| 摘要 | 第6-7页 |
| Abstract | 第7-8页 |
| Chapter 1 REVIEW | 第18-47页 |
| 1.0 General introduction | 第18-20页 |
| 1.1 Soybean origin,classification, domestication and distribution | 第20-23页 |
| 1.2 Soybeanplant breeding | 第23-28页 |
| 1.2.1 Conventional soybean breeding | 第23-25页 |
| 1.2.2 Soybean molecular breeding | 第25-28页 |
| 1.3 Advances of soybean plant genetics | 第28-31页 |
| 1.4 Novel allele discovery | 第31-45页 |
| 1.4.1 TILLING | 第32-42页 |
| 1.4.1.1 Employment of TILLING in crops and model plants | 第33-41页 |
| 1.4.1.2 Employment of TILLING in soybean | 第41-42页 |
| 1.4.2 EcoTILLING | 第42-45页 |
| 1.4.2.1 Employment of EcoTILLING in crops and model plants | 第42-45页 |
| 1.5 Objectives of the study | 第45-47页 |
| Chapter 2 DETECTION OF INDUCED MUTATIONS RELATED TO SOYBEAN SEED SIZE ANDMATURITY BY TILLING | 第47-89页 |
| 2.1 Introduction | 第48-51页 |
| 2.2 Material and Methods | 第51-58页 |
| 2.2.1 Development of mutagenesis population | 第51-52页 |
| 2.2.2 TILLING procedure | 第52-58页 |
| 2.2.2.1 DNA preparations | 第53-54页 |
| 2.2.2.2 Designing PCR primers for target genes for TILLING and EcoTILLING | 第54-57页 |
| 2.2.2.3 Preparation of CEL1 for detection heteroduplexes | 第57-58页 |
| 2.2.2.4 Fragment separation of digested PCR products | 第58页 |
| 2.3 Results | 第58-82页 |
| 2.3.1 Detection of IKU2 alleles | 第58-67页 |
| 2.3.1.1 Identification and description of IKU2 gene of soybean | 第58-59页 |
| 2.3.1.2 Optimization of CEL1 digestion condition and estimation of mutation rate | 第59-61页 |
| 2.3.1.3 Allele variation of IKU2 gene in the mutagenesis population | 第61-65页 |
| 2.3.1.4 The detected haplotypes based nucleotides and amino acids in IKU2 gene | 第65-67页 |
| 2.3.2 TILLING of E4 and E3 gene in soybean mutagenesis population | 第67-81页 |
| 2.3.2.1 Identification of E3 and E4 gene | 第67页 |
| 2.3.2.2 CEL1 digestion and fragment separation of E4 and E3 primers | 第67-70页 |
| 2.3.2.3 Allele variation of E3 and E4 gene in the mutagenesis population | 第70-79页 |
| 2.3.2.4 The detected haplotypes based nucleotides and amino acids in E4 gene | 第79-81页 |
| 2.3.3 Summary of detected mutations of IKU2, E3 and E4 in mutagenesis population | 第81-82页 |
| 2.4 Discussions | 第82-87页 |
| 2.4.1 Development of mutant population | 第82-83页 |
| 2.4.2 TILLING of IKU2 gene in mutagenesis soybean | 第83-85页 |
| 2.4.2.1 Identification and description of IKU2 gene | 第83页 |
| 2.4.2.2 Detected mutations in IKU2 in mutagenesis soybean | 第83-84页 |
| 2.4.2.3 Identification of induced mutation of IKU2 in mutagenesis soybean | 第84-85页 |
| 2.4.3 TILLING of E4 and E3 gene in mutagenesis soybean | 第85-87页 |
| 2.4.3.1 CEL1 Digestion of E4 and E3 primers in mutagenesis soybean | 第85-87页 |
| 2.5 Conclusion | 第87-89页 |
| Chapter 3 ECOTILLING TO DISCOVER NATURAL VARIATIONS OF IKU2,E3 AND E4 CANDIDATE GENES RELATED TO SEED SIZE AND MATURITY IN NATURALSOYBEAN | 第89-125页 |
| 3.1 Introduction | 第90-95页 |
| 3.2 Material and methods | 第95-96页 |
