analyze the ganoderic acids without an available commercial standard. In this research, the result shows that static culture produces more ganoderic acid than submerge culture. However, shaking culture produces more biomass and polysaccharide. Two-stage fermentation can enhance the yield of the target compound. The mycelium of Ganoderma formosanum has maximum biomass by static culture. At 10 days, Ganoderma formosanum has the maximum yield of polysaccharides in the mycelium and the broth. By shaking culture, the maximum biomass of the mycelium of Ganoderma lucidum is at 15 days and the maximum yield of the polysaccharide of Ganoderma lucidum is at 10 days but there is no significant difference in the broth. At 30 days, by static culture, the content of (1,3；1,6)-β-D-glucan of Ganoderma formosanum mycelia is more than Ganoderma lucidum. The degree of the branch of (1,3；1,6)-β-D-glucan of Ganoderma formosanum mycelia is between 0.2-0.33. Ganoderma lucidum only produces (1,3；1,6)-β-D-glucan in static culture and the degree of the branch of (1,3；1,6)-β-D-glucan is lower than 0.2-0.33. Ganoderma lucidum contains more mycelia and polysaccharides than Ganoderma formosanum. However, the percentage of (1,3；1,6)-β-D-glucan in the polysaccharides of Ganoderma formosanum is more than Ganoderma lucidum. Ganoderma lucidum and Ganoderma tsugae fruiting body were used as QC sample. 27 kinds of ganoderic acids were found in QC sample. In this study, the mycelium of Ganoderma formosanum in static culture 30 days has more ganoderic acid H than Ganoderma lucidum. The static culture and shaking culture have no significant effect on the production of Ganoderic acid H of Ganoderma lucidum.

碩士
中國醫藥大學
中國藥學暨中藥資源學系碩士班
102
Ganoderma lucidum, a popular traditional Chinese medicinal herb, has been used to treat or prevent disease for thousand years in Asia. Ganoderic acids (GAs) are one of major biologically active components and show pharmacological activities such as anti-oxidant, anti-tumor, anti-virus and anti-inflammation. However, the regulation of GAs biosynthesis is poorly understood. This study investigates the role of apoptosis signaling on GAs biosynthesis. The drugs which induced apoptosis in mammalian cells or fungi cells, such as acetic acid (AA), menadione (Vit K3), sodium chloride (NaCl), valproic acid (VPA), zinc chloride (ZnCl2), were used to treat the mycelium cell of G. lucidum. Biomass and GAs production, and cell apoptosis were evaluated. The content of total GAs was analyzed by high performance liquid chromatography (HPLC). The cell apoptosis was identified by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay (TUNEL) and nuclear morphology change. All treated drugs reduced biomass and induced GAs production. As compared with control, maximum total GAs accumulation were 1.79-fold, 2.39-fold, 1.67-fold, 2.06-fold and 1.81-fold induced by 40 mM AA, 2 mM Vit K3, 1 M NaCl, 2.5 mM VPA and 10.8 mM ZnCl2, respectively, for 4 days incubation. In addition, Vit K3 (0.5 mM to 2 mM), VPA (0.8 mM to 1.2 mM) and ZnCl2 (0.8 mM to 1 mM) induced apoptosis which were identify by positive TUNEL stain and condensed nuclear morphology. This data indicates that Vit K3, VPA and ZnCl2 induced GAs biosynthesis during cell apoptosis.

碩士
輔英科技大學
環境工程與科學系碩士班
106
Ganoderma, in a broad sense, includes the species of Ganoderma lucidum and related families, which are mainly distributed in China, the Korean Peninsula and Japan. Ganoderma lucidum, also known as triterpenoids, is the main source of bitterness in Ganoderma lucidum. It has a wide variety and is currently separated from Ganoderma lucidum by about 135 species. And it has anti-tumor, anti-HIV-1 type virus, detoxification and other activities. Extraction techniques are very diverse, such as solvent extraction, supercritical extraction and distillation. The supercritical carbon dioxide extraction method used in this study is based on the advantages of carbon dioxide being non-toxic, odorless, residue-free and chemically stable. And low temperature extraction does not cause damage to the active ingredients of the extract.In this experiment, Ganoderma lucidum extraction was carried out according to different extraction pressures and temperatures, and compared with the extracts obtained by traditional organic solvent extraction. The supercritical extraction method has a similar peak shape to the organic solvent extraction method under specific conditions. The active ingredient of Ganoderma lucidum can be extracted instead of organic solvent extraction. Finally, the optimal extraction conditions for this experiment were a pressure of 300 kg/cm2 and a temperature of 50 °C.

碩士
長庚大學
傳統中國醫學研究所
97
Ganoderma lucidum on the "shen nong ben cao jing" (The Divine Husbandmn’s Herbal Foundation Classic) is listed as the top grade of medicine during the time of Chin and Han Dynasty. But over thousand of years, the book has stated a very little about its effects. In the important remedy books such as "qian jin yi fang ", "tai ping sheng hui fang ", "sheng ji zong lu ", "yu yao yuan fang " and "pu yi fang "…etc, it has stated only five prescriptions with the Ganoderma lucidum. The quantity is very rare and makes it difficult for doctors to use it in their daily practice, but this changes to a special appearance in Taoism and local folks as an “immortal elixir” and “propitious pleasant”. In the present days, a total of 76 different types of the Ganoderma have been discovered for medical use. Due to the improving cultivating technique, and over a thousand of researches, they had made the Ganoderma lucidum to play a new role in the health medicine. The old Traditional Chinese Medicine doctors have experienced the rich variety of mushroom species in Taiwan, and re-appraisal of Ganoderma lucidum between the classic and new researches. The clinical experiences among those doctors are our enlightenments to this medical field. We had visited three old Traditional Chinese Medicine doctors, compared and investigated their opinions and thinking process in this research. Dr. Tsai has many years of clinical experiences in the use of Ganoderma lucidum, Fomitopsis pinicolat,Ganoderma applanatum, Hexagona apiaria, Ganoderma sinense, Antrodia cinnamomea, Phellinus igniarius …etc. He has widely used them on patients with liver diseases, insomnia, depression, cardiovascular diseases, renal diseases, cancer, hemorrhoids, constipation…etc. Dr. Cheng has discovered the use of Ganoderma lucidum on patients with chest tightness has a great effect on the illness. However, side effects such as edema and skin rash develop easily on patients. Due to the side effects, Dr. Cheng is no longer using it. Dr. Liu considers that the clinical effects of Ganoderma lucidum are uncertain. Therefore, he does not use any at all. Dr. Tsai only uses the one in the wild. He thinks that by cultivating, the medical effect is not so efficient. He also points out that epiphytic plants will influence the effect of Ganoderma lucidum and this can be used in improvement in cultivating technique and the reference for other doctors. These three doctors have the habits of trying the medication themselves before their use on the patients. They are not only providing a safer medication for the patients, also they are providing ones personal sensitivity to the medication. This can be used as a academic reference to its clinical use.

