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GmSAL1 enhances vacuolar sodium ion compartmentalization and ROS scavenging in a calcium dependent manner.

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

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326174
Date January 2008
ContributorsKoo, Siu Chung Nicolas., Chinese University of Hong Kong Graduate School. Division of Biology.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xvi, 98 leaves : ill. (some col.) ; 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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