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Establishment of cell culture and characterization of seed coat pigments of vigna sinensis.

Yip Mei-kuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 93-102). / Abstracts in English and Chinese. / Acknowledgments --- p.i / List of abbreviations --- p.ii / Abstract --- p.iii / Table of Contents --- p.vi / List of tables --- p.x / List of figures --- p.xii / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Plant of interest --- p.1 / Chapter 1.2 --- Literature review --- p.2 / Chapter 1.2.1 --- Anthocyanins-natural pigments in plants --- p.2 / Chapter 1.2.1.1 --- Sources and biosynthesis --- p.2 / Chapter 1.2.1.2 --- Chemical properties --- p.2 / Chapter 1.2.1.3 --- Biological effects --- p.3 / Chapter 1.2.2 --- Characterization of anthocyanins --- p.4 / Chapter 1.2.3 --- Plant tissue and cell cultures --- p.6 / Chapter 1.2.4 --- Induction of anthocyanins in plant tissue culture --- p.7 / Chapter 1.2.5 --- Factors affecting anthocyanin production --- p.8 / Chapter 1.2.5.1 --- Plant hormones --- p.8 / Chapter 1.2.5.2 --- Nutrients --- p.9 / Chapter 1.2.5.2.1 --- Phosphate --- p.9 / Chapter 1.2.5.2.2 --- Nitrogen --- p.9 / Chapter 1.2.5.3 --- Osmoticums --- p.10 / Chapter 1.2.5.3.1 --- Sucrose --- p.10 / Chapter 1.2.5.3.2 --- Other factors --- p.10 / Chapter 1.3 --- Research objectives --- p.12 / Chapter 2. --- Materials and methods --- p.16 / Chapter 2.1 --- Plant materials --- p.16 / Chapter 2.2 --- Study of pigment formation at different developmental stages --- p.16 / Chapter 2.2.1 --- Cultivation of Vigna sinensis --- p.16 / Chapter 2.2.2 --- Sample collection --- p.16 / Chapter 2.2.3 --- HPLC analysis of pigmented vegetative tissues --- p.16 / Chapter 2.2.4 --- HPLC analysis of seed coats at different developmental stages --- p.17 / Chapter 2.3 --- Characterization of seed coat pigments --- p.17 / Chapter 2.3.1 --- Extraction of seed coats pigments --- p.17 / Chapter 2.3.2 --- Acid hydrolysis of anthocyanins --- p.17 / Chapter 2.3.3 --- High performance liquid chromatography --- p.18 / Chapter 2.3.3.1 --- HPLC system --- p.18 / Chapter 2.3.3.2 --- Analytical conditions --- p.18 / Chapter 2.4 --- Establishment of tissue culture system --- p.19 / Chapter 2.4.1 --- Aseptic plant stocks --- p.19 / Chapter 2.4.2 --- Shoot-tip cultures --- p.19 / Chapter 2.4.3 --- Callus initiation --- p.19 / Chapter 2.4.3.1 --- From seed coats --- p.20 / Chapter 2.4.3.2 --- From vegetative tissues --- p.20 / Chapter 2.4.3.3 --- Light and dark --- p.20 / Chapter 2.4.4 --- Optimization of callus growth --- p.21 / Chapter 2.4.4.1 --- Basal medium --- p.21 / Chapter 2.4.4.2 --- Combination of various plant hormones --- p.21 / Chapter 2.4.4.3 --- Basal salt --- p.21 / Chapter 2.5 --- Studies of anthocyanin production in hypocotyl callus cultures --- p.22 / Chapter 2.5.1 --- Effects of nutrients --- p.22 / Chapter 2.5.1.1 --- Nitrogen --- p.22 / Chapter 2.5.1.2 --- Phosphate --- p.22 / Chapter 2.5.2 --- Osmotic stress --- p.22 / Chapter 2.5.2.1 --- Sucrose --- p.22 / Chapter 2.5.2.2 --- Mannitol --- p.23 / Chapter 2.5.2.3 --- Sodium chloride --- p.23 / Chapter 2.5.2.4 --- Polyethylene glycol --- p.23 / Chapter 2.6 --- Studies of anthocyanin production in cell suspension cultures --- p.23 / Chapter 2.6.1 --- Effects of nutrients --- p.24 / Chapter 2.6.1.1 --- Nitrogen --- p.24 / Chapter 2.6.1.2 --- Phosphate --- p.24 / Chapter 2.6.2 --- Osmotic stress --- p.25 / Chapter 2.6.2.1 --- Sucrose --- p.25 / Chapter 2.6.2.2 --- Polyethylene glycol --- p.25 / Chapter 2.6.3 --- Effects of other factors --- p.25 / Chapter 2.6.3.1 --- Riboflavin --- p.25 / Chapter 2.6.3.2 --- pH --- p.26 / Chapter 2.7 --- Measurement of cell growth --- p.26 / Chapter 2.8 --- Estimation of anthocyanins --- p.26 / Chapter 2.9 --- Statistical analysis --- p.27 / Chapter 3. --- Results --- p.30 / Chapter 3.1 --- Study of pigment formation at different developmental stages --- p.30 / Chapter 3.1.1 --- General description --- p.30 / Chapter 3.1.2 --- HPLC analysis of developing seed coats and other vegetative tissues --- p.30 / Chapter 3.1.3 --- The relationship between pigment formation and seed development --- p.30 / Chapter 3.2 --- Characterization of seed coat pigments --- p.31 / Chapter 3.3 --- Establishment of tissue culture system --- p.43 / Chapter 3.3.1 --- Callus initiations from seed coats --- p.43 / Chapter 3.3.2 --- Callus initiations from vegetative tissues --- p.43 / Chapter 3.3.3 --- Optimization of callus growth --- p.43 / Chapter 3.3.3.1 --- Effects of NAA and BA --- p.43 / Chapter 3.3.3.2 --- Effects of basal medium and combinations of plant hormones --- p.44 / Chapter 3.3.3.3 --- Effects of basal salt --- p.44 / Chapter 3.4 --- Studies of anthocyanin production in hypocotyl callus culture --- p.54 / Chapter 3.4.1 --- Effects of nutrients --- p.54 / Chapter 3.4.1.1 --- Effects of total nitrogen --- p.54 / Chapter 3.4.1.2 --- Effects of phosphate --- p.54 / Chapter 3.4.2 --- Effects of plant hormones --- p.55 / Chapter 3.4.3 --- Osmotic stress --- p.55 / Chapter 3.5 --- Establishment of suspension culture system --- p.64 / Chapter 3.6 --- Studies of anthocyanin production in seed coat suspension cultures --- p.64 / Chapter 3.6.1 --- Nutrient effects on suspension cultures --- p.64 / Chapter 3.6.2 --- Osmotic stress on suspension cultures --- p.65 / Chapter 3.6.3 --- Effects of phosphate with high nitrogen --- p.65 / Chapter 3.6.4 --- Effects of riboflavin with high nitrogen --- p.66 / Chapter 3.6.5 --- Influence of pH with high nitrogen --- p.66 / Chapter 4. --- Discussion --- p.79 / Chapter 4.1 --- Anthocyanin in vegetative tissues and seed coats of Vigna sinensis --- p.79 / Chapter 4.2 --- Factors affecting callus initiation in Vigna sinensis --- p.81 / Chapter 4.2.1 --- Explant types --- p.81 / Chapter 4.2.2 --- Plant hormones --- p.82 / Chapter 4.2.3 --- Basal medium --- p.82 / Chapter 4.3 --- Factors affecting anthocyanin production in callus cultures derived from hypocotyls --- p.83 / Chapter 4.3.1 --- Nutrients --- p.83 / Chapter 4.3.2 --- Osmotic stress --- p.85 / Chapter 4.4 --- Factors affecting anthocyanin production in suspension culture derived from seed coats --- p.86 / Chapter 4.4.1 --- Nutrients --- p.86 / Chapter 4.4.2 --- Osmotic stress --- p.87 / Chapter 4.5 --- Comparison of anthocyanin production from natural source and plant tissue cultures of V.sinensis --- p.89 / Chapter 4.6 --- Further studies --- p.89 / Chapter 5. --- Conclusions --- p.91 / References --- p.93

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323181
Date January 2000
ContributorsYip, Mei-kuen., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xiii, 102 leaves : ill. ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

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