碩士
南台科技大學
化學工程系
91
The study utilizes Acetobacter sp. AC-01, Saccharomyces cerevisiae YC-01,YC-02 lines from kombuch and to reference kombuch basic ferments beginning sugar which amount of 10-15%. This results 15% sugar is better in bacteria growth and acetic acid yield. So fermentation of Ganoderma tsuga wine vinegar selects 15% sugar which b-grade white crystal sugar and superior white crystal sugar are basic the source of nutrition. To use two groups bacteria and two kinds basic nutrition to comparatively difference between both ferment. A group is “A”, which has three lines that Acetobacter aceti, Saccharomycaes cerevisiae and Candida sp.; another group is “Z”, which has three lines that Zygosaccharomyces rouxii, Saccharomycaes cerevisiae and Candida sp.. From this experiment results to clear apparent “A” and group Zs in b-grade white crystal sugar that are better. Total sugar maintains 15% that contains glucose or fructose makes wine vinegar fermentation. 3% glucose supports bacterium’s growth, fructose inhibitions it. Hence ultimate medium was formed with 12% b- grade white crystal sugar (or superior white crystal sugar), 3%glucose and 3% Ganoderma tsuga reside. To use the medium goes a step further probe into ferment conditions. Pure bacterium inoculated that quite plate cultivate 200ml in 250ml Erlenmeyer flask in, and fermented 4.5 liters in 5 liters fermentation tank that 50rpm, aerate trace air. The culture method that 10-fold concentrate of from sixth to fifteenth days had antimicrobial in Escherichia coli、Salmonella choleraesuis、Staphylococcus aureus、Pseudomonas aeruginosa、Proteus vulgaris、Listeria monocytogenes、 Pseudomas glumae et al. The medium of 12% b- grade white crystal sugar, 3%glucose and 3% Ganoderma tsuga reside cultivated A group As in 5 liters ferment tank. It cultivated fermentation liquid on 1, 1-diphenyl 2-picrylhdrazyl radical (DPPH·), that the highest was 83%, relativity 88.4μg/ml ascorbic acid. To compose four fermentation types that use two groups bacteria and two kinds basic nutrition that cultured 12 days in 25℃ and 800ml in laboratory bottle screw-cap of 1 liter. And the medium of 12% b- grade white crystal sugar, 3%glucose and 3% Ganoderma tsuga reside cultivated a group As 12 days in 5 liters ferment tank. Via two times from 21 and 29 persons tried drink Ganoderma tsuga wine vinegar, 20-25 years old judging workings tried drink Ganoderma tsuga wine vinegar. Accepting cultivate Ganoderma tsuga wine vinegar in quite plate and difference small. But k traditionalculture Ganoderma tsuga wine vinegar accepted than most. The medium of 12% b- grade white crystal sugar, 3%glucose and 3% Ganoderma tsuga reside cultivated a group. To cultivate in 25℃, initial stage 25℃ and a later period 28℃, in 27℃, in room temperate that was 15 days. Aceti acid content was separately 0.2, 0.5.2, 1.47, 3.40%. To cultivate in room temperate aceti acid content best high.

碩士
國立中央大學
化學工程與材料工程學系
103
Ganoderma lucidum is one of the medicinal mushrooms. The most attractive characteristic of the medicinal mushrooms is its anti-tumor effect. Ganoderma lucidum has gained wide popularity as a health food and became the most valuable mushroom in Taiwan. However, because of its host specificity, slow growth rate and rarity in nature, the fruiting bodies of Ganoderma lucidum have become expensive mushrooms in recent years. Thus, investigators have exerted their efforts to prepare this mushroom from submerged culture. The main objectives of this proposal are to develop two-stage fermentation system. In first stage, we make the Ganoderma lucidum grow in high speed for 8 days, and Saccharomyces cerevisiae was added to the fermentation broth in second stage with aerated or static process to enhance ganoderic acids production. The amount of ganoderic acids reached 171.5 mg/g DW by using static operation with 5%(v/v) of inoculation level of the Saccharomyces cerevisiae in 250 ml shake flask. A novel two stage operation to optimize triterpenoids production of a co-culture was proposed and successfully demonstrated, reaching highest ganoderic acids content obtained in this study.

Relatório de Estágio do Mestrado Integrado em Ciências Farmacêuticas apresentado à Faculdade de Farmácia
Resumo - Relatório de Estágio em Farmácia Comunitária Após a aquisição da formação teórica e prática integrada no Mestrado Integrado emCiências Farmacêuticas, o estudante realiza a unidade de “Estágio Curricular”. Sendo uma das áreas mais relevantes do domínio farmacêutico, a realização de um período de estágio em Farmácia Comunitária é obrigatória.Neste contexto, ingressei na Farmácia Coimbra, localizada no Coimbra Shopping, onde adquiri conhecimentos acerca do funcionamento interno de uma farmácia assim como experienciei o papel do farmacêutico perante a população. O presente relatório será apresentado sob a forma de análise SWOT, por forma a salientar aqueles que, segundo a minha perspetiva, foram os pontos fortes, pontos fracos, oportunidades e ameaças do estágio. Resumo - Relatório de Estágio em Indústria Farmacêutica Como parte integrante do último semestre do Mestrado Integrado em CiênciasFarmacêuticas, os estudantes têm que realizar a unidade de “Estágio Curricular”. Para além da habitual área da Farmácia Comunitária, tive a oportunidade de realizar um estágio numa outra área, mais especificamente em Indústria Farmacêutica, o que me permitiu alargar conhecimentos e explorar vertentes alternativas do domínio farmacêutico.Assim sendo, integrei a equipa da Pharmilab, uma consultora especializada na área doscosméticos, dispositivos médicos, suplementos alimentares e biocidas, onde incorporei oambiente laboratorial, com foco no Controlo da Qualidade. O presente relatório será entãouma análise desta experiência, apresentada sob a forma de análise SWOT, evidenciando ospontos fortes, pontos fracos, oportunidades e ameaças sentidos no decorrer do estágio. Resumo - Potencialidades medicinais de Ganoderma lucidum O Ganoderma lucidum é um basidiomiceto também conhecido como “cogumelo da imortalidade” em países asiáticos, onde a sua utilização remonta a épocas prévias à Era Comum. Este cogumelo tem demonstrado inúmeras potencialidades medicinais atribuídas à elevada diversidade de substâncias ativas presentes no basidiocarpo, micélio e esporos, dentrodos quais se destacam os polissacáridos e os triterpenos. A presente monografia irá abordar quatro destas potencialidades, nomeadamente, as ações imunoestimulantes, antitumorais, antidiabéticas e dislipidémicas.Quanto à estimulação do sistema imunitário salientam-se os polissacáridos, em particularos β-glucanos. Estes têm sido reportados em inúmeros ensaios in vitro e in vivo como capazes de promover a imunidade celular e humoral, assim como de estimular as célulasapresentadoras de antigénios e o sistema mononuclear fagocitário, principalmente através da sua capacidade de induzir a produção de citocinas.A ação antitumoral deste cogumelo deve-se maioritariamente à presença de triterpenos.Apesar da ação imunoestimulante dos polissacáridos ser de elevada relevância no auxílio àterapêutica do indivíduo oncológico, os triterpenos têm demonstrado uma panóplia depropriedades antitumorais, nomeadamente através das suas ações citotóxicas, citostáticas,indução da apoptose e inibição da formação de metástases e da angiogénese.Quanto à ação antidiabética, o G. lucidum tem demonstrado capacidades hipoglicemiantes ehiperinsulinémicas. Enquanto os polissacáridos têm sido reportados como reguladores deenzimas-chave do metabolismo da glucose e indutores da libertação de insulina, os triterpenos exibem capacidade de intervir nas enzimas aldose redutase e α-glicosidase. Existe ainda um proteoglicano que tem revelado propriedades inibitórias da Proteína Tirosina Fosfatase 1B.Relativamente à dislipidémia, o G. lucidum tem manifestado capacidade de promover açõesessencialmente hipocolesterolémicas. Deste modo, os polissacáridos têm sido reportadoscomo capazes de alterar a absorção intestinal do colesterol exógeno, enquanto os esteróis e derivados oxigenados do lanosterol reduzem a síntese hepática do colesterol endógeno.Neste contexto, a presente monografia procede à revisão de literatura sobre os principaisconstituintes do G. lucidum e o seu papel na imunoestimulação bem como no auxílio àterapêutica de algumas das doenças mais relevantes do presente século, por forma a enfatizar as potencialidades medicinais deste cogumelo, assim como averiguar os mecanismos de ação pelos quais atua, e a possível toxicidade e efeitos secundários inerentes à sua utilização.
Abstract - Community Pharmacy Internship Report Following the acquirement of the theoretical and practical training integrated in the Master’s Degree in Pharmaceutical Sciences, students undertake the unit “Curricular Internship”. Being one of the most relevant areas of the pharmaceutical domain, the completion of a probationary period in Community Pharmacy is mandatory.In this regard, I joined Farmácia Coimbra, located at Coimbra Shopping, where I haveknowledge about the internal functioning of a pharmacy as well as experienced thethe pharmacist’s role towards the population. The present report will be displayed under the form of a SWOT analysis, in order to underline those that, according to my perspective, were the strengths, weaknesses, opportunities and threats of the internship. Abstract - Pharmaceutical Industry Internship Report As an integrating part of the last semester of Master’s Degree in Pharmaceutical Sciences, the students have to undertake the unit “Curricular Internship”. In addition to the usual internship in Community Pharmacy, I had the opportunity to take part in another one in a distinct area, more specifically in Pharmaceutical Industry, which allowed me to expand my knowledge and explore alternative paths of the pharmaceutical domain.Therefore, I instated Pharmilab’s team, a specialized consultant in the areas of cosmetics, medical devices, dietary supplements and biocides, where I was incorporated in a laboratory environment, with a focus on Quality Control. The present report will be an assessment of this experience, presented under the form of a SWOT analysis, highlighting the strengths, weaknesses, opportunities and threats perceived throughout the course of the internship. Abstract - Medicinal Potentialities of Ganoderma lucidum Ganoderma lucidum is a basidiomycete mushroom also known as “the mushroom of immortality” in asian countries, where its utilization can be traced back to times prior to the Common Era. This mushroom has demonstrated its countless medicinal potentialities attributed to the high diversity of active substances present in the basidiocarp, mycelia and spores, within which polysaccharides and triterpenes stand out. Herein, four of these potentialities, particularly its immunostimulant, antitumor, antidiabetic and dyslipidaemic actions are approached.In terms of the immune system stimulation polyssacharides stand out, more specifically β-glucans. These have been reported in innumerous in vitro and in vivo studies as capable of promoting cellular and humoral immunity, as well as stimulating antigen presenting cells and phagocytic mononuclear system, mainly through its ability of promoting the production of cytokines.The antitumor activity of this mushroom is predominantly credited to the presence oftriterpenes. Despite the relevance of polyssacharides’ immunostimulating activity in assisting the therapeutics of the oncological patient, triterpenes have shown a panoply of antitumor properties, specifically through cytotoxicity, cytostatic actions, apoptosis induction, and metastasis formation and angiogenesis inhibition.Regarding the antidiabetic action of G. lucidum, hypoglycaemic and hyperinsulinemiccapacities have been demonstrated. While polysaccharides have been reported as modulatorsof key enzymes in the glucose metabolism pathway and as insulin release inducers, triterpenes have evidenced the ability of intervening in the aldose reductase and α-glycosidase enzymes. There is also a proteoglycan that has expressed PTP1B inhibitory properties.Lastly, concerning dyslipidaemia, G. lucidum has displayed the ability to promote essentially hypocholesterolemic actions. For that purpose, polysaccharides have been reported as capable of altering the intestinal absorption of exogenous cholesterol while sterols and oxygenated lanosterol derivatives lower the hepatic synthesis of endogenous cholesterol.Within this context, a literature review upon G. lucidum’s major components and their role in immunostimulation along with the therapeutic aid on some of the most relevant illnesses of the current century shall be carried out, in order to emphasize the medicinal potentialities of this mushroom, as well as ascertain the mechanisms of action by which it operates and the possible toxicity and secondary effects inherent in its use.

