Chow, Cheung-ming. / Thesis submitted in: December 2002. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 149-155). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgement --- p.vi / General abbreviations --- p.viii / Abbreviations of chemicals --- p.ix / List of figures --- p.x / List of tables --- p.xiv / Table of contents --- p.xv / Chapter 1. --- Literature review --- p.1-26 / Chapter 1.1 --- General introduction of nitrogen metabolism in plants --- p.1 / Chapter 1.2 --- Interaction between nitrogen metabolism and other metabolic and developmental pathways in plants --- p.2 / Chapter 1.2.1 --- Carbon metabolism --- p.2 / Chapter 1.2.2 --- Development --- p.2 / Chapter 1.2.3 --- Flowering --- p.3 / Chapter 1.3 --- Metabolic signalling in plants --- p.5 / Chapter 1.3.1 --- Nitrogen signalling in plants --- p.5 / Chapter 1.3.1.1 --- Inorganic N signalling --- p.5 / Chapter 1.3.1.2 --- Organic N signalling --- p.6 / Chapter 1.3.2 --- Carbon signalling --- p.7 / Chapter 1.3.2.1 --- Signalling pathways --- p.7 / Chapter 1.3.2.2 --- Gene expression regulated by sugar --- p.8 / Chapter 1.3.2.3 --- Role of sugar signalling in growth and development --- p.9 / Chapter 1.4 --- Ways to elucidate a new signal transduction pathway --- p.10 / Chapter 1.4.1 --- Carbon signalling as a paradigm to provide hints for exploring nitrogen signalling in plants --- p.10 / Chapter 1.4.2 --- Designing long-term approach to tackle nitrogen signalling in plants --- p.14 / Chapter 1.5 --- "Molecular tools available to change the ""signal"" and the proposed ""sensor""" --- p.15 / Chapter 1.5.1 --- ASN1 overexpressing lines (35S-ASN1) --- p.15 / Chapter 1.5.2 --- PII overexpressing lines (PII ox) & PII truncated lines (PII trunc.) --- p.16 / Chapter 1.5.2.1 --- Plant PII and its possible role --- p.16 / Chapter 1.5.2.2 --- Nitrogen/carbon sensing as the proposed in vivo function of PII-like protein in Arabidopsis thaliana by the in planta bioassay of PII overexpressing (PII ox) lines --- p.17 / Chapter 1.5.2.3 --- Changes in physiological and transcriptional expression of nitrogen assimilatory genes in PII transgenic lines --- p.17 / Chapter 1.6 --- Review on nitrogen controls and sensing mechanism of microbial organism and higher plants --- p.21 / Chapter 1.6.1 --- Nitrogen sensing in enteric bacteria --- p.21 / Chapter 1.6.2 --- Nitrogen sensing in cyanobacteria --- p.21 / Chapter 1.6.3 --- Nitrogen sensing in fungi --- p.22 / Chapter 1.6.4 --- Implication of the nitrogen sensing mechanisms in microorganisms to nitrogen sensing in plants --- p.23 / Chapter 1.7 --- "Hypothesis, objectives and outlines of this thesis work" --- p.25 / Chapter 2 --- Materials and Methods --- p.27-50 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- "Plants,bacterial strains and vectors" --- p.27 / Chapter 2.1.2 --- Chemicals and Regents --- p.28 / Chapter 2.1.3 --- "Buffer, solution and gel" --- p.28 / Chapter 2.1.4 --- Commercial kits --- p.28 / Chapter 2.1.5 --- Equipments and facilities used --- p.29 / Chapter 2.1.6 --- Growth medium --- p.29 / Chapter 2.1.7 --- Primers --- p.29 / Chapter 2.2 --- Methods --- p.29 / Chapter 2.2.1 --- Growth condition for plant materials --- p.29 / Chapter 2.2.1.1 --- General conditions --- p.29 / Chapter 2.2.1.2 --- Mature Arabidopsis for gene expression profile --- p.30 / Chapter 2.2.1.3 --- Arabidopsis seedlings for physiological experiment --- p.30 / Chapter 2.2.2 --- Molecular Techniques --- p.31 / Chapter 2.2.2.1 --- Bacterial cultures for recombinant DNA --- p.31 / Chapter 2.2.2.2 --- Preparation of pBluescript II KS(+) T-vector for cloning --- p.31 / Chapter 2.2.2.3 --- Cloning techniques --- p.32 / Chapter 2.2.2.4 --- Transformation of DH5a competent cell --- p.32 / Chapter 2.2.2.5 --- Gel electrophoresis --- p.33 / Chapter 2.2.2.6 --- DNA and RNA extractions from plant tissues --- p.34 / Chapter 2.2.2.7 --- First strand cDNA synthesis --- p.35 / Chapter 2.2.2.8 --- PCR techniques --- p.35 / Chapter 2.2.2.9 --- Sequencing --- p.37 / Chapter 2.2.3 --- Analysis of sequences and homology search --- p.37 / Chapter 2.2.4 --- Biochemical analysis --- p.41 / Chapter 2.2.4.1 --- Sugar content analysis --- p.41 / Chapter 2.2.4.2 --- Anthocyanin content analysis --- p.42 / Chapter 2.2.4.3 --- Fresh weight measurement --- p.43 / Chapter 2.2.4.4 --- Statistic analysis --- p.43 / Chapter 2.2.5 --- Generation of crossing progenies --- p.44 / Chapter 2.2.5.1 --- Artificial