Thesis (M. Med. Sc.)--University of Hong Kong, 2010. / Includes bibliographical references (p. 55-61).

Sun Baizhong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 75-79). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ABSTRACT (IN CHINESE) --- p.ii / TABLE OF CONTENTS --- p.iii / LIST OF FIGURES --- p.v / LIST OF TABLES --- p.vii / ABBREVIATIONS --- p.vi / ACKNOWLEDGEMENT --- p.ix / DECLARATION --- p.x / Chapter CHAPTER ONE --- INTRODUCTION / Chapter 1.1 --- Polysaccharides --- p.1 / Chapter 1.2 --- Pharmacological importance of polysaccharides --- p.2 / Chapter 1.3 --- Polysaccharides from Ganoderma Lucidum --- p.3 / Chapter 1.4 --- Characterization of polysaccharides --- p.3 / Chapter 1.5 --- Matrix-assisted laser desorption / ionization --- p.5 / Chapter 1.6 --- Matrix-assisted laser desorption / ionization of polysaccharides --- p.8 / Chapter 1.7 --- Outline of the project --- p.10 / Chapter CHAPTER TWO --- INSTRUMENTATION AND EXPERIMENTAL / Chapter 2.1 --- Time-of-flight mass spectrometry --- p.12 / Chapter 2.1.1 --- Instrumentation --- p.15 / Chapter 2.1.1.1 --- Laser system --- p.16 / Chapter 2.1.1.2 --- Ion source --- p.16 / Chapter 2.1.1.3 --- Reflector --- p.19 / Chapter 2.1.1.4 --- Detector --- p.19 / Chapter 2.1.1.5 --- Data acquisition and manipulation --- p.20 / Chapter 2.2 --- Ultraviolet-visible spectrometer --- p.21 / Chapter CHAPTER THREE --- OPTIMIZATION OF MALDI CONDITIONS FOR POLYSACCHARIDE ANALYSIS USING DMSO AS SOLVENT / Chapter 3.1 --- Introduction --- p.22 / Chapter 3.2 --- Experimental --- p.25 / Chapter 3.2.1 --- Selection of matrix materials --- p.26 / Chapter 3.2.2 --- Selection of co-matrix materials --- p.26 / Chapter 3.2.3 --- Ratios of matrix-to-analyte --- p.26 / Chapter 3.2.4 --- Effect of sample drying temperature --- p.27 / Chapter 3.2.5 --- Sample loading methods --- p.27 / Chapter 3.3 --- Results and discussion --- p.27 / Chapter 3.3.1 --- Selection of matrix materials --- p.28 / Chapter 3.3.2 --- Selection of co-matrix materials --- p.30 / Chapter 3.3.3 --- Effect of matrix-to-co-matrix ratio --- p.32 / Chapter 3.3.4 --- Effect of sample drying temperature --- p.37 / Chapter 3.3.5 --- Effect of matrix-to-analyte ratio --- p.39 / Chapter 3.3.6 --- Evaluation of sample preparation protocols --- p.43 / Chapter 3.4 --- Conclusion --- p.46 / Chapter CHAPTER FOUR --- MOLECULAR AND MOLECULAR WEIGHT DISTRIBUTION OF WATER-INSOLUBLE POLYSACCHARIDES FROM GANODERMA LUCIDUM / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Experimental --- p.48 / Chapter 4.2.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.48 / Chapter 4.2.2 --- Gel permeation chromatography of water-insoluble polysaccharides --- p.49 / Chapter 4.2.3 --- Phenol sulfuric acid assay --- p.51 / Chapter 4.2.4 --- MALDI-TOF analysis --- p.51 / Chapter 4.2.5 --- Bradford assay --- p.52 / Chapter 4.3 --- Results and discussion --- p.52 / Chapter 4.3.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.52 / Chapter 4.3.2 --- Fractionation of water-insoluble polysaccharides using gel permeation chromatography (GPC) --- p.54 / Chapter 4.3.3 --- MALDI-TOF analysis of fractionated polysaccharides --- p.57 / Chapter 4.3.4 --- Carbohydrate content analysis of fractionated polysaccharides --- p.62 / Chapter 4.3.5 --- MW and MWD of water-insoluble polysaccharides extracted from Ganoderma Lucidum --- p.69 / Chapter 4.4 --- Conclusion --- p.71 / Chapter CHAPTER FIVE --- CONCLUDING REMARKS / Chapter 5.1 --- Conclusion --- p.73 / REFERENCES --- p.76 / APPENDIX --- p.81 / Appendix A Reaction scheme of carbohydrate with phenol in phenol-sulfuric acid assay --- p.81