Ganoderma root rot disease has been reported world wide causing the death of affected hosts. The taxonomy of the genus Ganoderma is considered to be in disarray due to the use of basidiocarp morphological characters to differentiate the species which resulted in many synonyms, species complexes and possible misidentifications of species within the genus. The use of sexual compatibility tests and molecular techniques became powerful diagnostic tools to elucidate the taxonomy of Ganoderma species. Application of these techniques has resolved some of the taxonomic problems but the use of certain species names in the genus is still causing contention among taxonomists. The literature surrounding the taxonomy and techniques used in the taxonomy of the root rot fungus Ganoderma are considered in this thesis. It is clear that the taxonomy of Ganoderma is very difficult and it is still largely obscured by species complexes and incorrect species identifications. It is also evident that a single species concept will not aid in the identification of Ganoderma species. Rather, a combination of concepts based on morphology, mating tests and DNA sequence data should be used in elucidating the taxonomy of Ganoderma. Morphological characteristics as well as nucleotide sequence analysis of three gene regions; the internally transcribed spacer (ITS), the mitochondrial small subunit (mtSSU) and the intergenic spacer (IGS-1), were used to identify the causal agent of Ganoderma root rot of J. mimosifolia in the suburb of Brooklyn, Pretoria, South Africa. Morphological observations and DNA-based phylogenies revealed that all isolates collected from infected trees belong to a single species that reside in the G. lucidum sensu lato complex. Acacia mangium is a leguminous tree that is grown as an exotic plantation species in Indonesia. These economically important trees are threatened by Ganoderma root rot disease. This disease is considered to be the most important cause of losses in A. mangium plantations. Phylogenetic analysis of ITS sequence data showed that G. philippii is the primary agent of Ganoderma root rot in A. mangium in Sumatra, Indonesia.
Dissertation (MSc)--University of Pretoria, 2011.
Microbiology and Plant Pathology
Unrestricted

Dissertação de Mestrado Integrado em Engenharia Química apresentada à Faculdade de Ciências e Tecnologia
Os cogumelos têm vindo a ser incluídos de forma cada vez mais frequente na alimentação humana, assumindo um papel importante, quer devido às suas características nutricionais e organoléticas, quer devido às diferentes formas de consumo, despertando sensações únicas, devido ao seu aroma e sabor que colocam e despertam em cada prato. Por outro lado, são também muito consumidos, devido às suas propriedades medicinais extraordinárias, sendo cada vez mais alvo de muitos estudos realizados neste âmbito. Estudos científicos e médicos demonstram as propriedades medicinais dos compostos extraídos de cogumelos para a prevenção das doenças mais diversas, sendo cada vez mais frequentes na prevenção e tratamento do cancro (Zaidman et al., 2005; Lemieszek et al., 2013). O objetivo deste trabalho consiste na caracterização química de quatro espécies de cogumelos, Boletus edulis, Tricholoma equestre, Ganoderma lucidum e Ganoderma lingzhi e no estudo do efeito de um dos seus compostos na atividade neuronal. Neste estudo, investigou-se a ação de polissacarídeos extraídos dos cogumelos na atividade antioxidante de uma zona sináptica. Os experimentos consistem na adição de diferentes concentrações de tais extratos na região CA3 do hipocampo do cérebro no sistema sináptico de fibras musgosas usando fatias de cérebro de 400 μm de espessura.----------------------------------------------------------------------------------------------------------------------------------------------------------------
Mushrooms have been increasingly included in human food, taking on an important role, either due to their nutritional and organoleptic characteristics or due to the different forms of consumption, arousing unique sensations, due to their aroma and flavor put and wake up on each plate. On the other hand, they are also very consumed, due to their extraordinary medicinal properties, being more and more the target of many studies carried out in this scope. Scientific and medical studies demonstrate the medicinal properties of compounds extracted from mushrooms for the prevention of the most diverse diseases, being more and more frequent in the prevention and treatment of cancer (Zaidman et al., 2005; Lemieszek et al., 2013). The aim of this work is the chemical characterization of four mushroom species, Boletus edulis, Tricholoma equestre, Ganoderma lucidum and Ganoderma lingzhi, and to study the effect of one of its compounds on neuronal activity. In this study, we investigated the action of polysaccharides extracted from mushrooms on the antioxidant activity of a synaptic zone. The experiments consist of adding different concentrations of such extracts to the CA3 region of the brain hippocampus in the moss-synaptic system using 400 μm thick brain slices.-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

