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O arroz irrigado está entre os principais cereais produzidos no mundo. Determinar a evapotranspiração (ET) para as grandes áreas de arroz irrigado é um desafio devido a pouca disponibilidade de dados. Diversos Modelos de ET vêm sendo desenvolvidos com a intenção de monitorar áreas agrícolas, porém poucos estudos experimentais são realizados sobre áreas de arroz irrigado. Este trabalho tem como objetivo geral estimar a ET sobre cultivos de arroz irrigado através de um modelo que utiliza informações meteorológicas in situ (Priestley-Taylor) e outro remoto (MOD16). O Priestley-Taylor (PT) é um modelo de ET que utiliza informações de temperatura do ar e componentes relacionadas ao balanço de energia, juntamente com um parâmetro adimensional α. O modelo MOD16 foi criado para monitorar a ET em grandes áreas, utilizando informações meteorológicas de um banco de dados de reanálise juntamente com dados remotos. Ambos os métodos não apresentam calibração e validação sobre áreas de arroz irrigado no Sul do Brasil. Neste trabalho validamos estes dois modelos a partir de dois sítios experimentais com medidas de ET através da técnica de Eddy Covariance. Os resultados encontrados nesta pesquisa mostraram-se satisfatórios quando comparado o método PT com dados experimentais, recomendando-se a utilização de 1,22 do parâmetro α. A simplificação no método PT realizada a partir das componentes do balanço de energia, com substituição pela variável de radiação global através de uma regressão linear, mostrou-se válida apresentando erros poucos expressivos, e com valor de 1,18 para parâmetro α. Já o modelo de ET MOD16 mostrou-se pouco preciso sobre as áreas de arroz irrigado. A validação de ET MOD16 foi realizada sobre uma área de 3 x 3 km e pixel central, resultando em menor subestimativa do modelo para o pixel central em relação aos dados de Eddy Covariance. Além disso, a correlação foi realizada em função das variáveis ambientais, encontrando maior correlação do dado experimental com as componentes do balando de energia, enquanto o MOD16 apresentou maior correlação com a temperatura do ar. Por fim, sugerem-se melhorias na parametrização da energia disponível no modelo de ET MOD16. Além disso, a aplicação do método simplificado de PT é indicada sobre áreas de arroz irrigado. / Irrigated rice is among principal produced cereals in the world. Determining the evapotranspiration (ET) for large areas of irrigated rice is a challenge task, due to poor data availability. Several ET models have been developed with the intention of monitoring agricultural areas, however few experimental studies are accomplished on areas of irrigated rice. This study aims to estimate ET on irrigated rice crops using a model which employs meteorological information in situ (Priestley-Taylor) and one remote (MOD16). Priestley-Taylor (PT) is a model of ET that uses air temperature and related components to the energy balance as information and a dimensionless parameter α. The MOD16 model was designed to monitor ET in large areas using meteorological information obtained from the reanalysis database together with remote data. Both methods do not present calibration and validation on areas of irrigated rice in Southern Brazil. In this work we validate these two models from two experimental sites with ET measurements employing the Eddy Covariance technique. The results found in this research was satisfactory when compared to the PT method with experimental data. It was suggested 1.22 for the α parameter. The simplification in the PT method performed from the components of the energy balance, with substitution for the global radiation variable using a linear regression. It was validated with few expressive errors, with a value of 1.18 for α parameter. On the other hand, the MOD16 model did not showed good accuracy on the areas of irrigated rice. The validation of ET MOD16 was performed over an area of 3 x 3 km and central pixel, resulting in a small underestimation of the model for the central pixel in relation to Eddy Covariance data. In addition, was performed the correlation in function of the environmental variables, finding a higher correlation of the experimental data with the components of the energy balance, while the MOD16 showed a high correlation with the air temperature. Finally, it was suggested improvements in the parameterization of the available energy in the model of ET MOD16, and to indicate the application of the simplified method of PT on the areas with irrigated rice.

