Functional Characterization of Tomato Curl3 Gene in Metaboli
发布时间：2024-06-01 18:57
油菜素内酯(BRs)在植物生长发育中起着重要作用。然而,在园艺作物中,BR信号介导的植物生长调控的分子生物学证据仍然不足。BR信号转导与其他植物激素之间的互作共同调控植物生长发育。激素、环境信号和发育过程之间具有复杂互作机制,共同调控植物生长发育。本研究将有助于了解BR信号在植物生长发育调控中的作用。在本研究中,为了阐明BR信号传导对番茄生长发育的可能影响,我们基于CRISPR系统创建了BR不敏感型番茄curl3突变体。但是该突变体与对照的高度差异,我们同时从TGRC引进一个在curl3位点的等位基因突变体(abs1)。与curl3突变体相比,abs1突变体表型变化相对微弱。在本研究中,我们阐明了Sl Curl3(番茄中关键BR信号转导基因)在调节植物生长发育、初级代谢、激素平衡中的功能。主要结果如下:1)在本研究中,从番茄中分离了一个BRI1(油菜素内酯不敏感1基因)的同源基因Curl3,并基于CRISPR创建了敲除突变体,对其功能进行研究。在植物中BR信号突变体的功能丧失会导致其生理生化产生重要变化。curl3突变体表现出极度矮化,伴随着卷曲叶片以及生化和代谢途径的改变。此外,在对...
【文章页数】：110 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
List of Abbreviations
1 Introduction
1.1 Plant Hormones
1.2 Brassinolide Hormone
1.3 Brassinolide Crosstalk with other Hormones
1.4 Brassinolide Perception by BRI1
1.5 BR-Dependent Gene Expression
1.6 BR-Signaling Mutant
1.7 Objectives of This Research
2 Brassinolide Signaling Disruption Regulates Primary Metabolism in Accordance with Phytohormone Production
2.1 Materials and Methods
2.1.1 Plant Material
2.1.2 Gene Isolation
2.1.3 CRISPR/Cas9 Gene Editing
2.1.4 Morphological Characterization and Anatomical Observations
2.1.5 Hormone Quantification
2.1.6 Tomato Primary Metabolite Profiling
2.1.7 Quantitative Gene Expression Profiling
2.1.8 Statistical and Bioinformatics Analysis
2.2 Results
2.2.1 Sequence analysis of Curl3 from Solanum lycopersicum
2.2.2 CRISPR based Gene Editing
2.2.3 Altered Expression of BR Signaling Components
2.2.4 Morphological Characterization and Anatomical Observations of curl3 Mutant
2.2.5 BR-Insensitivity Alters Phytohormone Production in Transgenic Tomato Plants
2.2.6 Response of Primary Metabolites
2.2.7 Altered Expression of Metabolite Related Genes in curl3 Mutant
2.3 Discussion
2.4 Conclusion
3 Altered brassinolide sensitivity1 Transcriptionally Inhibits Chlorophyll Synthesis and Photosynthesis Capacity in Tomato
3.1 Materials and Methods
3.1.1 Plant Material and DNA Extraction
3.1.2 RNA-seq Library Preparation and Sequencing
3.1.3 Determination of Leaf Morphological Traits and Chlorophyll Accumulation
3.1.4 Chlorophyll Fluorescence Measurements
3.1.5 Photosynthetic Measurements
3.1.6 Gene Expression Quantification
3.1.7 Extraction and Quantification of Endogenous Phytohormones
3.1.8 Data Analysis
3.2 Results
3.2.1 abs1 Mutant Characterization
3.2.2 Altered Leaf Development in abs1 Mutant
3.2.3 BR-insensitive Mutant Modulates Endogenous Phytohormone Production
3.2.4 Reduced Chlorophyll Synthesis in abs1 Mutant
3.2.5 Chlorophyll Fluorescence Activity was Inhibited in abs1 Mutant
3.2.6 Photosynthetic Capacity was Inhibited in abs1 Mutant
3.2.7 Validation of DEGs
3.3 Discussion
3.4 Conclusion
4 Altered brassinolide sensitivity1 Impaired Floral Organ and Pollen Development
4.1 Materials and Methods
4.1.1 Plant Material and Growth Conditions
4.1.2 Quantitative Gene Expression Profiling
4.1.3 Extraction and Quantification of Phytohormones
4.1.4 Morphological Characterization and Anatomical Observations
4.1.5 Pollen Viability Assay
4.1.6 Seed Germination Assay
4.1.7 Statistical Analysis
4.2 Results
4.2.1 Temporal expression of Curl3 During Floral Development
4.2.2 abs1 Modulate BR and GA Related Genes and Endogenous Concentrations
4.2.3 abs1 Mutant Modulate Flower Development in Tomato
4.2.4 abs1 Mutant Modulate Flower Anatomical Structure in Tomato
4.2.5 BR Signaling Disruption Reduces Male Fertility
4.2.6 BR Signaling Disruption Reduces Seed Formation
4.3 Discussion
4.4 Conclusion
5 Altered brassinolide sensitivity1 Alters Fruit Size in Association with Phytohormones Modulation in Tomato
5.1 Materials and Methods
5.1.1 Plant material and Gene Isolation
5.1.2 Quantitative Gene Expression Profiling
5.1.3 Morphological Characterization and Anatomical Observations
5.1.4 Extraction and Quantification of Phytohormones
5.1.5 Statistical Analysis
5.2 Results
5.2.1 Temporal expression of Curl3 During Fruit Development
5.2.2 The Altered Transcript Level of BR Signaling Components in abs1 Mutant Fruit
5.2.3 BR Signaling Disruption Causes a Decrease in Phytohormone Production
5.2.4 BR Signaling Disruption Decreased Fruit Size of abs1 Mutant Fruit