by Ma Suet-yee.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (leaves 119-131).
Abstracts in English and Chinese.
Acknowledgment --- p.i
Abstract --- p.ii
摘要 --- p.iv
Table of Contents --- p.vi
List of Tables --- p.x
List of Figures --- p.xii
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Ganoderma lucidum --- p.1
Chapter 1.1.1 --- History of Ganoderma lucidum --- p.1
Chapter 1.1.2 --- Classification --- p.1
Chapter 1.1.3 --- Macroscopic and microscopic structure --- p.2
Chapter 1.1.4 --- Ganoderma lucidum as a pathogen --- p.3
Chapter 1.1.5 --- Availability of tree hosts in Hong Kong --- p.4
Chapter 1.1.6 --- Medicinal effects --- p.5
Chapter 1.2 --- Study of Populations in Fungi --- p.6
Chapter 1.2.1 --- Definition of Population --- p.6
Chapter 1.2.2 --- Study of Fungal Populations --- p.7
Chapter 1.2.3 --- Techniques for Population Studies in Fungi --- p.7
Chapter 1.2.3.1 --- Somatic Incompatibility Test
Chapter 1.2.3.2 --- Isozyme Analysis
Chapter 1.2.3.3 --- Restriction Fragment Length Polymorphisms (RFLPs)
Chapter 1.2.3.4 --- Polymerase Chain Reaction (PCR) Amplification
Chapter 1.3 --- Mitochondrial DNA (mt-DNA) in Fungi --- p.14
Chapter 1.3.1 --- Inheritance in mt-DNA --- p.15
Chapter 1.3.2 --- Mitochondrial DNA in Population Studies --- p.15
Chapter 1.3.2.1 --- Mitochondrial small-subunit (mt-SSU) rDNA
Chapter 1.3.2.2 --- Cytochrome oxidase 3 (cox3)
Chapter 1.4 --- Biodiversity study on Ganoderma species --- p.19
Chapter 1.5 --- Environment Pollutants in Hong Kong --- p.20
Chapter 1.5.1 --- Air quality in Hong Kong --- p.20
Chapter 1.5.2 --- Soil quality in Hong Kong --- p.20
Chapter 1.5.3 --- Toxicity of pollutants --- p.23
Chapter 1.5.4 --- Accumulation of heavy metals by G. lucidum --- p.26
Chapter 1.6 --- Objectives of Study --- p.27
Chapter 1.7 --- Project Strategies --- p.28
Chapter 1.7.1 --- Survey on distribution and collection of Ganoderma lucidum in Hong Kong --- p.28
Chapter 1.7.2 --- Genetic divergences of G. lucidum mitochondrial genes --- p.28
Chapter 1.7.3 --- Contaminations on field collected G. lucidum --- p.29
Chapter 1.8 --- Significance of Study --- p.29
Chapter Chapter 2 --- Materials and Methods --- p.30
Chapter 2.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.30
Chapter 2.2 --- Tissue Isolation --- p.30
Chapter 2.3 --- Somatic Incompatibility Test --- p.36
Chapter 2.4 --- Molecular Identification --- p.40
Chapter 2.4.1 --- Extraction of DNA --- p.40
Chapter 2.4.2 --- Gel electrophoresis --- p.41
Chapter 2.4.3 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.41
Chapter 2.4.4 --- PCR of mt-SSU rDNA and --- p.43
Chapter 2.4.5 --- Sequencing of mt-SSU rDNA and cox3 --- p.44
Chapter 2.4.6 --- Comparison of G. lucidum complex with other Ganoderma and related species
Chapter 2. 4.7 --- Phylogenetic analyses --- p.46
Chapter 2.5 --- Investigation of pollutants in Ganoderma lucidum collected in Hong Kong --- p.46
Chapter 2.5.1 --- Metal analysis --- p.48
Chapter 2.5.1.1 --- Acid digestion
Chapter 2.5.1.2 --- Statistical analysis
Chapter 2.5.2 --- Organic pollutant analysis --- p.49
Chapter Chapter 3 --- Result --- p.52
Chapter 3.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.52
Chapter 3.1.1 --- Field observation --- p.52
Chapter 3.1.2 --- Macroscopic characteristics --- p.52
Chapter 3.1.3 --- Microscopic characteristics --- p.53
Chapter 3.2 --- Somatic Incompatibility Test --- p.56
Chapter 3.3 --- DNA fingerprints by Arbitrarily-Primed PCR --- p.57
Chapter 3.4 --- Sequencing of mt-SSU rDNA region of G. lucidum and related species --- p.60
Chapter 3.4.1 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.60
Chapter 3.4.2 --- mt-SSU rDNA region of G. lucidum and other related species --- p.61
Chapter 3.4.3 --- Phylogenetic analysis of mt-SSU rDNA region --- p.61
Chapter 3.5 --- Sequencing of cox3 region --- p.71
Chapter 3.5.1 --- Genetic variability in cox3 region of G. lucidum --- p.71
Chapter 3.5.2 --- cox3 region of G. lucidum and other related species --- p.72
Chapter 3.5.3 --- Phylogenetic analysis of cox3 region --- p.72
Chapter 3.6 --- Metal content of field G. lucidum --- p.82
Chapter 3.7 --- Organic pollutants in field collected G. lucidum --- p.90
Chapter Chapter 4 --- Discussion --- p.93
Chapter 4.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.93
Chapter 4.1.1 --- Differentiation of G. lucidum and related species in the G lucidum species complex --- p.93
Chapter 4.1.2 --- Field observation --- p.94
Chapter 4.2 --- Biodiversity of populations of G. lucidum in Hong Kong --- p.95
Chapter 4.2.1 --- Individualism of G. lucidum --- p.95
Chapter 4.2.2 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.96
Chapter 4.2.3 --- Genetic variability in cox3 region of G. lucidum --- p.98
Chapter 4.2.4 --- Lineages of G. lucidum collected in Hong Kong --- p.100
Chapter 4.2.5 --- Cryptic phylogenetic species --- p.101
Chapter 4.3 --- Contamination of field collected Ganoderma lucidum in Hong Kong --- p.106
Chapter 4.3.1 --- Metal contents in field collected G. lucidum in Hong Kong --- p.106
Chapter 4.3.1.1 --- Metal contents of G. lucidum fruiting bodies collected at each site
Chapter 4.3.1.2 --- General discussion of metals
Chapter 4.3.1.3 --- Consumption of field collected G. lucidum fruiting bodies
Chapter 4.3.2 --- Comparison of metal contents between field collected Hong Kong G. lucidum with other mushrooms collected from other places --- p.112
Chapter 4.3.3 --- Survey of organic pollutants in field collected G. lucidum in Hong Kong --- p.113
Chapter Chapter 5 --- Conclusion --- p.116
Chapter Chapter 6 --- Further investigation --- p.118
Chapter Chapter 7 --- Reference --- p.119