crossing of A. thaliana --- p.44 / Chapter 2.2.5.2 --- PCR screening for successful crossing --- p.44 / Chapter 2.2.6 --- Construction of subtractive libraries --- p.45 / Chapter 2.2.7 --- Reverse-dot blot screening --- p.45 / Chapter 2.2.7.1 --- in vitro transcription for making ampicillin cRNA --- p.46 / Chapter 2.2.7.2 --- PCR amplification --- p.47 / Chapter 2.2.7.3 --- Dot-blotting of PCR products on nylon membrane --- p.48 / Chapter 2.2.7.4 --- P probe preparation --- p.49 / Chapter 2.2.7.5 --- Hybridization --- p.49 / Chapter 2.2.7.6 --- Signal detection --- p.50 / Chapter 3 --- Results --- p.51-124 / Chapter 3.1 --- Differential growth behaviour and sugar content in 35S-ASNI lines --- p.51 / Chapter 3.1.1 --- Growth of the seedlings of 35S-ASN1 lines under different N and C supplementations --- p.51 / Chapter 3.1.2 --- Lowered reducing sugar content in 35S-ASN1 lines --- p.52 / Chapter 3.2 --- Development of markers for nitrogen signalling events related to altered N status in 35S-ASN1 lines --- p.60 / Chapter 3.2.1 --- Sugar-induced anthocyanin levels as common morphological marker shared by 35S-ASN1 lines and PII transgenic lines --- p.60 / Chapter 3.2.2 --- Expression markers related to altered N status in 35-ASN1 lines --- p.64 / Chapter 3.3 --- Generation of transgenic plants constitutively expressing ASN1 and GLB1 (or ASN1 and truncated GLB1) through crossing --- p.74 / Chapter 3.4 --- Search for homologs of well-known microbial nitrogen signalling components in Arabidopsis thaliana --- p.78 / Chapter 3.4.1 --- Homologs of yeast general amino acid control components --- p.80 / Chapter 3.4.1.1 --- Arabidopsis thaliana GCN2-like protein --- p.80 / Chapter 3.4.1.2 --- Arabidopsis thaliana GCN1 -like protein --- p.84 / Chapter 3.4.1.3 --- Arabidopsis thaliana GCN20-like protein --- p.84 / Chapter 3.4.1.4 --- Plant eIF2α --- p.86 / Chapter 3.4.1.5 --- Arabidopsis thaliana GCN4-CRE like sequences --- p.87 / Chapter 3.4.2 --- Homologs of fungi nitrogen sensing components: Globally acting factor in nitrogen control in fungi --- p.89 / Chapter 3.4.3 --- Homologs of cyanobacteria nitrogen control components: IF7 & IF 17 (Negative regulators of GS activity) --- p.89 / Chapter 4 --- Discussion --- p.125-147 / Chapter 4.1 --- Differential physiological and morphological behaviours found in the comparative studies between control lines and 35S-ASN1 lines --- p.125 / Chapter 4.1.1 --- in planta promotive effect of ASN1 overexpression on the seedlings growth under low nitrogen and in the absence of exogenous applied metabolizable sugar --- p.125 / Chapter 4.1.2 --- Modulation of sugar level in 35S-ASN1 lines --- p.126 / Chapter 4.2 --- Development of morphological marker and gene expression markers --- p.128 / Chapter 4.2.1 --- Anthocyanin accumulation as a morphological marker for epistatic analysis --- p.128 / Chapter 4.2.2 --- Differential expressed genes as candidates for gene expression markers of nitrogen signalling event --- p.131 / Chapter 4.3 --- Arabidopsis homolog search for well-known microbial signalling components --- p.132 / Chapter 4.3.1 --- "Possible amino acid sensing system in Arabidopsis constructed by homologs of yeast GCN2, GCN1, GCN20 and eIF2a" --- p.132 / Chapter 4.3.1.1 --- Arabidopsis GCN2-like (A. thaliana GCN2-like) protein --- p.132 / Chapter 4.3.1.2 --- Arabidopsis GCNl-like (A. thaliana GCNl-like) & GCN20-like (A. thaliana GCNl-like) proteins --- p.136 / Chapter 4.3.1.3 --- Plant eIF2a phosphorylation pathway --- p.139 / Chapter 4.3.1.4 --- GCN4 related transcriptional factors and GCN4-like motif (GLM) cis-element in plants --- p.140 / Chapter 4.3.1.5 --- Implication of the presence of plant homologs of fungi regulatory proteins involved in the general control of amino acid biosynthesis --- p.142 / Chapter 4.3.2 --- Failure in identifying homologs of nitrogen regulators responsible for switching of nitrogen source in Arabidopsis --- p.144 / Chapter 4.4 --- Overview of research platform construction --- p.146 / Chapter 5 --- Conclusion and Perspectives --- p.148 / Chapter 6 --- References --- p.149-155 / Chapter 7 --- Appendix --- p.156-167
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324454 |
| Date | January 2003 |
| Contributors | Chow, Cheung-ming., 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, xviii, 167 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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