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

by Fu Sai-chuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 102-111). / Acknowledgements / List of Abbreviation / Abstract / Prologue --- p.1 / Chapter / Chapter 1. --- Introduction --- p.2 / Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4 / Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6 / Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6 / Chapter 1.3.1 --- The triterpenoids --- p.7 / Chapter 1.3.2 --- Matrix polysaccharides --- p.8 / Chapter 1.4 --- Medicinal properties and biological activities --- p.9 / Chapter 1.4.1 --- Medicinal properties --- p.9 / Chapter 1.4.2 --- Biological activities --- p.10 / Chapter 1.4.2.1 --- Anti-tumour activity --- p.10 / Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11 / Chapter 1.4.2.3 --- Effects on nervous system --- p.11 / Chapter 1.4.2.4 --- Effects on respiratory system --- p.12 / Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12 / Chapter 1.4.2.6 --- Effects on immune system --- p.14 / Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14 / Chapter 1.4.3 --- Summary --- p.15 / Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15 / Chapter 2. --- Materials and Methods --- p.20 / Chapter 2.1 --- Cell culture reagents / Chapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20 / Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20 / Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20 / Chapter 2.1.4 --- Mitogens and test samples --- p.21 / Chapter 2.1.5 --- Trypan blue exclusion test --- p.21 / Chapter 2.1.6 --- Liquid scintillation counting --- p.21 / Chapter 2.2 --- Animal care --- p.22 / Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22 / Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23 / Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23 / Chapter 2.4.2 --- Preparation of liver homogenates --- p.23 / Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25 / Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25 / Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25 / Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26 / Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26 / Chapter 2.5 --- Preparation of murine hepatocytes --- p.27 / Chapter 2.5.1 --- Liver perfusion --- p.27 / Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29 / Chapter 2.5.3 --- Incubation protocol --- p.29 / Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30 / Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31 / Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31 / Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31 / Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32 / Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34 / Chapter 2.7 --- Chromatographic methods --- p.34 / Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34 / Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35 / Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35 / Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36 / Chapter 2.8 --- Measurement of in vivo antibody production --- p.39 / Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39 / Chapter 2.8.2 --- Direct plaque assay --- p.39 / Chapter 2.9 --- Biochemical analysis --- p.39 / Chapter 2.9.1 --- Determination of protein content --- p.40 / Chapter 2.9.2 --- Determination of hexose content --- p.40 / Chapter 2.9.3 --- Determination of uronic acid content --- p.40 / Chapter 2.9.4 --- Determination of sulphate content --- p.41 / Chapter 2.9.5 --- Determination of hexosamine content --- p.42 / Chapter 2.10 --- Statistical analysis --- p.42 / Chapter 3. --- Results --- p.44 / Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44 / Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47 / Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47 / Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47 / Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51 / Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51 / Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51 / Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55 / Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55 / Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55 / Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58 / Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65 / Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65 / Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69 / Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69 / Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72 / Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76 / Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76 / Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76 / Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80 / Chapter 4. --- Discussions --- p.83 / Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83 / Chapter 4.2 --- The significance of enhancement of GSH status --- p.89 / Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95 / Conclusions --- p.99 / Appendix --- p.100 / Reference --- p.102

