by Lau Tai Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 117-127). / Abstracts in English and Chinese. / Table of Content --- p.i / Abbreviations --- p.iv / Abstract --- p.v / List of Figures --- p.ix / List of Tables --- p.xii / Chapter 1. --- Chapter One: Introduction --- p.1 / Chapter 1.1 --- Background on orchids --- p.2 / Chapter 1.2 --- Background on Dendrobium --- p.7 / Chapter 1.3 --- Background and history on Herba Dendrobii --- p.9 / Chapter 1.4 --- Reasons for study of Herba Dendrobii --- p.12 / Chapter 1.4.1 --- Demand --- p.12 / Chapter 1.4.2 --- Adulteration --- p.13 / Chapter 1.4.3 --- CITES --- p.13 / Chapter 1.5 --- Scientific researches on Herba Dendrobii --- p.14 / Chapter 1.5.1 --- Morphological studies --- p.15 / Chapter 1.5.2 --- Anatomical and microscopic studies --- p.16 / Chapter 1.5.3 --- Phytochemistry --- p.20 / Chapter 1.5.3.1 --- Chemicals identified --- p.20 / Chapter 1.5.3.2 --- Chemical authentication of Herba Dendrobii --- p.23 / Chapter 1.5.3.3 --- Effect of treatment on chemical composition --- p.23 / Chapter 1.5.4 --- Phylogenetic study of Dendrobium --- p.25 / Chapter 1.5.4.1 --- Phylogenetic analysis by molecular methods --- p.25 / Chapter 1.5.4.2 --- Phylogenetic analysis by anatomical methods --- p.27 / Chapter 1.5.5 --- Pharmacological effect --- p.29 / Chapter 2. --- Chapter two: Objectives and strategies --- p.30 / Chapter 3. --- Chapter Three: Materials and Methods --- p.33 / Chapter 3.1 --- Source of samples and their treatment --- p.34 / Chapter 3.1.1 --- Fresh materials --- p.34 / Chapter 3.1.2 --- Dry materials --- p.34 / Chapter 3.1.3 --- Outgroup species --- p.35 / Chapter 3.2 --- Experimental protocol --- p.40 / Chapter 3.2.1 --- Rationale of the experiment --- p.40 / Chapter 3.2.2 --- DNA extraction --- p.41 / Chapter 3.2.2.1 --- Cetyltrimethylammonium bromide extraction method --- p.41 / Chapter 3.2.2.1a --- Reagents and buffers --- p.41 / Chapter 3.2.2.1b --- Procedures of CTAB extraction method --- p.42 / Chapter 3.2.2.2 --- Modified DNA isolation protocol for dry samples --- p.43 / Chapter 3.2.2.2a --- Reagents and buffers --- p.43 / Chapter 3.2.2.2b --- Procedures of modified DNA isolation protocol for dry plant samples --- p.44 / Chapter 3.2.3 --- Agarose gel electrophoresis of genomic DNA or PCR products --- p.45 / Chapter 3.2.3a --- Reagents and buffers --- p.45 / Chapter 3.2.3b --- Procedures of agarose gel electrophoresis of genomic DNA or PCR products --- p.45 / Chapter 3.2.4 --- Qualitative and quantitative analysis of DNA --- p.46 / Chapter 3.2.5 --- Amplification of the internal transcribed spacer 2 (ITS 2) region by Polymerase Chain Reaction --- p.47 / Chapter 3.2.5a --- Internal transcribed spacer 2 (ITS 2) region --- p.47 / Chapter 3.2.5b --- Procedures of polymerase chain reaction of ITS 2 region --- p.48 / Chapter 3.2.6 --- Purification of PCR products or cycle sequencing products --- p.48 / Chapter 3.2.6.1 --- Ethanol precipitation --- p.48 / Chapter 3.2.6.2 --- GENECLEAN® protocols --- p.49 / Chapter 3.2.6.3 --- Spin Column Purification --- p.49 / Chapter 3.2.7 --- Cycle Sequencing --- p.50 / Chapter 3.2.8 --- Sample Electrophoresis --- p.51 / Chapter 3.2.8a --- Equipment and reagents --- p.51 / Chapter 3.2.8b --- Procedures of sample electrophoresis --- p.52 / Chapter 3.2.9 --- Sequence analysis --- p.52 / Chapter 4. --- Results --- p.53 / Chapter 4.1 --- Fresh materials --- p.54 / Chapter 4.1.1 --- Genomic DNA --- p.54 / Chapter 4.1.2 --- PCR products --- p.59 / Chapter 4.1.3 --- Sequence alignment --- p.66 / Chapter 4.1.4 --- Comparison of the sequences --- p.94 / Chapter 4.1.5 --- Percentage difference among Dendrobium --- p.96 / Chapter 4.1.6 --- Intra-specific variation of orchid species --- p.96 / Chapter 4.1.7 --- Phylogenetic analysis --- p.99 / Chapter 4.2 --- Dry materials --- p.101 / Chapter 4.2.1 --- Genomic DNA --- p.101 / Chapter 4.2.2 --- PCR products --- p.101 / Chapter 4.2.3 --- Sequencing result --- p.101 / Chapter 5. --- Discussion and Conclusion --- p.107 / Chapter 5.1 --- Reasons for authentication of Herba Dendrobii --- p.108 / Chapter 5.2 --- Fresh materials of Herba Dendrobii --- p.109 / Chapter 5.2.1 --- Authentication --- p.109 / Chapter 5.2.2 --- Phylogenetic analysis --- p.111 / Chapter 5.3 --- Dry materials of Herba Dendrobii --- p.114 / Chapter 5.4 --- Evaluation of the experimental method --- p.115 / Chapter 5.5 --- Conclusion --- p.116 / Chapter 6. --- Reference --- p.117 / Chapter 7. --- Appendix / Appendix 1: Number of species in each medicinal orchid geneus --- p.128 / Appendix 2: Photographs showing 15 of the 17 species of orchids used in this research project --- p.131

University of Macau / Institute of Chinese Medical Sciences