碩士
國立陽明大學
生物醫學資訊研究所
98
Ling-Chi (Ganoderma lucidum), a chinese herb, The recent studies have revealed that polysaccharides, triterpenes, and some microelements in Ling-Chi, that has been reported to be effective in the treatment of neoplasia, hypertension, and hepatopathy. A whole genome sequence project for Ganoderma lucidum is well-studied to understand of biosynthesis of the secondary metabolites and some microelements. Next-generation high-throughput DNA sequencing techniques are opening whole genome sequence assembled. Ling-Chi whole genome project sequenced using a combination of conventional capillary sequencer and new-generation sequencing technology, and was conducted by a collaboration of the team of the Veteran General Hospital-Taipei（VGH-Tapei）and the National Yang-Ming University（NYMU）from 2001 to present. The genome of Ling-Chi is about 45Mb in size and it contains 13 chromosomes. We predicted 14,052 genes by using gene prediction tool Genemark-ES in Ling-Chi. Moreover, these genes we used tentative unique genes (TUG) sequences applied to verify the gene structure predicted by prediction programs. The possible functions and reaction pathways of these predicted genes were annotated by using the sequence similarity to known proteins. And then, we identified some genes that might be involved in the terpenoid backbone and steroid biosynthesis by KEGG in Ling-Chi. In additional to discussing the genome assembly correctness of Ling-Chi genome, using by mate-pair short reads and TUG sequences can be applied to provide the order of contigs in genomic scaffold.

碩士
中山醫學大學
營養學研究所
93
Ganoderma lucidum played an important role in Chinese traditional medicine. Triterpenoids and polysaccharides of Ganoderma lucidum were found effective in the treatment of hypertension, chronic bronchitis, allergy, hyperglycemia and cancer. This study was focused on anti-aging effect of Ganoderma lucidum including anti-oxidation ability of plasma, erythrocyte enzyme activities and liver function protection. This was a double blind and cross over study, 39 subjects (21 males and 18 females) involved in this study. The total experimental period was thirteen months. One group of subjects took placebo in the first six months and Ganoderma lucidum in the next six months, one month between this two periods was wash out period. The other group of subjects took Ganoderma lucidum in the first six months and placebo in the next six months. Anthropometric measurements, blood collecting and abdominal ultrasound examination were performed at the initial,3rd,6th,7th,10th,13th months in the testing period. Subjects took 2.7g Ganoderma lucidum extract everyday, equivalent to 1.89g triterpenoids and 1.62g polysaccharides. Results showed that six months continuous intake of Ganoderma lucidum significantly increased total antioxidant capacity, total thiol groups, GSH, reduced TBARS and 8-OH-dG contents in plasma. The anti-oxidation enzyme activity in erythrocytes was also increased, enzymes concerned including superoxide dismutase, glucose-6-phosphate dehydrogenase, catalase and glutathione peroxidase. In addition, the intake of Ganoderma lucidum was proved effective to bring subjects with high GPT and GOT back to normal range. The abdominal ultrasound examination also showed improvement on fatty liver.

碩士
國立中興大學
生物化學研究所
89
Abstract We isolated and characterized a new compound, M8-4-1, from the methanol extract of Ganoderma amboinense according to an exclusive survey of SDBS (Integrated Spectral Data Base System for Organic Compounds). It was found to be a lanostanoid analog. The structure of M8-4-1 as indicated below, was established by NMR spectroscopy (1H-NMR, COSY, HSQC, HMBC, 13C-NMR), melting point, and chemical methods. The cytotoxic activity assay of M-8-4-1 of Ganoderma amboinense was evaluated against Huh 7 cell line. Therefore, the compound M8-4-1 is cancer cytotoxic potential. M8-4-1

碩士
中國醫藥大學
中國藥學研究所碩士班
97
Ganoderma lucidum produces polysaccharide and triterpenoids with biological activity such as anti-tumor, anti-hypertension, anti-hyperglycemia and displays strong immunomodulatory activity. However, the production of basidiocarp of G. l. takes at least 3 to 5 months. The purpose of this study is to evaluate the factors affecting mycelial biomass, polysaccharide and ganoderic acid production in different cultural condition. G. l. BCRC 36111 was cultured for 4 weeks on A and B medium, B medium with shaking or non- shaking.The mycelial biomass and ganoderic acid Me were 0.19( g/ plate )and 1.55(mg /100mg biomass) in A medium respectively, which were greater than B medium without shaking ( 0.11 g/plate and 0.20mg /100mg biomass) and B medium with shaking(0.09 g/plate and 0.48mg /100mg biomass).The production of biomass and ganoderic acid Me of BCRC 36674 was also increased by cultivation in A medium than in B medium with shaking or non- shaking. Lower water potential of medium did not increase ganoderic acid Me production in BCRC 36111. Comparing the effect of different nutritional medium(A and SGCM),the liquid SGCM medium resulted in more biomass production but much lower ganoderic acid Me production.Production of ganoderic acids was similar in A and B medium.However, G. l. BCRC 36111 strain produced more ganoderic acid than BCRC 36674.When the Intracellular polysaccharide was evaluated, the production of polysaccharide was enhanced by cultivation in A medium than in B medium.In addition, BCRC 36674 strain produced more intracellular polysaccharide than strain BCRC 36111.Our data conclude that A medium provides better production of biomass and ganoderic acid Me.

碩士
嘉南藥理科技大學
生物科技系暨研究所
91
The purpose of this study is to isolate and purify the tyrosinase inhibitor from Ganoderma lucidum. Tyrosinase is a key enzyme in melanogenesis, therefore tyrosinase inhibitor can regulate the enzyme activity of tyrosinase and prevent the undesirable effect of skin as well as browning reaction of food. The tyrosinase inhibitor from Ganoderma lucidum was extracted by various extraction media, and that extracted by 25mM sodium phosphate buffer pH6.8 showed the highest extraction efficiency, followed by methanol?z n-butanol?z n- hexane and ethyl acetate . The inhibitor was then stored at -18℃ and 4℃, the relative inhibition activity of tyrosinase were 98% and 88% after 6 month storage, respectively. The tyrosinase inhibitor from Ganoderma lucidum was further purified with　Amberlite XAD-7?z Sephadex G-25 and Superdex Peptide column chromatography, and the recovery was 83%?z 50% and 15%, respectively; the inhibitory concentration caused 50% inhibition (IC50) was 5.8?z 3.19 and 0. 15mg/ml, respectively. The inhibition activity of tyrosinase was 47% when preincubated with the inhibitor for 0 minute while 100% for 18 hour, indicating a slow binding behavior. The optimal temperature of the inhibition activity of tyrosinase treated with the inhibitor was 45℃-55℃; optimal pH 5. 0. The inhibitor was stable at pH 4.0-9.0. In Kinetic study, the inhibitor showed mixed types and the inhibitory constant (Ki) 1.1mg/ml using L-DOPA as substrate, while mixed types and Ki 0.3mg/ml using L-tyrosine as substrate. Moreover, molecular mass of the partially purified tyrosinase inhibitor from Ganoderma lucidum was 0.3kDa examined by Superdex Peptide column gel filtration. The tyrosinase inhibitor from Ganoderma lucidum may contain sugar residue or amino group, according to the positive reaction with Fehling reagent and ninhydrin.

