| 摘要 | 第6-7页 |
| Abstract | 第7-8页 |
| Chapter 1. Introduction | 第18-48页 |
| 1.1 Sugarcane | 第18-20页 |
| 1.1.1 Brief history of sugarcane crop | 第18-19页 |
| 1.1.2 Classification of sugarcane | 第19-20页 |
| 1.2 Genetic engineering of sugarcane | 第20-25页 |
| 1.2.1 Biolistics Transformation | 第21-23页 |
| 1.2.2 Agrobacterim-mediated Transformation | 第23-25页 |
| 1.3 Stress Sensing and Signal Transduction | 第25-26页 |
| 1.4 Regulation of gene expression | 第26-27页 |
| 1.4.1 Gene regulation at multiple levels | 第26-27页 |
| 1.4.2 Regulation at transcriptional level | 第27页 |
| 1.5 Ethylene responsive factors (ERF) Gene Family | 第27-33页 |
| 1.5.1 Classification of ERFs | 第28-29页 |
| 1.5.2 Role of ERF members in abiotic stress response | 第29-33页 |
| 1.6 Effects of abiotic stresses on plants | 第33-35页 |
| 1.6.1 Plant Response Mechanisms to Abiotic Stresses | 第33-35页 |
| 1.7 Drought stress | 第35-39页 |
| 1.7.1 Plant Responses to Drought Stress | 第36-38页 |
| 1.7.2 Molecular mechanisms activate during drought | 第38-39页 |
| 1.8 Salt stress | 第39-43页 |
| 1.8.1 Physiological responses of plants to effects of salt stress | 第40-43页 |
| 1.9 Oxidative Stress | 第43-44页 |
| 1.9.1 Reactive oxygen species | 第43-44页 |
| 1.9.2 ROS scavenging | 第44页 |
| 1.10 Proteomics | 第44-46页 |
| 1.11 Plant Proteomics | 第46-47页 |
| 1.12 Application of proteomics in studying plant stress responses mechanisms | 第47页 |
| 1.13 Objectives of this study | 第47-48页 |
| Chapter 2. Overexpression of TERF1 and TSRF1 improve drought and salt tolerance in sugarcane | 第48-77页 |
| 2.1 Summary | 第48页 |
| 2.2 Introduction | 第48-50页 |
| 2.3 Materials and methods | 第50-55页 |
| 2.3.1 Construction of plasmid vectors containing TERF1 and TSRF1 gene and bacterial | 第50页 |
| 2.3.2 Embryogenic callus induction and culture | 第50-51页 |
| 2.3.3 Preparation of Agrobacterium suspension | 第51页 |
| 2.3.4 Callus infection, co-cultivation and regeneration of transformed plantlets | 第51页 |
| 2.3.5 Hardening of the plantlets | 第51-52页 |
| 2.3.6 Molecular analysis of putative transgenic plants | 第52-53页 |
| 2.3.7 Drought and salt stress treatment | 第53页 |
| 2.3.8 Determination of chlorophyll content | 第53页 |
| 2.3.9 Estimation of relative water content (RWC) | 第53页 |
| 2.3.10 Determination of proline content | 第53-54页 |
| 2.3.11 Estimation of total soluble sugars (TSS) | 第54页 |
| 2.3.12 Estimation of lipid peroxidation | 第54-55页 |
| 2.3.13 Determination of glycine betaine | 第55页 |
| 2.3.14 Hydrogen peroxide (H_2O_2) estimation | 第55页 |
| 2.3.15 Statistics | 第55页 |
| 2.4 Results | 第55-72页 |
| 2.4.1 Induction of embryogenic callus | 第55-56页 |
| 2.4.2 Effect of inoculum density on transformation | 第56页 |
| 2.4.3 Effect of co-cultivation media and co-cultivation period on transformation | 第56-57页 |
| 2.4.4 Plant regeneration | 第57-58页 |
| 2.4.5 Screening of putative transgenic plantlets by PCR | 第58-60页 |
| 2.4.6 Response of transgenic lines under drought stress | 第60-61页 |
| 2.4.7 Response of transgenic lines under salt stress | 第61页 |