| 3.2.1 Germplasm | 第95页 |
| 3.2.2 EcoTILLING Procedure | 第95-96页 |
| 3.2.2.1 DNA preparations | 第95页 |
| 3.2.2.2 Designing of primers and PCR amplification | 第95页 |
| 3.2.2.3 Preparation of CJE for detection heteroduplexes | 第95-96页 |
| 3.2.2.4. Fragment separation of digested PCR products | 第96页 |
| 3.3 Results | 第96-117页 |
| 3.3.1 EcoTILLING in IKU2 gene of soybean | 第96-105页 |
| 3.3.1.1 CEL1 digestion of IKU2 primers and fragment separation | 第96-102页 |
| 3.3.1.2 Identification of DNA nucleotide variations of IKU2 in soybean naturalpopulation | 第102-103页 |
| 3.3.1.3 UPGMA Phylogenic tree of 27 soybean accessions | 第103-105页 |
| 3.3.2 EcoTILLING in E4 and E3 gene of soybean | 第105-116页 |
| 3.3.2.1 CEL1 digestion and fragment separation of E4 and E3 primers utilized innatural population | 第105-113页 |
| 3.3.2.2 The detected haplotypes based nucleotides and amino acids in E4 gene | 第113-115页 |
| 3.3.2.3 UPGMA Phylogenic tree of 15 soybean accessions | 第115-116页 |
| 3.3.3. Summary of the detected mutations of IKU2, E3 and E4 in soybean naturalpopulation through EcoTILLING | 第116-117页 |
| 3.4 Discussions | 第117-124页 |
| 3.4.1 Germplasm | 第117-118页 |
| 3.4.2 EcoTILLING of IKU2 gene in soybean natural population | 第118-121页 |
| 3.4.2.1 Identification and description of IKU2 gene | 第118页 |
| 3.4.2.2 CEL1 Digestion of IKU2 primers in soybean natural population | 第118-119页 |
| 3.4.2.3 Identification of nucleotide variations of IKU2 gene in soybean naturalpopulation | 第119-120页 |
| 3.4.2.4 UPGMA phylogeny tree of detected variations in soybean naturalpopulation | 第120-121页 |
| 3.4.3 EcoTILLING of E4 and E3 gene in soybean natural population | 第121-124页 |
| 3.4.3.1 Identification of E4 and E3 gene | 第121-124页 |
| 3.5 Conclusion | 第124-125页 |
| Chapter 4 COMPARISONS BETWEEN TILLING AND ECOTILLING SCREENS OF IKU2,E3 AND E4 GENES IN MUTAGENESIS AND NATURAL SOYBEANS | 第125-136页 |
| 4.1 Introduction | 第126-127页 |
| 4.3 Results and discussions | 第127-132页 |
| 4.3.1 Detected mutations in mutagenesis and natural populations | 第127-128页 |
| 4.3.2 Nucleotide variations in mutagenesis and natural populations | 第128-130页 |
| 4.3.3 The 'common' and 'private' nucleotide variations | 第130-131页 |
| 4.3.4 The phenotypes of detected mutagenesis and natural soybeans | 第131-132页 |
| 4.4 Discussions | 第132-135页 |
| 4.4.1 Detected mutations in mutagenesis and natural soybean | 第132-133页 |
| 4.4.2 'Common' and ‘private’ nucleotide variations in mutagenesis and untreated soybean | 第133-134页 |
| 4.4.3 The phenotypes of detected mutagenesis and natural soybeans | 第134-135页 |
| 4.5 Conclusions | 第135-136页 |
| Chapter 5 OVERALL CONCLUSION | 第136-138页 |
| 5.1 Detection of mutation by Tilling | 第136页 |
| 5.2 Detection of natural variation by Ecotilling | 第136页 |
| 5.3 Comparison of TILLING and EcoTILLING | 第136-137页 |
| 5.4 Prospect | 第137-138页 |
| Reference | 第138-152页 |
| ACKNOWLEDGEMENT | 第152-153页 |
| RESUME | 第153-157页 |
