| 摘要 | 第6-8页 |
| Abstract | 第8-9页 |
| Abbreviation | 第19-20页 |
| Chapter 1:Introduction | 第20-37页 |
| 1.1 Malnutrition in the world | 第20-22页 |
| 1.1.1 Economic impact of malnutrition | 第20页 |
| 1.1.2 Causes of malnutrition/micronutrients deficiencies | 第20-21页 |
| 1.1.3 Strategies to combat the malnutrition | 第21-22页 |
| 1.2 Wheat as a potential crop for biofortification | 第22-23页 |
| 1.3 Folates biosynthesis and biofortification | 第23-30页 |
| 1.3.1 Folate biosynthesis | 第23-27页 |
| 1.3.2 Problems caused by folate deficiency | 第27页 |
| 1.3.3 Strategies to fight against folate deficiency | 第27-30页 |
| 1.4 Anthocyanin biosynthesis and biofortification | 第30-35页 |
| 1.4.1 Anthocyanins and their nutraceuticals properties | 第30页 |
| 1.4.2 Anthocyanin biosynthesis | 第30-34页 |
| 1.4.3 Biofortification for anthocyanins | 第34-35页 |
| 1.5 Objectives of the present study | 第35-37页 |
| Chapter 2:Folate content analysis of wheat cultivars developed in the North China Plain | 第37-51页 |
| 2.1 Introduction | 第37-38页 |
| 2.2 Materials and methods | 第38-41页 |
| 2.2.1 Plant materials | 第38-39页 |
| 2.2.2 Chemicals and reagents | 第39页 |
| 2.2.3 Folate extraction and deglutamylation | 第39-40页 |
| 2.2.4 Folate determination by HPLC–MS/MS | 第40页 |
| 2.2.5 Statistical analysis | 第40-41页 |
| 2.3 Results | 第41-47页 |
| 2.3.1 Overall variation of the total folate levels in wheat samples | 第41-42页 |
| 2.3.2 Screening of wheat genotypes with high folate content in different regions | 第42-43页 |
| 2.3.3 Distribution of folate forms in Chinese wheat genotypes | 第43-46页 |
| 2.3.4 Association of wheat folate contents with environment | 第46-47页 |
| 2.4 Discussion | 第47-50页 |
| 2.5 Summary | 第50-51页 |
| Chapter 3:Folate fortification of wheat by genetic engineering approach | 第51-70页 |
| 3.1 Introduction | 第51-53页 |
| 3.2 Materials and methods | 第53-58页 |
| 3.2.1 Plant material and growth conditions | 第53页 |
| 3.2.2 Construction and transformation of overexpression vectors | 第53-54页 |
| 3.2.3 Identification of transgenic wheat plants through Quickstix method | 第54页 |
| 3.2.4 Identification of transgenic plants through PCR amplification of transgenes | 第54-55页 |
| 3.2.5 Determination of expression levels of transgenes in the grains of transgenic plants | 第55页 |
| 3.2.6 Obtaining of homozygous wheat transgenic plants through double haploids | 第55-56页 |
| 3.2.7 Chromosome preparation and fluorescent in situ hybridization | 第56-57页 |
| 3.2.8 Determination of levels of folate and its precursors contained in transgenic plants | 第57-58页 |
| 3.2.9 Investigation the effect of transgenes GmGCHI and GmADCS on agronomic traits of transgenic wheat plants | 第58页 |
| 3.2.10 Statistical analysis | 第58页 |
| 3.3 Results | 第58-68页 |
| 3.3.1 Co-expression of GmGCHI and GmADCS in wheat transgenic plants | 第58-63页 |
| 3.3.2 Co-expression of codon-optimized soybean GmGCHI and tomato LeADCS in wheat | 第63-68页 |
| 3.4 Discussion | 第68-69页 |
| 3.5 Summary | 第69-70页 |
| Chapter 4:Anthocyanin accumulation in wheat through expression of maize transcriptional factors | 第70-94页 |
| 4.1 Introduction | 第70-71页 |
| 4.2 Materials and methods | 第71-77页 |
| 4.2.1 Plant material and growth conditions | 第71-72页 |
| 4.2.2 Construction of expression vectors containing transcriptional factors involved in anthocyanin biosynthesis | 第72-73页 |
| 4.2.3 Agrobacterium-mediated transformation using wheat immature embryos | 第73-74页 |
| 4.2.4 Detection of bar protein through Quickstix strip method | 第74页 |
| 4.2.5 DNA extraction and PCR amplification | 第74页 |
| 4.2.6 Southern blot analysis | 第74-75页 |
| 4.2.7 Chromosome preparation and fluorescent in situ hybridization | 第75页 |
| 4.2.8 Obtaining of homozygous wheat transgenic plants through double haploids | 第75-76页 |
| 4.2.9 RNA extraction and quantitative real-time PCR assay | 第76页 |
| 4.2.10 Determination of pigment contents in the transgenic wheat plants | 第76-77页 |
| 4.3 Results | 第77-90页 |
| 4.3.1 Purple phenotype of expressed R2R3-MYB and bHLH type TFs in wheat immature embryos and derived tissues after transformation | 第77-78页 |
| 4.3.2 Agrobacterium-mediated transformation efficiency and obtaining of wheat transgenic plants | 第78-80页 |
| 4.3.3 Quickstix detection of bar protein in wheat transgenic wheat plants | 第80页 |
| 4.3.4 PCR detection of transgenic wheat plants | 第80页 |
| 4.3.5 Southern blot analysis | 第80-81页 |
| 4.3.6 Production of doubling haploid wheat plants through gynogenesis | 第81-82页 |
| 4.3.7 Production of doubling haploid plants through colchicine application | 第82页 |
| 4.3.8 Identification of stable transgenic wheat plants through fluorescent in situ hybridization analysis(FISH) | 第82-83页 |
| 4.3.9 Phenotype of three types of stable transgenic lines expressing ZmC1 and/or ZmR genes | 第83-87页 |
| 4.3.10 Expression profiling of the two target genes ZmC1 and ZmR as well as their wheat homologous genes in the three types of transgenic lines | 第87-88页 |
| 4.3.11 Expression profiling of wheat native anthocyanin biosynthesis related genes in the three types of transgenic lines | 第88-90页 |
| 4.3.12 Pigment contents in the seeds of transgenic wheat plants | 第90页 |
| 4.4 Discussion | 第90-92页 |
| 4.5 Summary | 第92-94页 |
| Conclusion | 第94-96页 |
| References | 第96-109页 |
| ACKNOWLEDGEMENTS | 第109-110页 |
| RESUME | 第110-112页 |
| 附件 | 第112-131页 |