5.2.5 BR Signaling Disruption Influences Cell Division and Expansion During Early Developmental Stages
5.2.6 The Altered Transcript Level of Fruit Size-Determining Genes in abs1 Mutant Fruit
5.3 Discussion
5.4 Conclusion
6 Summary and Future Perspectives
References
Appendices
Acknowledgements
本文编号：3986332
【文章页数】：110 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
List of Abbreviations
1 Introduction
1.1 Plant Hormones
1.2 Brassinolide Hormone
1.3 Brassinolide Crosstalk with other Hormones
1.4 Brassinolide Perception by BRI1
1.5 BR-Dependent Gene Expression
1.6 BR-Signaling Mutant
1.7 Objectives of This Research
2 Brassinolide Signaling Disruption Regulates Primary Metabolism in Accordance with Phytohormone Production
2.1 Materials and Methods
2.1.1 Plant Material
2.1.2 Gene Isolation
2.1.3 CRISPR/Cas9 Gene Editing
2.1.4 Morphological Characterization and Anatomical Observations
2.1.5 Hormone Quantification
2.1.6 Tomato Primary Metabolite Profiling
2.1.7 Quantitative Gene Expression Profiling
2.1.8 Statistical and Bioinformatics Analysis
2.2 Results
2.2.1 Sequence analysis of Curl3 from Solanum lycopersicum
2.2.2 CRISPR based Gene Editing
2.2.3 Altered Expression of BR Signaling Components
2.2.4 Morphological Characterization and Anatomical Observations of curl3 Mutant
2.2.5 BR-Insensitivity Alters Phytohormone Production in Transgenic Tomato Plants
2.2.6 Response of Primary Metabolites
2.2.7 Altered Expression of Metabolite Related Genes in curl3 Mutant
2.3 Discussion
2.4 Conclusion
3 Altered brassinolide sensitivity1 Transcriptionally Inhibits Chlorophyll Synthesis and Photosynthesis Capacity in Tomato
3.1 Materials and Methods
3.1.1 Plant Material and DNA Extraction
3.1.2 RNA-seq Library Preparation and Sequencing
3.1.3 Determination of Leaf Morphological Traits and Chlorophyll Accumulation
3.1.4 Chlorophyll Fluorescence Measurements
3.1.5 Photosynthetic Measurements
3.1.6 Gene Expression Quantification
3.1.7 Extraction and Quantification of Endogenous Phytohormones
3.1.8 Data Analysis
3.2 Results
3.2.1 abs1 Mutant Characterization
3.2.2 Altered Leaf Development in abs1 Mutant
3.2.3 BR-insensitive Mutant Modulates Endogenous Phytohormone Production
3.2.4 Reduced Chlorophyll Synthesis in abs1 Mutant
3.2.5 Chlorophyll Fluorescence Activity was Inhibited in abs1 Mutant
3.2.6 Photosynthetic Capacity was Inhibited in abs1 Mutant
3.2.7 Validation of DEGs
3.3 Discussion
3.4 Conclusion
4 Altered brassinolide sensitivity1 Impaired Floral Organ and Pollen Development
4.1 Materials and Methods
4.1.1 Plant Material and Growth Conditions
4.1.2 Quantitative Gene Expression Profiling
4.1.3 Extraction and Quantification of Phytohormones
4.1.4 Morphological Characterization and Anatomical Observations
4.1.5 Pollen Viability Assay
4.1.6 Seed Germination Assay
4.1.7 Statistical Analysis
4.2 Results
4.2.1 Temporal expression of Curl3 During Floral Development
4.2.2 abs1 Modulate BR and GA Related Genes and Endogenous Concentrations
4.2.3 abs1 Mutant Modulate Flower Development in Tomato
4.2.4 abs1 Mutant Modulate Flower Anatomical Structure in Tomato
4.2.5 BR Signaling Disruption Reduces Male Fertility
4.2.6 BR Signaling Disruption Reduces Seed Formation
4.3 Discussion
4.4 Conclusion
5 Altered brassinolide sensitivity1 Alters Fruit Size in Association with Phytohormones Modulation in Tomato
5.1 Materials and Methods
5.1.1 Plant material and Gene Isolation
5.1.2 Quantitative Gene Expression Profiling
5.1.3 Morphological Characterization and Anatomical Observations
5.1.4 Extraction and Quantification of Phytohormones
5.1.5 Statistical Analysis
5.2 Results
5.2.1 Temporal expression of Curl3 During Fruit Development
5.2.2 The Altered Transcript Level of BR Signaling Components in abs1 Mutant Fruit
5.2.3 BR Signaling Disruption Causes a Decrease in Phytohormone Production
5.2.4 BR Signaling Disruption Decreased Fruit Size of abs1 Mutant Fruit
5.2.5 BR Signaling Disruption Influences Cell Division and Expansion During Early Developmental Stages
5.2.6 The Altered Transcript Level of Fruit Size-Determining Genes in abs1 Mutant Fruit
5.3 Discussion
5.4 Conclusion
6 Summary and Future Perspectives
References
Appendices
Acknowledgements
本文编号：3986332
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