Koo, Siu Chung Nicolas. / Thesis submitted in: November 2007. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 78-86). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Chinese Abstract --- p.v / Acknowledgements --- p.vi / Abbreviations --- p.viii / Table of contents --- p.xi / List of figures --- p.xv / List of tables --- p.xvii / Chapter 1. --- General Introduction / Chapter 1.1 --- General introduction to salt tolerance in plant --- p.1 / Chapter 1.1.1 --- Adverse effecst of high salinity in plant cells / Chapter 1.1.1.1 --- Ion toxicity --- p.1 / Chapter 1.1.1.2 --- Disturbed osmotic homeostasis --- p.2 / Chapter 1.1.1.3 --- Oxidative stress --- p.3 / Chapter 1.1.2 --- Major salt tolerance strategy in plant / Chapter 1.1.2.1 --- Maintenance of ion homeostasis --- p.4 / Chapter 1.1.2.2 --- Maintaining osmotic homeostasis --- p.4 / Chapter 1.1.2.3 --- Detoxification of Reactive oxygen species --- p.4 / Chapter 1.2 --- Cytosolic Calcium signal in plant / Chapter 1.2.1 --- General introduction of calcium in plant --- p.6 / Chapter 1.2.2 --- Calcium transport in plant cell --- p.7 / Chapter 1.2.3 --- Cytosolic calcium signals in plant under abiotic stress --- p.9 / Chapter 1.2.4 --- Responding to cytosolic calcium signals --- p.12 / Chapter 1.3 --- Calcium mediated ion homeostasis in plant under salt stress / Chapter 1.3.1 --- General introduction on Calcium dependent ion channels in plant --- p.13 / Chapter 1.3.2 --- SOS family cascade in Arabidopsis --- p.13 / Chapter 1.4 --- The interaction between cytosolic calcium and reactive oxygen species in plants --- p.14 / Chapter 1.5 --- "Calcium signaling mediated by Inositol 1,4,5 triphosphate in plant" --- p.15 / Chapter 1.6 --- Study on HAL2 and its homolog in plant --- p.18 / Chapter 1.7 --- Previous studies on GmSAL1 in Prof. Lam's lab --- p.20 / Chapter 1.8 --- Hypothesis and significant of this project --- p.21 / Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials / Chapter 2.1.1 --- "Plants, bacterial strains and vectors" --- p.23 / Chapter 2.1.2 --- Chemicals and Regents --- p.25 / Chapter 2.1.3 --- Commercial kits --- p.26 / Chapter 2.1.4 --- Primers and Adaptors --- p.27 / Chapter 2.1.5 --- Equipments and facilities used --- p.27 / Chapter 2.1.6 --- "Buffer, solution, gel and medium" --- p.27 / Chapter 2.1.7 --- Software --- p.28 / Chapter 2.2 --- Methods / Chapter 2.2.1 --- Molecular Techniques / Chapter 2.2.1.1 --- Bacterial cultures for recombinant DNA and plant transformation --- p.29 / Chapter 2.2.1.2 --- Recombinant DNA techniques --- p.29 / Chapter 2.2.1.3 --- Preparation and transformation of Agrobacterium competent cells --- p.30 / Chapter 2.2.1.4 --- Gel electrophoresis --- p.31 / Chapter 2.2.1.5 --- DNA and RNA extractions --- p.32 / Chapter 2.2.1.6 --- Generation of single-stranded DIG-labeled PCR probes --- p.34 / Chapter 2.2.1.7 --- Testing the concentration of DIG-labeled probes --- p.36 / Chapter 2.2.1.8 --- Northern blot analysis --- p.36 / Chapter 2.2.1.9 --- PCR techniques --- p.37 / Chapter 2.2.1.10 --- Sequencing --- p.38 / Chapter 2.2.2 --- Plant cell culture and transformation / Chapter 2.2.2.1 --- Arabidopsis thaliana --- p.39 / Chapter 2.2.2.2 --- Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cells --- p.39 / Chapter 2.2.3 --- Growth and treatment conditions for plants / Chapter 2.2.3.1 --- Growth and salt treatment condition of soybean samples for gene expression studies of GmSAL1 --- p.40 / Chapter 2.2.3.2 --- Root assay of GmSAL1l transgenic Arabidopsis thaliana --- p.41 / Chapter 2.2.4 --- "Cell viability, ROS detection and confocal microscopy" / Chapter 2.2.4.1 --- Cell viability assay --- p.42 / Chapter 2.2.4.2 --- Detection of intracellular contents of Na+ --- p.42 / Chapter 2.2.4.3 --- Detection of Reactive oxygen species (ROS) --- p.42 / Chapter 2.2.4.4 --- Confocal microscopy --- p.43 / Chapter 2.2.4.5 --- Images processing and analysis --- p.43 / Chapter 2.2.5 --- Statistical analysis --- p.44 / Chapter 3 --- Results / Chapter 3.1 --- GmSAL1 sequence analysis --- p.45 / Chapter 3.2 --- Expression of GmSAL1 was induced by NaCl stress --- p.49 / Chapter 3.3 --- Construction of GmSAL1 transgenic tobacco BY-2 cell line --- p.50 / Chapter 3.4 --- Ectopic expression of GmSAL1 alleviates NaCl stress in transgenic tobacco BY-2 cells --- p.52 / Chapter 3.5 --- GmSAL1 enhances vacuolar sodium compartmentalization in transgenic tobacco BY-2 cell under NaCl treatment --- p.55 / Chapter 3.6 --- GmSAL1 helps maintain cell turgidity in transgenic tobacco BY-2 cell under NaCl treatment --- p.58 / Chapter 3.7 --- GmSAL1 enhances ROS scavenging in transgenic tobacco BY-2 cell under NaCl treatment --- p.61 / Chapter 3.8 --- Effect of expressing GmSAL1 in Arabidopsis thaliana under salt stress --- p.64 / Chapter 4 --- Discussion --- p.66 / Chapter 4.1 --- Sequence analysis and enzyme activity of GmSAL1 --- p.68 / Chapter 4.2 --- Gene expression profile of GmSAL1 --- p.70 / Chapter 4.3 --- Functional analysis of GmSAL1 in transgenic tobacco BY-2 cells / Chapter 4.3.1 --- GmSAL1 protects transgenic BY-2 cells under salt treatment --- p.71 / Chapter 4.3.2 --- GmSAL1 regulates Na+ compartmentalization and ROS scavenging in transgenic BY-2 cells under NaCl treatment in a calcium dependent manner --- p.72 / Chapter 4.4 --- Functional tests of GmSAL1 transgenic A. thaliana --- p.75 / Chapter 5 --- Conclusion and perspective --- p.76 / References --- p.78 / "Appendix I: Substrate specificity and Km, Kcat values of GmSAL1 protein" --- p.87 / Appendix II: Restriction and modifying enzymes --- p.89 / Appendix II: Chemicals --- p.90 / Appendix III: Commercial kits --- p.94 / Appendix IV: Equipments and facilities used --- p.95 / "Appendix V: Buffer, solution, gel and medium formulation" --- p.96
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326174 |
| Date | January 2008 |
| Contributors | Koo, Siu Chung Nicolas., Chinese University of Hong Kong Graduate School. Division of Biology. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvi, 98 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use 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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