Wong, Cheuk Hon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 69-75). / Abstracts in English and Chinese. / Thesis Committee --- p.ii / Statement --- p.iii / Abstract --- p.iv / 摘要 --- p.V / Acknowledgements --- p.vi / General Abbreviations --- p.viii / Abbreviations of Chemicals --- p.X / List of Figures --- p.xi / List of Tables --- p.xii / Table of Contents --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Food insecurity and abiotic stress --- p.1 / Chapter 1.2 --- Overview of the molecular basis of salt tolerance in plants --- p.4 / Chapter 1.2.1 --- Stress perception --- p.4 / Chapter 1.2.2 --- Signal transduction --- p.5 / Chapter 1.2.2.1 --- ABA dependent pathway --- p.6 / Chapter 1.2.2.2 --- ABA independent pathway --- p.8 / Chapter 1.2.2.3 --- The SOS pathway --- p.9 / Chapter 1.2.3 --- Abiotic stress tolerance mechanisms --- p.9 / Chapter 1.2.3.1 --- Ion transporters --- p.10 / Chapter 1.2.3.2 --- Osmolytes --- p.10 / Chapter 1.2.3.3 --- Antioxidants and ROS scavenging genes --- p.11 / Chapter 1.2.3.4 --- Late Embryogenesis Abundant proteins --- p.12 / Chapter 1.2.3.5 --- "Heat shock factor, heat shock element and heat shock protein" --- p.12 / Chapter 1.2.3.5.1 --- Hsp families --- p.14 / Chapter 1.2.3.5.2 --- Hsp network --- p.18 / Chapter 1.3 --- Pervious studies on GmDNJl --- p.19 / Chapter 1.4 --- "Hypothesis, significances and objectives of this project" --- p.19 / Chapter Chapter 2 --- Materials and Methods --- p.21 / Chapter 2.1 --- Materials --- p.21 / Chapter 2.1.1 --- "Plants, strains, and vectors" --- p.21 / Chapter 2.1.2 --- Chemicals and reagents --- p.24 / Chapter 2.1.3 --- Commercial kits --- p.25 / Chapter 2.1.4 --- Primers --- p.26 / Chapter 2.1.5 --- Equipment and facilities --- p.27 / Chapter 2.1.6 --- Sequencing --- p.27 / Chapter 2.1.7 --- "Buffer, solution and medium" --- p.27 / Chapter 2.1.8 --- Software --- p.28 / Chapter 2.2 --- Methods --- p.29 / Chapter 2.2.1 --- Growth conditions of bacterial and yeast cultures --- p.29 / Chapter 2.2.2 --- Growth conditions and multiple abiotic stress treatments of Glycine max for GmDNJl expression study --- p.29 / Chapter 2.2.2.1 --- "RNA extraction, cDNA synthesis of real-time PCR" --- p.30 / Chapter 2.2.3 --- Growth conditions and multiple abiotic stress treatments of Arabidopsis thaliana for root length essay --- p.30 / Chapter 2.2.3.1 --- Surface sterilization of Arabidopsis thaliana seeds --- p.30 / Chapter 2.2.3.2 --- Abiotic stress treatments of Arabidopsis thaliana for root length essay --- p.31 / Chapter 2.2.4 --- Multiple abiotic stress treatments of Tobacco Bright Yellow-2 (BY2) cell for cell viability assay --- p.31 / Chapter 2.2.4.1 --- Construction of transgenic GmDNJl BY2 cell lines --- p.31 / Chapter 2.2.4.2 --- DNA extraction of transgenic GmDNJl BY2 cell lines by CTAB method --- p.32 / Chapter 2.2.4.3 --- PCR screening of transgenic GmDNJl BY2 cell lines --- p.33 / Chapter 2.2.4.4 --- Gene expression checking of GmDNJl in transgenic BY2 cell lines by real-time PCR --- p.34 / Chapter 2.2.4.4.1 --- RNA extraction of BY2 cell --- p.35 / Chapter 2.2.4.4.2 --- cDNA synthesis of BY2 cell RNA --- p.35 / Chapter 2.2.4.4.3 --- Real-time PCR ofBY2 cell cDNA --- p.36 / Chapter 2.2.4.5 --- Multiple abiotic stress treatments of BY2 cell lines for cell viability assay --- p.37 / Chapter 2.2.5 --- Yeast two-hybrid experiment --- p.37 / Chapter 2.2.5.1 --- Subcloning oi GmDNJl into pGBKT7 --- p.37 / Chapter 2.2.5.2 --- Transformation of pGBKT7 into Y187 --- p.38 / Chapter 2.2.5.3 --- Toxicity test of GAL4 GmDNJl fusion protein in Y187 --- p.38 / Chapter 2.2.5.4 --- Western blot analysis to check the expression of GAL4-GmDNJl in Y187 --- p.39 / Chapter 2.2.5.5 --- Screening of GmDNJl interacting protein partners by yeast mating --- p.39 / Chapter 2.2.5.6 --- Identification of positive interaction partners by retrieving plasmid from diploid yeast --- p.40 / Chapter Chapter 3 --- Results --- p.41 / Chapter 3.1 --- "Expression of GmDNJl was induced under dehydration stress, NaHCOs stress, oxidative stress and heat stress" --- p.41 / Chapter 3.2 --- Ectopic expression of GmDNJl in transgenic Arabidopsis does not confer extra tolerance under multiple stresses in root length essay --- p.44 / Chapter 3.3 --- Construction of transgenic GmDNJl BY2 cell --- p.47 / Chapter 3.3.1 --- PCR screening of GmDNJl in transgenic BY2 cell lines --- p.47 / Chapter 3.3.2 --- Gene expression checking of GmDNJl in transgenic BY2 cell lines by real-time PCR --- p.49 / Chapter 3.3.3 --- Ectopic expression of GmDNJl in transgenic BY2 cell lines do not enhance tolerance under multiple stresses in cell viability essay --- p.51 / Chapter 3.4 --- Yeast two-hybrid experiment --- p.53 / Chapter 3.4.1 --- Subcloning of pGBKTl-GmDNJl --- p.54 / Chapter 3.4.2 --- Toxicity test of GAL4 DNA BD-c-Myc-GmDNJ 1 fusion protein inY187 --- p.54 / Chapter 3.4.3 --- Western blotting showing expression of GAL4 DNA BD -c-Myc-GmDNJl in Y187 --- p.55 / Chapter 3.4.4 --- Screening of GmDNJl putative protein interacting partners by yeast mating --- p.56 / Chapter Chapter 4 --- Discussion --- p.60 / Chapter 4.1 --- "Expression of GmDNJl was induced under dehydration stress, NaHCOs stress, oxidative stress and heat stress" --- p.60 / Chapter 4.2 --- Ectopic expression of GmDNJl in transgenic Arabidopsis and BY2 cells did not confer extra tolerance under multiple stresses in root length essay --- p.63 / Chapter 4.3 --- Yeast two-hybrid experiment --- p.64 / Chapter 4.4 --- Putative function of GmDNJl --- p.65 / Chapter 4.5 --- Future Perspectives --- p.66 / Chapter Chapter 5 --- Conclusion --- p.68 / Chapter Chapter 6 --- References --- p.69 / Chapter Appendix I --- Chemical and reagents --- p.76 / Chapter Appendix II --- "Buffer, solution, gel and medium formulation" --- p.78 / Chapter Appendix III --- Supplementary results --- p.81
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_327570 |
| Date | January 2011 |
| Contributors | Wong, Cheuk Hon., Chinese University of Hong Kong Graduate School. Division of Life Sciences. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xviii, 82 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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