Molecular authentication of the traditional Chinese medicine Fructus Evodiae and systematics of Rutaceae.

Poon Wing-sem.Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.Includes bibliographical references (leaves 163-171).Abstracts in English and Chinese.ABSTRACT I-IV --- p.I-IVACKNOWLEDGMENTS V --- p.VTABLE OF CONTENT --- p.VI-VIIILIST OF FIGURES AND TABLES --- p.IX-XILIST OF ABBREVIATIONS --- p.XIIChapter CHAPTER ONE --- LITERATURE REVIEW --- p.1Chapter 1.1 --- Rutaceae --- p.1Chapter 1.1.1 --- Introduction --- p.1Chapter 1.1.2 --- taxonomy of Rutaceae --- p.2Chapter 1.1.3 --- Controversial taxonomic issues --- p.4Chapter 1.1.3.1 --- Subfamilies Rutoideae and Toddalioideae --- p.4Chapter 1.1.3.2 --- "Euodia, Melicope and Tetradium" --- p.7Chapter 1.1.3.2.1 --- History --- p.7Chapter 1.1.3.2.2 --- Arguments based on morphology --- p.10Chapter 1.2 --- Molecular Approach --- p.12Chapter 1.2.1 --- Introduction to molecular systematics --- p.12Chapter 1.2.2 --- DNA sequence markers --- p.14Chapter 1.2.3 --- Applications --- p.18Chapter 1.3 --- Traditional Chinese Medicine (TCM) --- p.21Chapter 1.3.1 --- Introduction --- p.21Chapter 1.3.2 --- Fructus Evodiae --- p.22Chapter 1.3.3 --- Functional chemicals and pharmacological effects of Fructus Evodiae --- p.23Chapter 1.3.4 --- Problem in authentication --- p.25Chapter 1.4 --- Objectives --- p.27Chapter CHAPTER TWO --- METHODOLOGY AND MATERIALS --- p.29Chapter 2.1 --- Plant and Herb Materials --- p.29Chapter 2.2 --- DNA extraction --- p.44Chapter 2.2.1 --- Modified cetyltriethylammonium bromide (CTAB) extraction --- p.44Chapter 2.2.2 --- Kit extraction --- p.45Chapter 2.2.2.1 --- DNeasy® Plant MiniKit of Qiagen --- p.45Chapter 2.2.2.2 --- GenElute´ёØ Plant Genomic DNA Miniprep Kit of Sigma® --- p.46Chapter 2.3 --- Polymerase chain reaction (PCR) reaction --- p.47Chapter 2.4 --- DNA gel electrophoresis --- p.49Chapter 2.5 --- PCR product purification --- p.49Chapter 2.5.1 --- Rapid Gel Extraction System of Marligen Biosciences INC --- p.50Chapter 2.5.2 --- Gel-M´ёØ Gel Extraction System --- p.50Chapter 2.6 --- Ligation and transformation --- p.51Chapter 2.6.1 --- Ligation and transformation --- p.51Chapter 2.6.2 --- Cell culture --- p.52Chapter 2.6.3 --- Plasmid extraction --- p.52Chapter 2.7 --- Determination of DNA concentration --- p.54Chapter 2.8 --- Cycle Sequencing --- p.54Chapter 2.9 --- Sequence Analysis --- p.55Chapter 2.10 --- Materials --- p.56Chapter CHAPTER THREE --- MOLECULAR AUTHENTICATION OF FRUCTUSEVODIAE --- p.60Chapter 3.1 --- Results and data analysis --- p.60Chapter 3.1.1 --- Authentication based on ITS-1 region --- p.60Chapter 3.1.1.1 --- Phylogram study --- p.60Chapter 3.1.1.2 --- Sequence alignment --- p.65Chapter 3.1.1.3 --- ITS-1 region nucleotide differences significant in authentication of Fructus Evodiae --- p.71Chapter 3.1.1.4 --- Comparison of sequences --- p.74Chapter 3.1.2 --- Authentication based on ITS-2 region --- p.78Chapter 3.1.2.1 --- Phylogram study --- p.78Chapter 3.1.2.2 --- Sequence alignment --- p.82Chapter 3.1.2.3 --- ITS-2 region nucleotide differences significant inauthentication of Fructus Evodiae --- p.86Chapter 3.1.2.4 --- Comparison of sequences --- p.89Chapter 3.2 --- Discussion --- p.93Chapter 3.2.1 --- Molecular markers --- p.93Chapter CHAPTER FOUR --- PHYLOGENETIC STUDIES ON RUTACEAE --- p.96Chapter 4.1 --- Results and data analysis --- p.96Chapter 4.1.1 --- Chloroplast trnL intron region --- p.96Chapter 4.1.1.1 --- Sequence alignment --- p.96Chapter 4.1.1.2 --- Phylogenetic analysis --- p.107Chapter 4.1.2 --- Chloroplast trnL-F intergenic spacer region --- p.116Chapter 4.1.2.1 --- Sequence alignment --- p.116Chapter 4.1.2.2 --- Phylogenetic analysis --- p.126Chapter 4.1.3 --- Nuclear ITS-1 region --- p.132Chapter 4.1.3.1 --- Sequence alignment --- p.132Chapter 4.1.3.2 --- Phylogenetic analysis --- p.143Chapter 4.2 --- Discussion --- p.152Chapter 4.2.1 --- "Euodia, Melicope and Tetradium" --- p.152Chapter 4.2.2 --- Tetradium --- p.153Chapter 4.2.3 --- Tetradium and Phellodendron --- p.155Chapter 4.2.4 --- Zanthoxylum and Toddalia --- p.156Chapter 4.2.5 --- Rutoideae and Toddalioideae --- p.156Chapter 4.2.6 --- Tree constructing methods --- p.158Chapter CHAPTER FIVE --- CONCLUSION --- p.161REFERENCES --- p.16