| 2.4.8 Drought induced changes of relative water and chlorophyll content in transgenic | 第61-64页 |
| 2.4.9 Overexpression of TERF1 increases proline, soluble sugars and glycine betaine content insugarcane under drought stress | 第64-67页 |
| 2.4.10 Overexpression of TERF1 accumulates less MDA and H_2O_2 in sugarcane under droughtstress | 第67页 |
| 2.4.11 Overexpression of TERF1 increases proline,soluble sugars and glycine betaine content insugarcane under salt stress | 第67-68页 |
| 2.4.12 Overexpression of TERF1 accumulates less MDA and H_2O2_ in Sugarcane under salt | 第68-70页 |
| 2.4.13 Overexpression of TSRF1 increases proline,soluble Sugars and glycine betaine content insugarcane under drought stress | 第70-71页 |
| 2.4.14 Overexpression of TSRF1 accumulates less MDA and H_2O_2 in sugarcane under droughtstress | 第71-72页 |
| 2.4.15 Overexpression of TSRF1 increases proline,soluble sugars and glycine betaine content insugarcane under salt stress | 第72页 |
| 2.4.16 Overexpression of TSRF1 accumulates less MDA and H_2O_2 in sugarcane under salt | 第72页 |
| 2.5 Discussions | 第72-76页 |
| 2.6 Conclusions | 第76-77页 |
| Chapter 3. Proteomic analysis of drought stress responsive protein in TERF1 transgenic | 第77-105页 |
| 3.1 Summary | 第77-78页 |
| 3.2 Introduction | 第78-79页 |
| 3.3 Materials and Methods | 第79-82页 |
| 3.3.1 Drought stress treatment | 第79页 |
| 3.3.2 Extraction of protein from sugarcane leaves | 第79页 |
| 3.3.3 Protein lysis and quantification | 第79-80页 |
| 3.3.4 First-dimension isoelectric focusing(IEF) | 第80页 |
| 3.3.5 Two-dimensional electrophoresis(2-DE) | 第80页 |
| 3.3.6 Protein visualization | 第80-81页 |
| 3.3.7 Image analyses | 第81页 |
| 3.3.8 Spot picking MALDI-TOF-MS and database search | 第81页 |
| 3.3.9 Bioinformatics analysis of differentially expressed proteins | 第81-82页 |
| 3.4 Results | 第82-96页 |
| 3.4.1 Effect of drought stress on protein content of sugarcane leaves | 第82页 |
| 3.4.2 Identification of differentially expressed protein under drought stress | 第82-92页 |
| 3.4.3 Functional classification of differentially expressed proteins | 第92-94页 |
| 3.4.4 Physical and chemical properties of differentially expressed proteins | 第94-96页 |
| 3.5 Discussion | 第96-103页 |
| 3.5.1 Expression changes of metabolism related proteins | 第96-98页 |
| 3.5.2 Expression changes of energy related proteins | 第98-100页 |
| 3.5.3 Expression changes of protein synthesis related proteins | 第100-101页 |
| 3.5.4 Expression changes of Transporter proteins | 第101页 |
| 3.5.5 Expression changes of Transcription proteins | 第101页 |
| 3.5.6 Expression changes of intracellular traffic proteins | 第101-102页 |
| 3.5.7 Expression changes of signal transduction proteins | 第102页 |
| 3.5.8 Expression changes of defense proteins | 第102-103页 |
| 3.6 Conclusions | 第103-105页 |
| Chapter 4. General conclusion | 第105-106页 |
| References | 第106-131页 |
| Publication at PhD study | 第131-132页 |
| Acknowledgements | 第132-133页 |
| Resume | 第133页 |
