Molecular authentication and taxonomy of radix stemonae.

Chan Yiu-Man.Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.Includes bibliographical references (leaves 123-127).Abstracts in English and Chinese.Abstract (in English) --- p.iAbstract (in Chinese) --- p.iiiAcknowledgements --- p.ivContents --- p.vList of Figures --- p.ixList of Tables --- p.xiAbbreviations --- p.xiiChapter Chapter 1. --- Introduction --- p.1Chapter 1.1 --- Background --- p.1Chapter 1.1.1 --- Source plants of Radix Stemonae --- p.1Chapter 1.1.2 --- Medicinal usage of Radix Stemonae --- p.1Chapter 1.1.3 --- Stemonaceae --- p.2Chapter 1.1.4 --- Stemonaceae of China --- p.3Chapter 1.1.5 --- Circumscriptions of Stemonaceae --- p.3Chapter 1.1.6 --- Affinity of Stemonaceae --- p.7Chapter 1.2 --- Molecular Markers for Phylogenetic study --- p.10Chapter 1.2.1 --- Choosing appropriate DNA region(s) --- p.10Chapter 1.2.2 --- Chloroplast DNA markers --- p.10Chapter 1.2.3 --- Nuclear sequences --- p.11Chapter 1.3 --- Objectives --- p.13Chapter Chapter 2. --- Materials and Methods --- p.14Chapter 2.1 --- Sources of Samples and Their Treatment --- p.14Chapter 2.1.1 --- Fresh Materials --- p.14Chapter 2.1.2 --- DNA Samples --- p.15Chapter 2.1.3 --- Dried Medicinal Material from Commerical Market --- p.15Chapter 2.2 --- DNA Isolation from Plant Materials --- p.20Chapter 2.2.1 --- Reagents for DNA Isolation --- p.20Chapter 2.2.2 --- Procedures of DNA Isolation --- p.22Chapter 2.2.2.1 --- Treatments of Plant Materials --- p.22Chapter 2.2.2.2 --- CTAB (Cetyltrimethylammonium bromide) Method --- p.23Chapter 2.2.2.3 --- DNeasy ® Plant Mini Kit --- p.24Chapter 2.2.2.4 --- GenElute Plant Genomic DNA Miniprep --- p.24Chapter 2.2.2.5 --- Extraction method of Kang et. al (1998) --- p.25Chapter 2.2.2.6 --- Agarose Gel Electrophoresis of Genomic DNA --- p.26Chapter 2.3 --- Polymerase Chain Reaction (PCR) --- p.27Chapter 2.3.1 --- Reagents --- p.27Chapter 2.3.2 --- Procedures --- p.28Chapter 2.4 --- "Ligation, Transformation and Bacterial Culture for 5S rRNA Spacer Analysis" --- p.30Chapter 2.4.1 --- Reagents --- p.30Chapter 2.4.2 --- Procedures --- p.33Chapter 2.4.2.1 --- Ligation --- p.33Chapter 2.4.2.2 --- Transformation --- p.33Chapter 2.4.2.3 --- Blue-White Screening --- p.33Chapter 2.4.2.4 --- Plasmid Isolation --- p.34Chapter 2.4.2.5 --- Screening of plasmid DNA by PCR --- p.35Chapter 2.5 --- Cycle Sequencing and Electrophoresis --- p.36Chapter 2.5.1 --- Instruments and Reagents --- p.36Chapter 2.5.2 --- Procedures of Cycle Sequencing and Electrophoresis --- p.37Chapter 2.5.2.1 --- Cycle sequencing --- p.37Chapter 2.5.2.2 --- Ethanol Precipitation --- p.38Chapter 2.5.2.3 --- Electrophoresis --- p.38Chapter 2.6 --- Sequence Analysis --- p.40Chapter Chapter 3. --- Taxonomic Study of Chinese Stemona species --- p.41Chapter 3.1 --- History of the Genus Stemona --- p.41Chapter 3.2 --- Characteristics of the Genus Stemona --- p.42Chapter 3.3 --- Characteristics of Stemona sessilifolia (Miquel) Miquel (including Stemona shandongensis D. K. Zang) --- p.45Chapter 3.4 --- Characteristics of Stemona japonica (Blume) Miquel --- p.52Chapter 3.5 --- Characteristics of Stemona tuberosa Loureiro --- p.55Chapter 3.6 --- Characteristics of Stemona parviflora C. H. Wright --- p.61Chapter 3.7 --- Characteristics of Stemona mairei (H. Leveille) K. Krause --- p.65Chapter 3.8 --- Characteristics of Stemona kerrii Craib --- p.67Chapter Chapter 4. --- DNA Sequence Analysis for Authentication and Systematics --- p.69Chapter 4.1 --- DNA Extraction --- p.70Chapter 4.2 --- PCR --- p.73Chapter 4.3 --- DNA Authentication of Radix Stemonae --- p.77Chapter 4.3.1 --- TrnL intron sequences --- p.77Chapter 4.3.2 --- 5S rRNA spacer sequences --- p.86Chapter 4.3.3 --- Conclusion of DNA Authentication --- p.107Chapter 4.4 --- Molecular Systematics Analysis --- p.108Chapter 4.4.1 --- Circumscription of Stemonaceae and its affinity to other monocots based on trnL intron sequences --- p.109Chapter 4.4.2 --- Interspecific relationship of Stemona --- p.114Chapter Chapter 5. --- Discussion --- p.116Chapter 5.1 --- Molecular Authentication of Radix Stemonae --- p.116Chapter 5.2 --- Molecular Markers --- p.117Chapter 5.3 --- The Variation in Stemona tuberosa --- p.117Chapter 5.4 --- Comparsion of Stemona sessilifolia and S. shandongensis --- p.118Chapter 5.5 --- Circumscription of Stemonaceae --- p.119Chapter 5.6 --- Affinity of Stemonaceae --- p.120Chapter Chapter 6. --- Conclusion --- p.122References --- p.12