Ma Junhao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 111-123). / Abstracts in English and Chinese. / Thesis Committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgements --- p.vii / General Abbreviations --- p.ix / Abbreviations of Chemicals --- p.xi / Table of Contents --- p.xii / List of Figures --- p.xvi / List of Tables --- p.xix / Chapter 1. --- Literature review --- p.1 / Chapter 1.1 --- Importance of amino acid metabolism --- p.1 / Chapter 1.1.1 --- Rice as the important source of essential amino acids --- p.1 / Chapter 1.1.2 --- Rice seeds are nutritionally incomplete --- p.2 / Chapter 1.1.3 --- The nitrogen source for aspartate family amino acid synthesis --- p.3 / Chapter 1.1.4 --- Synthesis of aspartate family amino acids in plants --- p.4 / Chapter 1.1.5 --- Regulation of the aspartate pathway at free amino acid level --- p.12 / Chapter 1.2 --- Regulation of amino acid metabolism during seed development --- p.14 / Chapter 1.3 --- Signalling system for nitrogen metabolism --- p.17 / Chapter 1.3.1 --- Nitrogen signalling in plants --- p.18 / Chapter 1.3.2 --- GCN signalling: another nitrogen signalling pathway --- p.21 / Chapter 1.3.2.1 --- Mechanism of GCN signalling pathway in yeast --- p.21 / Chapter 1.3.2.2 --- GCN in mammalian --- p.24 / Chapter 1.3.2.3 --- GCN in higher plant --- p.24 / Chapter 1.3.3 --- Relationship between carbon and nitrogen metabolic signaling in plants --- p.26 / Chapter 1.3.4 --- Paradigm for elucidating new signal transduction pathways --- p.29 / Chapter 1.4 --- Hypothesis of this thesis work --- p.30 / Chapter 2. --- Materials and Methods --- p.32 / Chapter 2.1. --- Materials --- p.32 / Chapter 2.1.1 --- Plants --- p.32 / Chapter 2.1.2 --- Bacterial strains and vectors --- p.32 / Chapter 2.1.3 --- Chemicals and reagents --- p.33 / Chapter 2.1.4 --- Buffer，solution and gel --- p.33 / Chapter 2.1.5 --- Commercial kits --- p.33 / Chapter 2.1.6 --- Equipments and facilities used --- p.33 / Chapter 2.1.6 --- Growth medium --- p.34 / Chapter 2.2 --- Methods --- p.34 / Chapter 2.2.1 --- Profiling genes expression pattern in developing rice seeds --- p.34 / Chapter 2.2.1.1 --- Growth conditions of rice --- p.34 / Chapter 2.2.1.2 --- Collection of developing rice seeds --- p.35 / Chapter 2.2.1.3 --- Total RNA extraction from rice seeds --- p.37 / Chapter 2.2.1.4 --- Total RNA extraction from plant leaf --- p.37 / Chapter 2.2.1.5 --- Gel electrophoresis --- p.38 / Chapter 2.2.1.6 --- First strand cDNA synthesis from rice total RNA --- p.39 / Chapter 2.2.1.7 --- Search for the coding sequence of rice genes related to amino acid metabolism --- p.39 / Chapter 2.2.1.8 --- Alignment of homologous coding sequence between family member genes --- p.42 / Chapter 2.2.1.9 --- Primer design --- p.42 / Chapter 2.2.1.10 --- Quantitation of total RNA and determination of internal control --- p.45 / Chapter 2.2.1.11 --- PCR to amplify the DNA fragments --- p.45 / Chapter 2.2.1.12 --- DNA Sequencing --- p.46 / Chapter 2.2.1.13 --- Generation and testing of single-stranded DIG-labelled DNA probes --- p.46 / Chapter 2.2.1.14 --- Northern blot --- p.47 / Chapter 2.2.1.15 --- RT-PCR (Reverse-transcription polymerase chain reaction) --- p.48 / Chapter 2.2.2 --- Expression assay of selected genes in herbicide treated plants --- p.49 / Chapter 2.2.2.1 --- Growing conditions and herbicide treatments --- p.49 / Chapter 2.2.2.2 --- GCN2 homologue in Arabidopsis and rice --- p.51 / Chapter 2.2.2.3 --- RT-PCR to analyze the change in expression level of selected genes in herbicide treated plants --- p.53 / Chapter 2.2.3 --- Generation of transgenic Arabidopsis --- p.56 / Chapter 2.2.3.1 --- Preparation of T-vector for T-ligation --- p.56 / Chapter 2.2.3.2 --- Cloning of Arabidopsis GCN2 gene --- p.56 / Chapter 2.2.3.3 --- Transformation of the plasmid into DH5a competent cell --- p.57 / Chapter 2.2.3.4 --- Screening of right recombinants --- p.58 / Chapter 2.2.3.5 --- Construction of chimeric AtGCN2 genes --- p.59 / Chapter 2.2.3.6 --- Transformation of electro-competent Agrobacterium cell --- p.60 / Chapter 2.2.3.7 --- Transformation of Arabidopsis by vacuum