碩士
台北醫學院
醫學研究所
84
AbstractIn order to completely utilize the polysaccharide isolated from the fruiting body of ''Ling Zhi'', this study used the most common cultured species in Taiwan, the mushroom Songshan lingzhi (Ganoderma tsugae,Murrill), as its material. We extracted three polysaccharides by different processes such as ultrafiltration, semimembrane dialysis, and lyophilization. The purpose of the study is to compare the chemical and antitumor properties of the polysaccharide produced by 1. F-I (water soluble polysaccharide), 2. F-II (alkali soluble polysaccharide), and 3. F-III (the deacetylated cell wall skeleton). The chemical analysis included total sugar assay, sugar component, ratio of the sugar component, and molecular weight. The methods involved in the analysis are : TLC, HPLC, Gel-filtratiion, and Acid hydrolysis. Antitumor activities of these polysaccharides were assayed by challenging Sarcoma 180 tumor cells to Balb/c mice and prior to which the polysaccharides were administrated into cadal vein, intravenously. The pulmonary nodules of S-180 induced by metastasis were counted by flow cytometer and naked eyes. In addition, the lymphocyte subsets in peripheral blood were measured in respect to the ratio of T, T4, T8, B, and Macrophage. In this experiment, we tried to utilize the residue of Ling Zhi which is used to be dumped. In the whole process, two additional polysaccharides were produced in large quantity and we expect these polysaccharides functioned as well as that from water soluble one. These results indicate that F-I and F-II had no much difference in their components. They were the polymers of glucose and N-acetyl-D-glucosamine (or glucosamine). The major component of F-III is glucosamine (60%). In addition, to the G2+M(%) phase of murine S-180 sarcoma cells, the inhibitory effect is F-III (38.28%) > F-I (64.30%) > F-II (79.84%) > control (100%).

碩士
台北醫學院
藥學研究所
84
Development of pharmaceutical excipient from natural plants is a continuing process. These new excipients were used widely in various dosage forms, especially tablet dosage form. The fiber obtained from the fruiting body of Ganoderma, which is a traditional Chinese medicine, belongs to fungal polysaccharide. There has no report about it's toxicity till now. In this study, the possible uses of Ganoderma fiber as tabletting excipient were examined. In the first part, the effect of treated conditions on the physical characteristics of Ganoderma powder was compared. Ganoderma fiber was treatedfor 3 hours under three different conditions: in distilled water at ambient temperature (A), in hot distilled water at 85℃ (B) and 1N NaOH solution at 85℃. After processing, the fiberwas bleached with Clorox (sodiumhypochlorite) and then tray-dried in a hot air oven. All the drying products, named modifed Ganoderma, possess good flowability. But only the powder obtained from condition C shows better compactibility. These modified Ganoderma powders were physically mixed with acetaminophen(ACT) in three different ratios (1/1, 1/2 and 1/3) and then compressed into tablets with five compaction forces(0.5, 1.0, 1.5, 2.0 and 3.0 ton). All the tablets have high crushing strength,low friability and instant disintegration properties. The crushing strength ofall tablets increases as compaction force increases, but the rate of drug released was not affected by compaction force. Most shows that more than 80 %of drug was released at 5 minutes, and dissolution was completed within approximate 10 minutes. In the second part, the physical characteristics of codried powder ofmodified Ganoderma (treated with alkaline solution) and microcrystalline cellulose (MCC) at different ratios (1:1, 1:3, 1:5 and 1:10) and the resulting tablets were examined and compared. No matter what ratio was between modified Ganoderma and MCC the flowability of powder was demonstrated to be fair. The disintegration of tablet was not affected by compaction force, but increases with the increasing content of modified Ganoderma . The crushing strength ofall tablets is higher than 10 Kp except for the tablets prepared with an 1:1 ratio of modified Ganoderma to MCC. In the third part, the tabletting characteristics of ACT by codrying with amixture of modified Ganoderma and MCC was tested. The crushing strength of most ACT tablets made with codried powder containing modified Ganoderma (treatedwith alkaline solution) and MCC increases as compaction force increases. Butthere shows rapiddecline when compressed tablets at a force higher than 2 tons. The drug releaserate from the tablets compressed at 0.5 ton increases as modified Ganoderma content increases. But when compaction force was over 1 tonthe release rates were not influenced by compaction force as well as by the content of modified Ganoderma. However, the dissolution of ACT from these tablets compressed at 1 ton could be sustained to longer than 24 hours. The extent of drug release was shownto be increased with increasing amount of modified Ganoderma in the codried mixture. The addition of disintegrants could further accelerate the drug released from tablet. It was also dependent on the amount and kind of disintrgrant added. The disintegration rate was in the following order : Primojel > Crospovidone > Starch 1500.

碩士
國立臺灣大學
生化科技學系
106
Ganoderma formosanum is a unique species of Ganoderma isolated in Taiwan. Our previous studies showed that PS-F2, an extracellular polysaccharide fraction purified from the submerged culture of G. formosanum, could active macrophages by Toll-like receptor (TLR)-4, complement receptor (CR) 3 and Dectin-1, and serve as a Th1 adjuvant in vivo. PS-F2 also exhibits antitumor activity when given intraperitoneally or orally to mice. In this study, we investigated whether PS-F2 exerts the antitumor activity via modulating the polarization of M2-like tumor-associated macrophages (TAMs). Our data showed that in both M0 and IL-4-polarized M2 bone marrow derived macrophages (BMDMs), PS-F2 stimulated the production of proinflammatory cytokines TNF-α and IL-6, and the surface expression of CD86, CD40. In addition, PS-F2 stimulated the expression of M1 macrophage signature genes iNOS, IL-12, IL-6, IL-1β, IFN-β and TNF-α, while downregulating the expression of M2 macrophage-related genes arginase1, TGF-β, and Ym-1. Similar results were obtained when primary TAMs isolated from C26 tumor-bearing mice were treated with PS-F2. Furthermore, when M2 macrophages were treated with PS-F2, their suppressive effect on T cell proliferation was alleviated. Taken together, our data demonstrate that PS-F2 stimulation can repolarize protumor M2-like TAMs into antitumor M1 macrophages, resulting in the suppression of tumor growth.

碩士
靜宜大學
應用化學研究所
98
Liposomes are spheres made of lipid bilayers, enclosing aqueous volumes. The advantage of liposomal delivery is that separates the functions required for drug delivery. During the 1990s, a number of lipsome-based drugs reached the market in the U.S. and Europe and there has been an explosion of new procedures and techniques in the field. Ganoderma lucidum is used as an edible medicinal material in Chinese medicine. In recent years, many scientific researches have shown G. lucidum has a wide variety of functions. The research focuses on combining liposome with G. lucidum by using thin-film method in order to prepare the liposomes encapsulated G. lucidum extract, and to evaluate its physical stability concerning the particle size and encapsulation efficiency. Based on this study, we found that the optimal drug/lipid ratio still remains high encapsulation efficiency above 61%, and particle size is less than 15.24 μm after three months storage at 4℃.

