声明
Chapter 1 Introduction and Review of Literature
1.1 Background and Review of Literature
1.2 B3 Transcription Factor
1.2.1 ARF Family
1.2.2 LAV Family
1.2.3 RAV Family
1.2.4 REM Family
1.3 GASA Gene Family
1.4 FUS3 Role in Plant Growth and Development
1.4.1 LAFL Network
1.4.2 FUS3 Role in Embryo Development and Seed Maturation
1.4.3 FUS3 Role in Oil and Lipid Accumulation
1.4.4 FUS3 Role in Seed Germination and Dormancy
1.5 Research Objectives
Chapter 2 Genome-Wide Identification and Expression Profiling of B3 Transcription Factor Family in Grapevine during Different Stages of Ovule Development
2.1 Materials and Methods
2.1.1 Identification and Annotation of B3 Genes in Grapevine
2.1.2 Multiple Sequence Alignment and Phylogenetic Analysis
2.1.3 Analyses of Exon–Intron Structure, Distribution of Conserved Motifs and Characteristic Domain Architecture
2.1.4 Synteny Analysis of Grape, Tomato and Arabidopsis B3 Genes
2.1.5 Analysis of Cis-Acting Elements and Gene Ontology
2.1.6 Plant Materials
2.1.7 Main equipment/Instrument
2.1.8 Test Reagent
2.1.9 RNA Extraction and Synthesis of cDNA
2.1.10 Semi Quantitative Expression Analysis of Genes
2.1.11 Quantitative Real-time PCR Analysis of Selected Genes (RT-PCR)
2.2 Results
2.2.1 Genome-Wide Identification of B3 Genes in Grapevine
2.2.2 Phylogenetic Analysis of Grapes, Arabidopsis, and Tomato
2.2.3 Analysis of Gene Structure and Conserved Nucleotide Motifs
2.2.4 Domain Architecture Analysis of Grapevine B3 Proteins
2.2.5 Expansion Patterns of B3 Genes in Grape
2.2.6 Evolutionary Relationships among Grapevine, Tomato, and Arabidopsis B3 Genes
2.2.7 Expression Patterns of Grape B3 Genes during Seed Development
2.2.8 Developmental Regulation of Grapevine B3 Genes Outside of the Ovule
2.2.9 Cis-Acting Elements and Gene Ontology (Go) Analysis of B3 Genes
2.3 Discussion
2.3.1 Domain Description
2.3.2 Exon-Intron Analysis
2.3.3 Evolutionary Analysis
2.3.4 Synteny Analysis
2.3.5 Cis-elements and GO Analysis
2.3.6 Expression Profiling During Different Stages of Development
2.4 Summary
Chapter 3 Genomic Organization of GASA Gene Family in Grapevine and Expression Analysis During Different Stages of Seed Development in Seeded and Seedless Cultivars
3.1 Materials and Methods
3.1.1 Annotation and Identification of Putative Grapevine GASA Genes
3.1.2 Physicochemical Properties and Phylogeny Analysis
3.1.3 Exon–Intron, Gene Structures, Conserved Motif, and Promoter Analysis
3.1.4 Synteny Analysis and Calculation of Ka/Ks Ratio for Duplicated Genes
3.1.5 Plant Materials
3.1.6 Main equipment/Instrument
3.1.7 Test Reagent
3.1.8 Total RNA Extraction and cDNA synthesis
3.1.9 Primer Design and Expression Analysis by RT-PCR
3.2 Results
3.2.1 Genome-Wide Identification and Protein Features of GASA Genes in Grapevine
3.2.2 Phylogenetic Analysis of GASA Genes from Grape, Apple, and Arabidopsis
3.2.3 Analysis of Conserved Motifs, Domain Architecture, and Gene Structure
3.2.4 Grapevine Genes Duplication and Evolutionary Analysis
3.2.5 GASA Genes Expression Profiling During Seed Development
3.2.6 Tissue Specific Expression Profiling of Grapevine GASA Candidates
3.2.7 Promoter Analysis of GASA Genes
3.3 Discussion
3.3.1 Phylogenetic Analysis and Classification
3.3.2 Gene Duplication Analysis
3.3.3 Selection Pressure and Cis-element Analysis
3.3.4 Expression Analysis in Different Tissues at Different Developmental Stages
3.4 Summary
Chapter 4 Cloning and Functional Analysis of VvABI3-3
4.1 Materials and Methods
4.1.1 Plant Materials
4.1.2 Main Equipment/Instrument
4.1.3 Reagent Material
4.1.4 Primer for Gene Cloning
4.1.5 RNA Extraction and Reverse Transcription
4.1.6 Amplification and Sequence Analysis of Gene
4.1.7 DNA Recycling and Purification
4.1.8 Construction of Overexpression Vector
4.1.9 Genetic Transformation of Tomato Mediated by Agrobacterium GV3101
4.1.10 The Identification of Transgenic Tomatoes
4.1.11 Phenotypic Observations of Transgenic Tomato Plants
4.1.12 Histological Observations of Seed and Fruit
4.1.13 Pollen Viability and Microscopic Observation of Pollen Grains
4.1.14 Analysis of Gene Expression
4.1.15 Statistical Analysis
4.2 Results
4.2.1 Cloning of VvFUS3 and Identification of Transgenic Lines
4.2.2 Phenotypic Observations of Plant
4.2.3 Pollen Viability and Morphology Studies
4.2.4 Phenotypic and Histological Observations of Seed and Fruit
4.2.5 Expression Profiling of Genes Related with Fruit and Seed Development
4.2.6 Expression of GA and Auxin-related Genes is Differently Regulated in VvFUS3-OE Fruits
4.2.7 Expression Analysis of Genes directly Influenced by VvFUS3-OE Lines
4.2.8 Expression Analysis of Genes Affecting Fruit Size
4.3 Discussion
4.3.1 Amino Acid Sequence Alignment and Phylogenetic Analysis
4.3.2 Ectopic Expression VvFUS3 Influences Seed number and Size
4.3.3 Transgenic Tomatoes Influence Cell number and Size
4.3.4 VvFUS3 Influences Seed Development through Hormonal Signaling Pathways
4.4 Summary
Chapter 5 Conclusions and Future Prospective
5.1 Conclusions
5.2 Innovation and Future Prospective
参考文献
致谢
CURRICULUM VITAE
西北农林科技大学;