Li Pik Ha Ivy. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 83-87). / Abstracts in English and Chinese. / TABLE OF CONTENTS --- p.i / LIST OF FIGURES --- p.v / LIST OF TABLES --- p.vii / ABSTRACT --- p.viii / ACKNOWLEDGEMENT --- p.x / DECLARATION --- p.xi / ABBREVIATIONS --- p.xii / Chapter Chapter one --- Introduction / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Polysaccharides isolated from Ganoderma Lucidum --- p.4 / Chapter 1.3 --- Conventional methods for molecular weight (MW) determination of polysaccharides --- p.6 / Chapter 1.3.1 --- Osmometry --- p.7 / Chapter 1.3.2 --- Light Scattering --- p.8 / Chapter 1.3.3 --- Intrinsic Viscosity --- p.8 / Chapter 1.3.4 --- Size Exclusion Chromatography (SEC) --- p.9 / Chapter 1.3.5 --- Mass Spectrometry --- p.10 / Chapter 1.4 --- Matrix-assisted Laser Desorption / Ionization Mass Spectrometry --- p.12 / Chapter 1.5 --- MALDI-TOF Mass Spectrometry of polysaccharides --- p.13 / Chapter 1.6 --- Outline of project --- p.15 / Chapter Chapter two --- Instrumental and experimental / Chapter 2.1 --- Instrumentation --- p.18 / Chapter 2.1.1 --- Laser-based ion source --- p.18 / Chapter 2.1.2 --- Time-of-flight (TOF) analyzer --- p.19 / Chapter 2.1.3 --- Ion deflector --- p.23 / Chapter 2.1.4 --- Detector and data acquisition system --- p.23 / Chapter 2.2 --- Experimental --- p.26 / Chapter 2.2.1 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by water extraction --- p.26 / Chapter 2.2.2 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by dimethyl ssulfoxide (DMSO) extraction --- p.26 / Chapter 2.2.3 --- Fractionation of water-soluble polysaccharides by Gel Permeation Chromatography (GPC) --- p.27 / Chapter 2.2.4 --- Bradford protein assay --- p.28 / Chapter 2.2.5 --- Phenol / sulfuric acid assay --- p.28 / Chapter 2.2.6 --- Sample preparation in MS --- p.28 / Chapter 2.2.7 --- Calibration of MALDI-TOF-MS --- p.29 / Chapter 2.2.8 --- Data analysis --- p.29 / Chapter Chapter three --- Extraction and purification of water-soluble polysaccharides from Ganoderma Lucidum / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Experimental --- p.32 / Chapter 3.2.1 --- Extraction efficiency --- p.32 / Chapter 3.2.2 --- Dialysis --- p.32 / Chapter 3.2.3 --- Signal suppression effect --- p.32 / Chapter 3.2.4 --- Sevag method --- p.33 / Chapter 3.2.5 --- Trichloroacetic acid (TCA) precipitation --- p.33 / Chapter 3.2.6 --- Bradford Protein Assay --- p.34 / Chapter 3.2.7 --- Phenol sulfuric acid assay --- p.34 / Chapter 3.3 --- Results and discussion --- p.35 / Chapter 3.3.1 --- Extraction efficiency --- p.35 / Chapter 3.3.2 --- Purification of crude polysaccharides --- p.37 / Chapter 3.3.3 --- Desalting --- p.38 / Chapter 3.3.4 --- Deproteination --- p.40 / Chapter 3.3.4.1 --- Monitoring of protein contents --- p.40 / Chapter 3.3.4.2 --- Monitoring of carbohydrate contents --- p.45 / Chapter 3.3.4.3 --- Deproteination using Sevag and TCA procipitation method --- p.47 / Chapter 3.4 --- Conclusions --- p.54 / Chapter Chapter four --- Evaluation of MW and MWD of water-soluble Polysaccharides extracted from Ganoderma Lucidum / Chapter 4.1 --- Introduction --- p.55 / Chapter 4.2 --- Experimental --- p.58 / Chapter 4.2.1 --- Aqueous DHB matrix --- p.58 / Chapter 4.2.2 --- Aqueous DHB/NH4F matrix --- p.58 / Chapter 4.2.3 --- Aqueous DHB matrix in TA solution --- p.58 / Chapter 4.2.4 --- Aqueous DHB/NH4F matrix in TA solution --- p.59 / Chapter 4.2.5 --- Aqueous DHB/NH4F matrix with sodium salt in TA solution --- p.59 / Chapter 4.2.6 --- Aqueous DHB/NH4F matrix with potassium salt in TA solution --- p.59 / Chapter 4.2.7 --- Fractionation of water-soluble polysaccharide extracted by DMSO by Gel Permeation Chromatography (GPC) --- p.59 / Chapter 4.2.8 --- Ultra-violet absorption spectrometry (UV-VIS) --- p.60 / Chapter 4.3 --- Results and discussion --- p.60 / Chapter 4.3.1 --- Development of matrix and solvent system for water-soluble polysaccharides --- p.60 / Chapter 4.3.2 --- MW and MWD of water-soluble polysaccharides extracted from Ganoderma Lucidum --- p.64 / Chapter 4.3.2.1 --- Water extraction --- p.65 / Chapter 4.3.2.2 --- DMSO extraction followed by water-back extraction --- p.70 / Chapter 4.4 --- Conclusions --- p.79 / Chapter Chapter five --- Concluding remarks --- p.82 / References --- p.84

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.

Thesis (M.Med.Sc.)--University of Hong Kong, 2003. / Includes bibliographical references (leaves 83-93). Also available in print.

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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

published_or_final_version / Paediatrics and Adolescent Medicine / Master / Master of Medical Sciences