碩士
國立臺灣師範大學
化學學系
82
Ganoderma comphoratum Zang et Su, SP. nov., growing rarely on the inner cavity wall of Cinnamomum comphora (L) Prcsl. in Taiwan, is a new genus of Ganoderma species. The methanolic extract of the fruiting body of the ganoderma comphoratum is separated by column chromatography, normal phase high performance liquid chromatography. As a result, thirteen compounds were isolated and identified by the spectral analysis. These Compounds are: 　　＊Compound A：4α-Methylergost-8, 24 (28)-dien-3, 11-dion-26-oic acid. 　　　Compound B：4α-Methylergost-8, 24 (28)-dien-3, 7, 11-trion -26-oic acid. 　　＊Compound C：7β-Hydroxy-4a-methylergost-8, 24(28)-dien-3, 11-dion-26-oic acid. 　　　Compound D：7β-Hydroxy-4a-methylergost-8, 24(28)-dien-3, 7, 11-trion-26-oic acid. 　　＊Compound E：4α-Methylergost-8, 14, 24 (28)-trien-3, 11-dion-26-oic acid. 　　＊Compound F：7β-Hydroxy-4α-methylergost-8, 14, 24(28)-trien-3, 11-dion-26-oic acid. 　　＊Compound G：Methyl-3α, 2α-dihydroxy-4α-methylergost-8, 24(28)-dien-7, 11-dion-26-oate. 　　＊Compound H：2, 2'', 5, 5'' -tetramethoxy-3, 4, 3'', 4''-di-methylenedioxy-6, 6''-dimethyl biphenyl. 　　　Compound I：3β-Hydroxy-24-methylenelanost-7, 9(11)-dien-21-oic acid. 　　　Compound J：3β, 15α-Dihydroxy-24-methylenelanost-7, 9(11)-dien-21-oic acid. 　　　Compound K：24-Methylenedihydrolanosterol. 　　　Compound L：9(E)-Octadecenoic acid. 　　　Compound M：9, 12(E, E)-Octadecadienoic acid. 　　There were six new compounds (＊ symbol) which haven''t reported on any paper.

by Cheung Ka Wan.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 121-128).
Abstracts in English and Chinese.
Chapter Chapter 1 - --- Introduction --- p.1
Chapter 1.1 --- Plant Pathogens --- p.1
Chapter 1.1.1 --- Virus --- p.2
Chapter 1.1.2 --- Viroids --- p.2
Chapter 1.1.3 --- Bacteria --- p.3
Chapter 1.1.4 --- Fungi --- p.3
Chapter 1.1.5 --- Mycoplasma like organisms --- p.3
Chapter 1.1.6 --- Nematodes --- p.4
Chapter 1.1.7 --- Insects --- p.4
Chapter 1.1.8 --- Mammals --- p.4
Chapter 1.2 --- Pathogenicity --- p.5
Chapter 1.3 --- Disease Development --- p.6
Chapter 1.3.1 --- Primary infection --- p.6
Chapter 1.3.2 --- Penetration to host --- p.6
Chapter 1.3.2.1 --- Entry through wounds ^
Chapter 1.3.2.2 --- Entry through natural openings --- p.8
Chapter 1.3.2.3 --- Direct penetration
Chapter 1.3.3 --- Colonization of pathogen --- p.9
Chapter 1.3.4 --- Mechanisms of attack --- p.9
Chapter 1.3.5 --- Symptom expression --- p.11
Chapter 1.3.6 --- Spread of disease --- p.11
Chapter 1.4 --- Detection of Pathogen --- p.12
Chapter 1.4.1 --- Traditional diagnostic methods --- p.12
Chapter 1.4.2 --- Molecular diagnostic methods --- p.13
Chapter 1.4.3 --- Advantages of molecular diagnostic tools over traditional detection methods --- p.14
Chapter 1.4.4 --- Sensitivity of molecular diagnostic tools --- p.14
Chapter 1.4.5 --- Polymerase chain reaction (PCR) --- p.15
Chapter 1.4.5.1 --- Mechanism of PCR --- p.16
Chapter 1.4.5.2 --- Application of PCR --- p.17
Chapter 1.4.6 --- Designation of specific primers in pathogen detection --- p.17
Chapter 1.4.6.1 --- Nuclear ribosomal DNA genes --- p.18
Chapter 1.4.6.2 --- Sequencing of ITS regions of rDNA --- p.19
Chapter 1.5 --- Ganoderma lucidum Complex --- p.19
Chapter 1.5.1 --- History of Ganoderma lucidum complex --- p.19
Chapter 1.5.2 --- Classification --- p.20
Chapter 1.5.3 --- Macroscopic and microscopic structure --- p.21
Chapter 1.5.4 --- Species identification in G. lucidum complex --- p.22
Chapter 1.5.5 --- Ganoderma species in Hong Kong --- p.23
Chapter 1.5.6 --- Act as pathogen --- p.25
Chapter 1.5.7 --- Availability of tree hosts in Hong Kong --- p.25
Chapter 1.5.7.1 --- Acacia confusa --- p.26
Chapter 1.5.7.2 --- Listea cubeba --- p.26
Chapter 1.5.7.3 --- Leucaena leucocephala --- p.27
Chapter 1.5.8 --- Disease control for Ganoderma lucidum --- p.27
Chapter 1.6 --- Aims of Study --- p.29
Chapter 1.7 --- Significance of the Study --- p.29
Chapter 1.8 --- Project Strategies --- p.30
Chapter 1.8.1 --- Survey on Ganoderma lucidum complex in Hong Kong --- p.30
Chapter 1.8.2 --- Artificial infection --- p.30
Chapter 1.8.3 --- Detection of pathogen --- p.30
Chapter Chapter 2 - --- Materials and Methods --- p.31
Chapter 2.1 --- Collection of Ganoderma lucidum Species Complex in Hong Kong --- p.31
Chapter 2.2 --- Tissue Isolation --- p.31
Chapter 2.3 --- Molecular Identification --- p.45
Chapter 2.3.1 --- Extraction of DNA --- p.45
Chapter 2.3.2 --- Gel Electrophoresis --- p.45
Chapter 2.3.3 --- Sequencing of ITS 1 and ITS2 --- p.46
Chapter 2.3.4 --- Comparison of G. lucidum complex with other Ganoderma and related species --- p.48
Chapter 2.3.5 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.49
Chapter 2.4. --- Mating Compatibility for Species Delimitation --- p.49
Chapter 2.4.1 --- Protoplast isolation --- p.49
Chapter 2.4.2 --- Mon-Mon mating --- p.50
Chapter 2.4.3 --- Di-Mon mating --- p.50
Chapter 2.5 --- Preparation of Samples for Scanning Electron Microscope (SEM) --- p.51
Chapter 2.6 --- Cytological Studies of Basidiocarps of G. lucidum --- p.52
Chapter 2.7 --- Pathogenicity Study --- p.53
Chapter 2.7.1 --- Growth and spread of G. lucidum in soil --- p.53
Chapter 2.7.2 --- Colonization of G. lucidum on different organs of plants --- p.53
Chapter 2.7.2.1 --- Determination of dry weight loss --- p.53
Chapter 2.7.2.2 --- Chitin assay --- p.54
Chapter 2.7.3 --- Artificial infection to tree seedlings --- p.54
Chapter 2.7.3.1 --- Artificial infection of vegetative mycelia --- p.54
Chapter 2.7.3.2 --- Artificial infection with basidiospores --- p.56
Chapter Chapter 3 - --- Results --- p.57
Chapter 3.1 --- Collection of Ganoderma lucidum Complex in Hong Kong --- p.57
Chapter 3.1.1 --- Macroscopic characteristics --- p.57
Chapter 3.1.2 --- Microscopic characteristics --- p.57
Chapter 3.1.3 --- G. lucidum under scanning electron microscopy --- p.59
Chapter 3.2 --- Field Observation --- p.62
Chapter 3.3 --- Sequencing of ITS Region of G. lucidum Complex and Related Species --- p.64
Chapter 3.3.1 --- ITS 1 Region of G. lucidum --- p.66
Chapter 3.3.2 --- ITS 2 Region of G. lucidum --- p.68
Chapter 3.3.3 --- Relationship between Ganoderma and related species --- p.71
Chapter 3.4 --- Species Delimitation of G. lucidum --- p.74
Chapter 3.4.1 --- Arbitrarily-Primed PCR --- p.74
Chapter 3.4.2 --- Di-Mon mating --- p.77
Chapter 3.5 --- Pathogenicity of G. lucidum --- p.81
Chapter 3.5.1 --- Growth and spread in soil --- p.81
Chapter 3.5.2. --- Preference in colonization on different organs of plants --- p.81
Chapter 3.5.3 --- Artificial infection --- p.87
Chapter Chapter 4 - --- Discussion --- p.98
Chapter 4.1 --- Ganoderma lucidum Complex in Hong Kong --- p.98
Chapter 4.1.1 --- Macroscopic and microscopic characteristics of G. lucidum complex and related species --- p.98
Chapter 4.1.2 --- Cytological studies --- p.99
Chapter 4.1.3 --- Field observation --- p.100
Chapter 4.1.4 --- Sequences of ITS regions of G. lucidum complex and related species --- p.101
Chapter 4.1.5 --- Species identification within G. lucidum --- p.104
Chapter 4.2 --- Pathogenicity Test for G. lucidum --- p.108
Chapter 4.2.1 --- Growth and spread of G. lucidum --- p.108
Chapter 4.2.2 --- Colonization of G. lucidum on plants --- p.110
Chapter 4.2.3 --- Artificial infection by G. lucidum --- p.111
Chapter 4.3 --- Further Investigation --- p.116
Chapter Chapter 5 - --- Summary --- p.118
Chapter Chapter 6- --- Conclusion --- p.120
References --- p.121