infiltration --- p.61 / Chapter 2.2.3.8 --- Selection of hemizygous and homozygous transgenic plants --- p.61 / Chapter 2.2.3.9 --- Screening of the T3 transformants --- p.63 / Chapter 2.2.3.10 --- Expression analysis of homozygous AtGCN2 transgenic Arabidopsis --- p.63 / Chapter 3. --- Results --- p.63 / Chapter 3.1 --- Profiling genes expression pattern in developing rice seeds --- p.63 / Chapter 3.1.1 --- Quantification of total RNA from seeds at different developing stages --- p.63 / Chapter 3.1.2 --- DNA sequence analysis --- p.66 / Chapter 3.1.3 --- Profiling the gene expression in developing rice seeds --- p.67 / Chapter 3.1.3.1 --- Expression profiles of nitrogen assimilation related genes --- p.67 / Chapter 3.1.3.2 --- Expression profiles of aspartate pathway genes --- p.72 / Chapter 3.1.3.3 --- Expression profiles of branched-chain amino acid synthesis pathway genes --- p.78 / Chapter 3.2 --- Relationship between GCN2 and amino acid metabolism in plants --- p.82 / Chapter 3.2.1 --- GCN2 homologue in A. thaliana and rice --- p.82 / Chapter 3.2.2 --- GCN2 and amino acid starvation --- p.85 / Chapter 3.2.3 --- Effects of amino acid starvation on GCN2 expression --- p.90 / Chapter 3.2.3 --- Changes in the expression level AK and BCAT genes in herbicide treated rice and A. thaliana --- p.93 / Chapter 3.3 --- Characterization of GCN2 transgenic A. thaliana --- p.96 / Chapter 3.3.1 --- Construct of pBI121-AtGCN2 --- p.96 / Chapter 3.3.2 --- Construction of GCN2 transgenic A. thaliana --- p.96 / Chapter 3.3.3 --- Expression of GCN2 in transgenic A. thaliana --- p.97 / Chapter 3.3.4 --- Expression level changes of AK and BCAT in transgenic A. thaliana --- p.99 / Chapter 4. --- Discussions --- p.101 / Chapter 4.1 --- Expression pattern of selected metabolic genes in developing plant seeds --- p.101 / Chapter 4.1.1 --- Most genes studied displayed a similar pattern --- p.101 / Chapter 4.1.2 --- Regulation of gene expression in developing rice seeds --- p.105 / Chapter 4.2 --- GCN2 and its role in higher plants --- p.106 / Chapter 4.2.1 --- The existence of the GCN2 gene in rice --- p.106 / Chapter 4.2.2 --- GCN2 responses solely to amino acid starvation --- p.106 / Chapter 5. --- Conclusion and Prospective --- p.109 / Reference --- p.111 / Appendix I: Chemicals and reagents --- p.124 / "Appendix II: Buffer, solution and gel" --- p.126 / Appendix III: Commercial kits --- p.128 / Appendix IV: Equipments and facilities used --- p.128

Dong Biao. / "October 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 184-201) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

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Yu, Wai Han. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 87-101). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.iii / ABSTRACT --- p.iv / 摘要 --- p.vi / LIST OF CONTENTS --- p.viii / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xiv / LIST OF ABBREVIATIONS --- p.xv / Chapter CHAPTER 1. --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.3 / Chapter 2.1 --- The importance of rice --- p.3 / Chapter 2.2 --- Limitation of essential amino acids in rice --- p.4 / Chapter 2.3 --- Lysine biosynthetic pathway --- p.6 / Chapter 2.3.1 --- The biosynthesis of aspartate --- p.6 / Chapter 2.3.2 --- Aspartate family pathway --- p.3 / Chapter 2.3.2.1 --- Aspartate kinase (AK) --- p.10 / Chapter 2.3.2.2 --- Dihydrodipicolinate synthase (DHPS) --- p.12 / Chapter 2.3.2.3 --- Other enzymes --- p.14 / Chapter 2.4 --- Regulation of lysine content in plant --- p.15 / Chapter 2.5 --- Enhancement of lysine content in plants --- p.16 / Chapter 2.5.1 --- "Breeding, selection and naturally occuring muatnts" --- p.17 / Chapter 2.5.2 --- Induced biochemical mutants --- p.18 / Chapter 2.5.3 --- Transgenic plants --- p.19 / Chapter 2.6 --- Hypothesis --- p.24 / Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.25 / Chapter 3.1 --- Introduction --- p.25 / Chapter 3.2 --- Chemicals --- p.25 / Chapter 3.3 --- Bacterial strains --- p.25 / Chapter 3.4 --- Cloning of AK and DHPS cDNAs --- p.25 / Chapter 3.4.1 --- Plant materials --- p.25 / Chapter 3.4.2 --- RNA extraction --- p.26 / Chapter 3.4.3 --- RT-PCR amplification of AK and DHPS cDNAs --- p.26 / Chapter 3.4.4 --- Sequence modification of AK and DHPS cDNAs --- p.27 / Chapter 3.4.5 --- DNA sequencing of AK and DHPS cDNAs --- p.32 / Chapter 3.5 --- Chimeric gene construction for rice transformation --- p.32 / Chapter 3.5.1 --- Plasmid and genetic material --- p.32 / Chapter 3.5.2 --- Construction of chimeric genes with seed-specific promoter --- p.35 / Chapter 3.5.3 --- Sequence fidelity of chimeric genes --- p.37 / Chapter 3.6 --- AEC resistance of E.coli expressing modified AK and DHPS --- p.37 / Chapter 3.7 --- Rice transformation --- p.38 / Chapter 3.7.1 --- Plant materials --- p.38 / Chapter 3.7.2 --- Preparation of agrobacterium --- p.33 / Chapter 3.7.3 --- Agrobacterium-mediated rice transformation --- p.39 / Chapter 3.7.3.1 --- Callus induction from mature rice seed embryos --- p.39 / Chapter 7.3.2 --- "Co-cultivation, selection and regeneration of transgenic rice" --- p.39 / Chapter 3.8 --- Analysis of transgenic expression --- p.41 / Chapter 3.8.1 --- Genomic DNA extraction --- p.41 / Chapter 3.8.2 --- Total RNA extraction --- p.41 / Chapter 3.8.3 --- Synthesis of DIG-labeled DNA probe --- p.42 / Chapter 3.8.4 --- Southern blot analysis --- p.43 / Chapter 3.8.5 --- Northern blot analysis --- p.43 / Chapter 3.8.6 --- Extraction of rice seed protein --- p.43 / Chapter 3.8.7 --- Tricine SDS-PAGE --- p.44 / Chapter 3.8.8 --- Raising AK and DHPS antibody --- p.44 / Chapter 3.8.9 --- Western blot analysis --- p.46 / Chapter 3.9 --- Free amino acid analysis --- p.46 / Chapter CHAPTER 4. --- RESULTS --- p.48 / Chapter 4.1 --- Cloning of AK and DHPS cDNAs from rice --- p.48 / Chapter 4.1.1 --- RNA extraction and cDNAs amplification --- p.43 / Chapter 4.1.2 --- Sequencing of AK and DHPS cDNAs --- p.50 / Chapter 4.2 --- Sequence modification of AK and DHPS cDNAs --- p.50 / Chapter 4.3 --- Construction of chimeric genes --- p.50 / Chapter 4.4 --- AEC resistance of E.coli expressing modified AK and DHPS --- p.56 / Chapter 4.5 --- Rice transformation --- p.58 / Chapter 4.6 --- Detection of target genes in transgenic rice lines --- p.60 / Chapter 4.6.1 --- PCR of genomic DNA --- p.60 / Chapter 4.6.2 --- Southern blot analysis --- p.63 / Chapter 4.7 --- Northern blot analysis --- p.65 / Chapter 4.8 --- Western blot analysis of AK and DHPS proteins --- p.66 / Chapter 4.9 --- Free amino acid analysis --- p.68 / Chapter 4.9.1 --- Free lysine content --- p.68 / Chapter 4.9.2 --- Changes in other amino acids --- p.69 / Chapter CHAPTER 5. --- DISCUSSION --- p.82 / Chapter 5.1 --- Cloning and modification of AK and DHPS cDNAs --- p.82 / Chapter 5.2 --- Seed-specific expression of modified AK and DHPS in rice --- p.82 / Chapter 5.3 --- Free amino acid changes in transgenic rice lines --- p.83 / Chapter 5.4 --- Future perspectives --- p.85 / Chapter CHAPTER 6. --- CONCLUSION --- p.86 / REFERENCES --- p.87 / APPENDIX --- p.102

Leung, Chiu Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 219-235). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.iii / ABSTRACT --- p.v / 摘要 --- p.vii / TABLE OF CONTENTS --- p.ix / LIST OF FIGURES --- p.xiv / LIST OF TABLES --- p.xviii / LIST OF ABBREVIATIONS --- p.xix / Chapter CHAPTER 1. --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.5 / Chapter 2.1 --- Introduction to carotenoids --- p.5 / Chapter 2.1.1 --- Structures and general chemical properties --- p.6 / Chapter 2.1.2 --- Dietary source of carotenoids --- p.8 / Chapter 2.2 --- Biosynthesis of carotenoids in plants --- p.8 / Chapter 2.2.1 --- Formation of isopretenyl diphosphate (IPP) --- p.12 / Chapter 2.2.2 --- "Formation of C40 backbone, phytoene" --- p.13 / Chapter 2.2.3 --- Desaturation reactions --- p.16 / Chapter 2.2.4 --- Isomerization --- p.18 / Chapter 2.2.5 --- Cyclization --- p.18 / Chapter 2.2.6 --- Xanthophylls synthesis --- p.19 / Chapter 2.2.6.1 --- Hydroxylation --- p.19 / Chapter 2.2.6.2 --- Epoxidation and de-epoxidation --- p.19 / Chapter 2.2.6.3 --- Neoxanthin formation --- p.20 / Chapter 2.2.7 --- Carotenoids catabolism by cleavage enzymes --- p.20 / Chapter 2.2.8 --- Carotenoids sequestration --- p.20 / Chapter 2.2.9 --- Regulations of Carotenogenesis in plant --- p.21 / Chapter 2.3 --- Roles of carotenoids in plants --- p.24 / Chapter 2.3.1 --- Precursor of abscisic acid (ABA) production --- p.24 / Chapter 2.3.2 --- Photomorphogenesis: Prolamella body formation --- p.27 / Chapter 2.3.3 --- "Light-harvesting, energy transfer and photoprotection" --- p.30 / Chapter 2.4 --- Importance of carotenoids to human --- p.34 / Chapter 2.4.1 --- Provitamin A activity and conversion --- p.34 / Chapter 2.4.2 --- Roles of vitamin A and carotenoids in diseases prevention --- p.36 / Chapter 2.4.2.1 --- Visual cycle and related diseases --- p.36 / Chapter 2.4.2.2 --- Cardiovascular diseases --- p.37 / Chapter 2.4.2.3 --- Cancer --- p.38 / Chapter 2.4.3 --- Roles of vitamin A in gene regulation --- p.39 / Chapter 2.4.4 --- Bioavailability and daily intake recommendation --- p.39 / Chapter 2.5 --- Vitamin A deficiency (VAD) --- p.42 / Chapter 2.5.1 --- Clinicopathological features --- p.44 / Chapter 2.5.1.1 --- Visual problems --- p.44 / Chapter 2.5.1.2 --- Infection --- p.45 / Chapter 2.6 --- Global efforts in combating VAD --- p.45 / Chapter 2.6.1 --- Dietary diversification --- p.46 / Chapter 2.6.2 --- Supplementation --- p.48 / Chapter 2.6.3 --- Pro-vitamin A enriched crops by genetical engineering --- p.49 / Chapter 2.6.3.1 --- Tomato --- p.49 / Chapter 2.6.3.2 --- Potato --- p.50 / Chapter 2.6.3.3 --- Canola --- p.50 / Chapter 2.6.3.4 --- The Golden Rice (GR) project --- p.51 / Chapter 2.6.3.4.1 --- The 1st generation (GR1) --- p.52 / Chapter 2.6.3.4.2 --- The 2nd generation (GR2) --- p.54 / Chapter 2.7 --- Rice phytoene synthase as a GR candidate enzyme --- p.55 / Chapter 2.7.1 --- General properties of phytoene synthase in higher plant --- p.55 / Chapter 2.7.1.1 --- Gene duplication and structure --- p.55 / Chapter 2.7.1.2 --- Membrane association and cation requirement --- p.57 / Chapter 2.7.1.3 --- Expression pattern and tissue-specificity --- p.58 / Chapter 2.7.2 --- "Rice phytoene synthases: Ospsy1, Ospsy2 and Ospsy3" --- p.60 / Chapter 2.7.2.1 --- Gene duplication and structure --- p.60 / Chapter 2.7.2.2 --- Protein structures and phylogenetic analysis --- p.60 / Chapter 2.7.2.3 --- Expression pattern --- p.62 / Chapter 2.7.2.4 --- Light- and stress-induced expression --- p.63 / Chapter 2.7.3 --- Rice phytoene synthases activity in rice seeds --- p.64 / Chapter 2.7.3.1 --- Previous study in rice seed carotenogenic capacity --- p.64 / Chapter 2.8 --- Seed-specific rice promoters --- p.66 / Chapter 2.8.1 --- Previous studies on seed-specific expression in rice --- p.66 / Chapter 2.8.1.1 --- Endosperm-specific cis-acting regulatory elements --- p.67 / Chapter 2.8.1.2 --- Requirements to confer endosperm-specific expression in rice --- p.69 / Chapter 2.9 --- Project overview and hypothesis --- p.71 / Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.73 / Chapter 3.1 --- Chemicals --- p.73 / Chapter 3.2 --- Vectors and bacterial strains in regular cloning --- p.73 / Chapter 3.3 --- Plant materials --- p.74 / Chapter 3.4 --- Construction of gene cassettes for plant transformation --- p.74 / Chapter 3.4.1 --- Construction of gene cassettes for Ospsyl and Ospsy2 study --- p.74 / Chapter 3.4.1.1 --- Cloning of Ospsyl and Ospsy2 from rice --- p.76 / Chapter 4.1.1.1 --- Total RNA extraction --- p.75 / Chapter 3.4.1.1.2 --- Amplification of cDNA of Ospsyl by RT-PCR --- p.77 / Chapter 3.4.1.1.3 --- Amplification of cDNA of Ospsy2 by PCR --- p.80 / Chapter 3.4.1.2 --- Cloning of constitutive CaMV35S promoter --- p.82 / Chapter 3.4.1.2.1 --- Preparation of pBI121 vector --- p.82 / Chapter 3.4.1.2.2 --- Amplification of CaMV35S promoter by PCR --- p.82 / Chapter 3.4.1.3 --- Cloning of endosperm-specific rice glutelin-1 (Gt-1) promoter --- p.84 / Chapter 3. 