| 摘要 | 第6-7页 |
| abstract | 第7-8页 |
| LIST OF ABBREVIATION | 第15-17页 |
| Chapter 1 Introduction | 第17-26页 |
| 1.1 Wheat domestication and evolution | 第17页 |
| 1.2 Wheat importance and production | 第17页 |
| 1.3 Impact of abiotic stresses in crops | 第17-18页 |
| 1.4 Wheat plants strategies to tackle drought stress | 第18-19页 |
| 1.5 ABA hormone and its role under abiotic stress | 第19-20页 |
| 1.6 ABA receptors in plants | 第20-21页 |
| 1.7 Structure and function of PYL | 第21-22页 |
| 1.8 Role of PYL in ABA signaling pathway | 第22页 |
| 1.9 Role of PYL in gene expression and transcriptional responses under abiotic stress | 第22-24页 |
| 1.10 Significance of research work | 第24页 |
| 1.11 Research objective | 第24-26页 |
| Chapter 2 TaPYL3 expression and bioinformatics analyses | 第26-41页 |
| 2.1 Introduction to Quantitative Real-Time PCR | 第26-27页 |
| 2.1.1 Gene regulation pattern in PYL receptors | 第26页 |
| 2.1.2 Bioinformatics analysis in gene families | 第26-27页 |
| 2.1.3 Research objectives | 第27页 |
| 2.2 Materials and methods | 第27-30页 |
| 2.2.1 Plant materials | 第27页 |
| 2.2.2 Treatment and conditions | 第27页 |
| 2.2.3 RNA extraction | 第27-28页 |
| 2.2.4 Quantification of RNA on nano-drop spectrophotometer | 第28页 |
| 2.2.5 c DNA synthesis | 第28-29页 |
| 2.2.6 Quantitative real-time PCR (q RT- PCR) | 第29页 |
| 2.2.7 Primers designed for gene expression analyses by q RT-PCR | 第29-30页 |
| 2.2.8 Sequence analysis and alignment of Ta PYL3 | 第30页 |
| 2.2.9 Phylogenetic tree construction | 第30页 |
| 2.3 Results | 第30-38页 |
| 2.3.1 Ta PYL3 sequence alignment | 第30-33页 |
| 2.3.2 Phylogenetic analysis | 第33-34页 |
| 2.3.3 Ta PYL3 response to PEG treatment | 第34页 |
| 2.3.4 Ta PYL3 response to Na Cl treatment | 第34-35页 |
| 2.3.5 Ta PYL3 response to cold treatment | 第35-36页 |
| 2.3.6 Ta PYL3 response to heat stress | 第36-38页 |
| 2.3.7 Ta PYL3 response to ABA treatment | 第38页 |
| 2.4 Discussion | 第38-40页 |
| 2.5 Conclusion | 第40-41页 |
| Chapter 3 Abiotic stress tolerance in transgenic Arabidopsis | 第41-59页 |
| 3.1 Introduction | 第41-42页 |
| 3.1.1 Phenotypes of transgenic plants under abiotic stress | 第41-42页 |
| 3.2 Materials and Methods | 第42-53页 |
| 3.2.1 Transgenic plant material cultivation | 第42页 |
| 3.2.2 Isolation of Ta PYL3 candidate sequence | 第42页 |
| 3.2.3 Primers used | 第42页 |
| 3.2.4 Extraction of wheat genomic DNA | 第42-43页 |
| 3.2.5 PCR and agarose gel electrophoresis | 第43页 |
| 3.2.6 Purification of PCR product | 第43-44页 |
| 3.2.7 Cloning and heat shock transformation | 第44页 |
| 3.2.8 Heat shock transformation protocol | 第44-45页 |
| 3.2.9 Colony PCR to confirm the positive plasmid with gene insert | 第45页 |
| 3.2.10 Plasmid extraction | 第45-46页 |
| 3.2.11 Plasmid DNA sequencing | 第46-48页 |
| 3.2.12 PCR for generating plasmid vector construct | 第48-50页 |
| 3.2.13 Preparation of competent cells of A. tumefaciens | 第50页 |
| 3.2.14 Agrobacterium mediated transformation in Arabidopsis by floral dip method | 第50页 |
| 3.2.15 Screening for selection of transformed Arabidopsis seeds | 第50-51页 |
| 3.2.16 Seed sterilization | 第51页 |
| 3.2.17 Preparation of MS nutrient medium | 第51页 |
| 3.2.18 Tissue culture | 第51页 |
| 3.2.19 Abiotic stress treatment of Ta PYL3 transformed Arabidopsis plants | 第51-52页 |
| 3.2.20 Gene amplification and transcriptional expression in transgenic lines | 第52-53页 |
| 3.2.21 Relative expression level of transgenic Arabidopsis lines | 第53页 |
| 3.3 Results (Transgenic Arabidopsis) | 第53-57页 |
| 3.3.1 Mannitol stress treatment | 第53-54页 |
| 3.3.2 4°C stress treatment | 第54页 |
| 3.3.3 ABA stress treatment | 第54-56页 |
| 3.3.4 Na Cl stress treatment | 第56-57页 |
| 3.3.5 Gene amplification and relative expression in transgenic lines | 第57页 |
| 3.4 Discussion | 第57-58页 |
| 3.5 Conclusion | 第58-59页 |
| Chapter 4 Functional marker development of Ta PYL3 gene from wheat | 第59-78页 |
| 4.1 Introduction | 第59-60页 |
| 4.1.1 Development and application of molecular marker | 第59页 |
| 4.1.2 Natural variation in wheat by comparative genomics approach | 第59-60页 |
| 4.2 Materials and methods | 第60-62页 |
| 4.2.1 Plant materials | 第60页 |
| 4.2.2 Agronomic traits measurements of plant materials | 第60-61页 |
| 4.2.3 Primer specificity analysis | 第61页 |
| 4.2.4 Cloning and sequence analysis of Ta PYL3 | 第61页 |
| 4.2.5 Development of a functional marker for Ta PYL3 | 第61页 |
| 4.2.6 Gene linkage chromosome mapping | 第61-62页 |
| 4.2.7 Association analysis between Ta PYL3 allelic variation and agronomic traits | 第62页 |
| 4.3 Results | 第62-71页 |
| 4.3.1 Ta PYL3 genetic mapping | 第62-63页 |
| 4.3.2 Cloning and SNP identification of Ta PYL3 gene | 第63-64页 |
| 4.3.3 Functional marker development of Ta PYL3 | 第64-65页 |
| 4.3.4 Association of SNP (G/A) at 1605 position with chlorophyll content and spikelet number per spike | 第65-68页 |
| 4.3.5 Association of SNP ( C/A) at 2547 position with grains per spike, thousand kernel weight and yield per plant | 第68-70页 |
| 4.3.6 Geographical sharing of G/A SNP allelic variation in Chinese wheat producing regions | 第70页 |
| 4.3.7 Geographical sharing of C/A SNP allelic variation in Chinese wheat producing regions | 第70-71页 |
| 4.4 Discussions | 第71-78页 |
| 4.4.1 Role of Ta PYL3 in plant species | 第71页 |
| 4.4.2 TaPYL3 cloning and functional marker development | 第71-72页 |
| 4.4.3 TaPYL3 is a drought tolerance gene and may improve wheat yield and quality | 第72-73页 |
| 4.4.4 Selection of TaPYL3 alleles in wheat agro-ecological zones | 第73-78页 |
| Chapter 5 Conclusion | 第78-79页 |
| REFERENCES | 第79-92页 |
| ACKNOWLEDGEMENT | 第92-93页 |
| RESUME | 第93页 |