碩士
國立臺灣大學
食品科技研究所
94
Ganoderma lucidum is one of the most well known Chinese medicine in ancient times and is considered as the most valuable plant for health and longevity promotion. In this study, we investigated the effects of Ganoderma lucidum liquid fermentation broth on immunomodulatory activities in vitro and in vivo. After an in vitro experiment, we selected the most active fermentation product for experiments on non-specific immune response in BALB/c mice. In addition, we observed the MNC proliferation effect and performed the proteomic analysis of the protein expression by human mononuclear cells (MNC) treated with ethanol precipitated Ganoderma lucidum polysaccharide. Results show that Ganoderma lucidum fermentation product GL-93-12-21 stimulates human mononuclear cells to secrete cytokines and inhibit the human myelocytic lymphoma U937 cell growth. Moreover, the ethanol precipitated polysaccharide stimulats NO and TNF-α production in RAW 264.7 macrophage, and the polysaccharide characteristics are similar with bioactive polysaccharides that have been reported. In vivo studies showed that oral administration of Ganoderma lucidum fermentation product GL-93-12-21 on BALB/c mice significantly (P＜0.05) increases the secretion of IgG, the proliferation of T- and B-lymphocyte, the level of TNF-α、IL-2 and IFN-γ production, the phagocytosis effect of monocytes and the cytotoxicity of nature killer cells. Results also show that Ganoderma lucidum polysaccharide stimulates the proliferation of human mononuclear cells and changes the protein expression. There are 19 protein spots (1 down-regulated and 18 up-regulated) differentially expressed more than 1.5-fold, which was identified by quantitative image analysis and matrix assisted laser desorption ionization-quadrupole-time of flight (MALDI-Q-TOF) mass spectrometry. The Swiss-Prot database search indicated that increased expression of cytoskeleton and ribonucleoprotein is associated with the immune cell activation and proliferation. Furthermore, the increase of annexin A6 and chaperon protein expression, may play a part in activating of B lymphocyte and assembling of B cell receptor.

碩士
國立陽明大學
生物資訊研究所
94
Ling-Chi (Ganoderma lucidum) is one of the traditional Chinese herbs. The recent studies have revealed that polysaccharides, triterpenes, and some microelements in Ling-Chi have several interesting effects, such as anti-tumor, hypoglycemic, hypocholesterol, and immuno-stimulation. The genome and expressed sequence tag (EST) analyses are useful in identifying the genes involved in the biosynthesis of polysaccharides or triterpenes. Ling-Chi genome project have sequenced the draft sequence by shotgun method. In order to discover the expressed genes, the EST project has completed the sequencing of 47,122 EST sequences in 2005. The redundancy of the EST sequences was removed by two steps. The EST sequences were assembled and then be compared with the known protein sequences of other species. Those sequences that match the same protein were merged and got 9,697 tentative unique genes (TUG-1). The possible functions and reaction pathways of these TUG’s were annotated by using the sequence similarity to known proteins. On the basis of literature survey and sequence analysis, I identified 46 genes that might be involved in the polysaccharide biosynthesis in Ling-Chi. Moreover, 20 genes among the 46 genes, which are very similar to the known genes in the database, were annotated further. The gene structure, the predicted protein sequences, the putative protein domains of these genes were summarized for functional studies. In additional to annotating the function of Ling-Chi genes, EST sequences can also be applied to verify the gene structure predicted by programs, to estimate the gene numbers, to provide the order of genomic contigs, and to find the putative paralogous genes of Ling-Chi. Selected examples are discussed to demonstrate the feasibility of such approaches.

碩士
東吳大學
微生物學系
95
White-rot fungi is the most common degrader in lignin degradation. They produce three major extra cellular lignolytic enzymes, laccase, manganese peroxidase and lignin peroxidase. One of genus responsible for white-rot in wood is Ganoderma. This genus has been extensively studied due to some of the medicinal properties. However, we will focus on the conception of the potentially ligninolytic application. There are four isolated Ganoderma sp. were obtained for the degradation of polycyclic aromatic hydrocarbons (PAHs), containing phenanthrene and pyrene. After incubating 12 days, CCRC36021 and M-216 had better degradation abilities for PAHs. CCRC36021 was able to degrade 88.3% of phenanthrene, and 87.0% of pyrene. M-216 was able to degrade the 76.4% of phenanthrene, and 71.5% of pyrene. CCRC 36021’s degradation ability for PAHs was enhanced when the presence of Cu2+ or citric acid. Phenanthrene and pyrene were mostly degraded at 90.3 % and 83.4 %, respectively. The production of laccase(452 U/L) without the treatment of media was further enhanced to 800 and 636 U/L with the presence of Cu2+ or citric acid, respectively. Furthermore, M-216’s degradation ability for PAHs was enhanced when the presence of guaiacol. Phenanthrene and pyrene were mostly degraded at 73.8 %, 71.8%, respectively. The production of laccase (154 U/L) without the treatment of media was further enhanced to 504 U/L when the presence of guaiacol. On the other hand, both strains’ degradation abilities were reduced by adding oxalic acid. The optimal pH value for PAHs degradation was pH 4 for CCRC 36021. The maximum laccase and manganese peroxidase activities were 814 and 96 U/L. After incubating 12 days, we applied the supernatants from CCRC36021 and M-216 to treat with PAHs. We found the supernatants from both strains were able to degrade 80% and 68% of phenanthrene, respectively. Also, the supernatants degraded 68% and 61% of pyrene, respectively. In addition, after having the supernatant go through molecular sieve, we found the molecular weight of laccase was above 50 kDa. The optimal condition for laccase production from CCRC36021 is under pH 4 and 30 ℃. The enzymatic activity remained only 20% and 0% of its initial activity after thermal treatment at 70℃ for 60 minutes and 240 minutes. At pH 5, the enzyme was completely destroyed. The optimal condition for manganese peroxidase production from CCRC36021 is under pH 4 and 37℃. It remained only 30 % and 10% of its initial activity after thermal treatment at 70℃ for 60 minutes and 240 minutes. At pH 5, the enzyme was completely inactivated. Addition of 1 mM CaCl2 efficiently prevented the thermal inactivation of manganese peroxidase from CCRC36021. At 70℃, 1 mM CaCl2 elevated 10% of the enzymatic activity of manganese peroxidase. In our experiment, laccase and manganese peroxidase were both sensitive to high temperature. To sum up, CCRC36021 was able to degrade PAHs, and the highest degradation ability was achieved by using gallic acid、citric acid and Cu2+ along with low nitrogen medium (pH 4).