4.1.3.1 --- Genomic DNA extraction --- p.84 / Chapter 3.4.1.3.2 --- Amplification of Gt-1 promoter by PCR --- p.84 / Chapter 3.4.1.4 --- Construction of gene cassettes for Ospsyl and Ospsy2 driven by CaMV35S and rice Gt-1 promoter --- p.87 / Chapter 3.4.2 --- Construction of gene cassettes for promoter analysis --- p.90 / Chapter 3.4.2.1 --- Cloning of full length and fragments of Ospsyl promoter --- p.92 / Chapter 3.4.2.1.1 --- Genomic DNA extraction --- p.95 / Chapter 3.4.2.1.2 --- Amplification of full length and fragments of Ospsy1 promoter --- p.95 / Chapter 3.4.2.2 --- Cloning of CaMV35S minimal promoter --- p.96 / Chapter 3.4.2.2.1 --- Amplification of CaMV35S minimal promoter --- p.97 / Chapter 3.4.2.3 --- Cloning of rice Gt-1 promoter --- p.97 / Chapter 3.4.2.3.1 --- Genomic DNA extraction --- p.97 / Chapter 3.4.2.3.2 --- Amplification of Gt-1 promoter --- p.97 / Chapter 3.4.2.4 --- Annealing of linker --- p.93 / Chapter 3.4.3.5 --- Making of rice GCN4 multimers --- p.98 / Chapter 3.4.2.6 --- Construction of gene cassettes for promoter analysis --- p.99 / Chapter 3.4.2.7 --- Construction of gene cassettes for Gt-1 promoter analysis --- p.104 / Chapter 3.4.3 --- Confirmation of sequence fidelity --- p.106 / Chapter 3.5 --- Rice transformation --- p.107 / Chapter 3.5.1 --- Plant materials --- p.107 / Chapter 3.5.2 --- Preparation of Agrobacterium --- p.107 / Chapter 3.5.3 --- Agrobacterium mediated transformation --- p.108 / Chapter 3.5.4 --- Callus induction from mature rice seeds --- p.109 / Chapter 3.5.6 --- Co-cultivation and selection --- p.109 / Chapter 3.5.7 --- Pre-regeneration and regeneration of transgenic rice --- p.110 / Chapter 3.5.8 --- Plantation of transgenic rice --- p.110 / Chapter 3.6 --- Detection of transgene expression --- p.112 / Chapter 3.6.1 --- Detection at DNA level --- p.112 / Chapter 3.6.1.1 --- Genomic DNA extraction --- p.112 / Chapter 3.6.1.2 --- PCR screening --- p.122 / Chapter 3.6.1.3 --- Synthesis of DIG-labeled DNA probes --- p.116 / Chapter 3.6.1.4 --- Southern blot analysis --- p.118 / Chapter 3.6.2 --- Detection at RNA level --- p.119 / Chapter 3.6.2.1 --- Total RNA extraction --- p.119 / Chapter 3.6.2.2 --- Northern blot analysis --- p.119 / Chapter 3.6.3 --- Detection at protein level --- p.119 / Chapter 3.6.3.1 --- Antibody production --- p.119 / Chapter 3.6.3.1 --- Ospsyl and Ospsy2 induction in pET system --- p.120 / Chapter 3.63.1.2 --- Immunization of rabbit and serum collection --- p.123 / Chapter 3.6.3.2 --- Total protein extraction from plant materials --- p.124 / Chapter 3.6.3.2.1 --- Protein extraction from rice calli and leaves --- p.124 / Chapter 3.6.3.2.2 --- Protein extraction from immature and mature rice seeds --- p.124 / Chapter 3.6.3.3 --- Tricine SDS-PAGE --- p.125 / Chapter 3.6.3.4 --- Western blot analysis --- p.125 / Chapter 3.6.4 --- Detection at metabolite level --- p.126 / Chapter 3.6.4.1 --- Isoprenoids extraction from plant materials --- p.126 / Chapter 3.6.4.2 --- UPLC analysis for isoprenoid identification --- p.127 / Chapter 3.6.5 --- Detection of promoter activity --- p.128 / Chapter 3.6.5.1 --- Histochemical staining of GUS activity --- p.128 / Chapter 3.6.5.1.1 --- Histochemical staining --- p.128 / Chapter 3.6.5.1.2 --- Plant tissue fixation for microscopic observation --- p.128 / Chapter 3.6.5.2 --- GUS activity assay --- p.129 / Chapter 3.6.5.2.1 --- Protein extraction and quantitation with Bio-Rad protein assay --- p.129 / Chapter 3.6.5.2.2 --- G US activity assay --- p.130 / Chapter CHAPTER 4. --- RESULTS --- p.131 / Chapter 4.1 --- Tissue-specificity and endosperm specific expression of rice psy1 and Psy2 --- p.131 / Chapter 4.1.1 --- Construction of gene cassettes for study in Ospsy1 and Ospsy2 --- p.133 / Chapter 4.1.2 --- Rice transformation --- p.135 / Chapter 4.1.3 --- Transgene detection --- p.137 / Chapter 4.1.3.1 --- Genomic DNA PCR screening --- p.137 / Chapter 4.1.3.2 --- Southern blot analysis --- p.139 / Chapter 4.1.3.2.1 --- Southern blot analysis on transgenic rice calli --- p.141 / Chapter 4.1.3.2.2 --- Southern blot analysis on regenerated rice --- p.143 / Chapter 4.1.4 --- Detection of transgene expression --- p.149 / Chapter 4.1.4.1 --- Northern blot analysis on immature transgenic seed --- p.149 / Chapter 4.1.4.2 --- Western blot analysis on transgenic rice tissues --- p.152 / Chapter 4.1.4.2.1 --- Antibody production --- p.152 / Chapter 4.1.4.2.2 --- Western blot analysis of transgenic rice calli --- p.155 / Chapter 4.1.4.2.3 --- Western blot analysis of transgenic rice leaves --- p.157 / Chapter 4.1.4.2.4 --- Western blot analysis of immature transgenic rice seeds --- p.160 / Chapter 4.1.5 --- Detection of OsPSYs activity at metabolite level --- p.163 / Chapter 4.1.5.1 --- UPLC analysis on transgenic rice tissues --- p.163 / Chapter 4.1.5.1.1 --- Carotenoid profiling of transgenic rice calli --- p.163 / Chapter 4.1.5.1.2 --- Carotenoid profiling of transgenic rice leaves --- p.168 / Chapter "4.1.5.1,3" --- Carotenoid profiling of mature transgenic rice seeds --- p.172 / Chapter 4.2 --- Promoter analysis of modified rice psy1 promoter --- p.176 / Chapter 4.2.1 --- Construction of gene cassettes for promoter analysis --- p.178 / Chapter 4.2.2 --- Rice transformation --- p.180 / Chapter 4.2.3 --- Transgene detection --- p.180 / Chapter 4.2.3.1 --- Genomic DNA PCR screening --- p.180 / Chapter 4.2.3.2 --- Southern blot analysis --- p.185 / Chapter 4.2.3.2.1 --- Southern blot analysis of regenerated rice --- p.186 / Chapter 4.2.4 --- Detection of promoter activity --- p.196 / Chapter 4.2.4.1 --- Promoter activity in transgenic rice leaves --- p.196 / Chapter 4.2.4.1.1 --- Histochemical staining of GUS --- p.196 / Chapter 4.2.4.1.2 --- G US activity assay --- p.199 / Chapter 4.2.4.2 --- Promoter activity in transgenic immature seeds --- p.202 / Chapter 4.2.4.2.1 --- Histochemical staining of GUS --- p.202 / Chapter 4.2.4.2.2 --- GUS activity assay --- p.206 / Chapter CHAPTER 5. --- DISCUSSIONS --- p.209 / Chapter 5.1 --- Tissue-specificity and endosperm specific expression of rice psyl and psy2 --- p.209 / Chapter 5.1.1 --- OsPSYl and OsPSY2 activities in rice calli --- p.209 / Chapter 5.1.2 --- OsPSYl and OsPSY2 activities in rice leaves --- p.210 / Chapter 5.1.3 --- OsPSYl and OsPSY2 activities in rice seeds --- p.211 / Chapter 5.2 --- Analysis of modified rice psyl promoter --- p.213 / Chapter 5.3 --- Future prospects of Golden Rice --- p.214 / Chapter CHAPTER 6. --- CONCLUSIONS --- p.217 / REFERENCES --- p.219

Liu, Yang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 99-104). / Abstracts in English and Chinese. / Table of Contents / Thesis/Assessment Committee --- p.ii / Statement --- p.iii / Abstract --- p.iv / 摘要 --- p.vi / Acknowledgements --- p.vii / Table of Contents --- p.viii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1. --- The plant secretory pathway --- p.2 / Chapter 2. --- Vesicular pathways in plant cells --- p.3 / Chapter 3. --- Prevacuolar Compartments --- p.6 / Chapter 4. --- Vacuolar sorting receptors (VSRs) --- p.7 / Chapter 5. --- BP-80 & Arabidopsis VSR Proteins --- p.8 / Chapter 6. --- Research Objectives --- p.9 / Chapter Chapter 2 --- Development and Expression of GFP-OsVSRs Fusion Reporters in Tobacco BY-2 and Rice Suspension Cultured Cells --- p.11 / Chapter 1. --- Introduction --- p.12 / Chapter 2. --- Materials and methods --- p.14 / Chapter 2.1 --- Construction of GFP-OsVSR chimeric reporters --- p.14 / Chapter 2.2 --- Construction of Golgi Marker and PVC marker --- p.18 / Chapter 2.3 --- Agrobacterium electroporation --- p.27 / Chapter 2.4 --- Transformation of tobacco BY-2 cells --- p.27 / Chapter 2.5 --- Transient expression of GFP-OsVSRs in protoplasts of tobacco BY-2 cells and rice suspension cultured cells --- p.28 / Chapter 2.6 --- Screening of transgenic BY-2 cells expressing GFP-OsVSR reporters --- p.30 / Chapter 2.7 --- Chemicals --- p.33 / Chapter 3. --- Results --- p.34 / Chapter 3.1 --- Study subcellular localization of OsVSR proteins with chimeric GFP-OsVSR reporters --- p.34 / Chapter 3.2 --- Generation of transgenic tobacco BY-2 cell lines expressing GFP-OsVSR reporter constructs --- p.38 / Chapter 3.3 --- Transient expression of GFP-OsVSR reporters in tobacco BY-2 and rice cell protoplasts --- p.40 / Chapter 4. --- Conclusion --- p.42 / Chapter Chapter 3 --- Subcellular Localization of GFP-OsVSR Fusion Reporters in Tobacco BY-2 and Rice Suspension Cultured Cells --- p.43 / Chapter 1. --- Introduction --- p.44 / Chapter 2. --- Materials and methods --- p.45 / Chapter 2.1 --- Confocal immunofluorescence studies --- p.45 / Chapter 2.2 --- Antibodies --- p.46 / Chapter 2.3 --- Wortmannin and BFA drug treatment --- p.46 / Chapter 2.4 --- Electron microscopy of resin-embedded cells --- p.47 / Chapter 2.5 --- Two-dimensional (2-D) gel analysis --- p.47 / Chapter 3 --- Results --- p.49 / Chapter 3.1 --- "Distinct subcellular localizations of GFP-OsVSRl, GFP-OsVSR2 and GFP-OsVSR4 reporters in transgenic BY-2 cell lines" --- p.49 / Chapter 3.2 --- "Subcellular localizations of GFP-OsVSRl, GFP-OsVSR2 and GFP-OsVSR4 in protoplasts of rice suspension cultured cells" --- p.58 / Chapter 3.3 --- Distinct localizations of GFP-OsVSR3 and GFP-OsVSR5 --- p.62 / Chapter 3.4 --- Immunogold EM localization of VSR proteins in rice suspension cultured cells --- p.65 / Chapter 3.5 --- 2-D western blot detection of VSR proteins in various plants --- p.68 / Chapter 4. --- Conclusions --- p.70 / Chapter Chapter 4 --- Summary and Discussion --- p.71 / Chapter 1. --- The significance of this study --- p.72 / Chapter 2. --- The hypothesis in this study --- p.73 / Chapter 3. --- A reporter system to study subcellular localization of OsVSR proteins in both tobacco BY-2 cells and rice suspension cultured cells --- p.75 / Chapter 4. --- Transiently expression of GFP-OsVSR reporters in BY-2 and rice protoplasts ..… --- p.76 / Chapter 5. --- Distinct PVC and Golgi localizations of GFP-OsVSR fusions --- p.77 / Chapter 6. --- Summary and future perspective --- p.78 / Appendix --- p.79 / Characterization of A Novel Rice Protein --- p.79 / Chapter 1. --- Introduction --- p.80 / Chapter 2. --- Materials and methods --- p.83 / Chapter 2.1 --- Antibodies --- p.83 / Chapter 2.2 --- Sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) and western blot analysis of proteins from different plant species --- p.84 / Chapter 2.3 --- Sucrose gradient fractionation with protein F antibody --- p.84 / Chapter 2.4 --- Confocal immunofluorescence studies --- p.85 / Chapter 2.5 --- Affinity purification of Protein F by its antibody --- p.85 / Chapter 3. --- Results --- p.87 / Chapter 3.1 --- Protein F is presented in different plant species --- p.87 / Chapter 3.2 --- Protein F is an integral membrane protein --- p.89 / Chapter 3.3 --- Subcelluar localization of Protein F --- p.91 / Chapter 3.4 --- Affinity purification of Protein F for identification --- p.95 / Chapter 4. --- Summary and future perspectives --- p.97 / Chapter 4.1 --- Summary --- p.97 / Chapter 4.2 --- Future Perspectives --- p.97 / References --- p.99

Excerpt: The good folks at the Mountains Of Music Homecoming who bring 9 days of music, culture, and culinary events to southwestern Virginia each June, have begun to publish short interviews with the various personalities associated with these ventures...Such includes this interview with Wyatt Rice recently conducted by Ted Olson, professor of Appalachian Studies at East Tennessee State University. Wyatt is the younger brother of the legendary Tony Rice, and performed alongside him for most of his career.

The current atmospheric CO2 partial pressure of 36 Pa is expected to nearly double by the end of the 21st Century.Increases of this magnitude are likely to profoundly change the biochemistry, growth and morphology of plants, particularly C3 species.The research in this thesis focuses on the micronutrient Zinc (Zn), because this element is associated with a number of macromolecules which play key roles in plant growth and development, particularly on the shoot apex.The main objective of the work was to study the influence of elevated CO2 Zn nutrition of rice in the vegetative phase.A second objective was to investigate whether high CO2 reduced Zn concentrations in grain of cv. Jarrah and a Japanese cultivar, Akitakomachi, grown in either controlled environments, or in the field in a FACE (Free Air CO2 Enrichment) experiment. The greater Zn use efficiency of cvv. IR8 and Jarrah at elevated CO2, and the fact that high CO2 completely overcame chronic Zn deficiency at low Zn supplies, indicates that it may be possible , under future CO2 scenarios, to produce rice in areas where low soil Zn availability currently limits yield. / Master of Science (Hons)