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