大豆中油份合成调控相关转录因子的功能研究
发布时间：2025-07-08 23:48
大豆(Glycinemax)是一种豆科作物,在豆类生产中较大范围的商业应用因此得以在全世界范围内广泛种植。大豆油是人类食物的重要组成部分,并且是众多工业生产应用的原材料。脂质合成和代谢是生命系统的主要活动,例如,三酰甘油(TAG)是储存物质,而磷脂在生物膜的形成中发挥着至关重要的作用,而生物膜在植物早期发育和生长中起着关键作用,同时磷脂作为食物成分也为人类提供了营养来源。传统的(育种)方法是通过调控油脂合成途径中的关键基因来实现的。然而这种方法的局限性在于只能够调控脂肪酸合成、降解过程特定的某一个基因。克服这一缺点的方法,就是通过操纵潜在的转录因子,激活调控油脂合成复合途径中的多个基因。本研究的主要目的,即通过操纵ABSCISIC ACID-INSENSITIVE3(ABI3),FUSCA3(FUS3)和LEAFYCOTYLEDON2(LEC2;AFL)等B3结构域的转录因子和HAP3结构域转录因子LEC1提高植物组织中的油份;这些转录因子在种子发育以及种子中代谢物诸如蛋白质、碳水化合物和脂质储存过程中发挥重要作用。我们克隆了大豆中的这些转录因子,通过GATEWAY体系构建到p B2GW...
【文章页数】：113 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER Ⅰ. LITERATURE REVIEW
1.1 General background
1.2 Lipid biosynthesis
1.2.1 FA biosynthesis
1.2.2 Galactolipid biosynthesis in the chloroplast
1.2.3 Neutral and polar lipids biosynthesis in the endoplasmic reticulum
1.3 TAG storage in oil body
1.4 The transporters involved in lipid degradation and beta oxidation
1.5 Cross-membrane lipid transport and trafficking
1.5.1 Flip-flop movements
1.5.2 Plastid-ER FA transport
1.5.3 ER-Plastid phospholipid and FA transport
1.6 Transporters required for surface lipid deposition
1.7 Role of transcriptional regulation in lipid accumulation
1.7.1 Leafy cotyledon 1 (LEC1)
1.7.2 Leafy cotyledon 2 (LEC2)
1.7.3 Abscisic acid insensitive (ABI3) and FUSCA3 (FUS3)
1.7.4 Wrinkled 1 (WRI1)
1.7.5 Dofs increase the total FA content in transgenic plants
1.7.6 Role of Basic leucine zipper (b ZIPs) in lipid accumulation
1.7.7 MYB TFs regulate biosynthesis of VLCFAs
1.7.8 Negative regulators of oil biosynthesis
1.8 Biotechnology tools for metabolic engineering of oil yield and composition
1.9 Economical importance of soybean crop
1.10 Aims and objectives of the current study
CHAPTER Ⅱ. MATERIALS AND METHODS
2.1 Identification of the soybean LAFL gene and in silico analysis
2.2 Phylogenetic analysis
2.3 Gene cloning
2.4 Gel electrophoresis and purification of PCR product
2.5 Arabidopsis thaliana transformation
2.6 Plant growth conditions and treatment
2.7 RNA extraction
2.8 Quantitative RT-PCR (q RT-PCR) analysis of gene expression
2.9 Genomic DNA extraction
2.10 Plasmid extraction
2.11 Seed oil extraction
2.12 Hairy root induction
2.13 Hairy root oil extraction
2.14 Histochemical staining for starch
2.15 Spectrophotometric determination of starch
2.16 Protein extraction
2.17 Anthocynin measurement
2.18 CDNA library construction for Illumina deep sequencing
2.19 Statistic analysis
CHAPTER Ⅲ. RESULTS
3.1 Identification of LAFL genes in Glycine max genome
3.2 Expression of Gm LAFL during soybean seed development
3.3 Expression in Arabidopsis thaliana mutants alters seed oil production and TAG composition
3.4 Ectopic expression in Arabidopsis thaliana modifies seed oil production and TAG composition
3.5 GmLAFL genes alters the chemical composition of seeds
3.6 GmLEAFY ectopic expression increases total protein in Arabidopsis seeds
3.7 GmLEAFY genes are responsible for embryonic-vegetative phase transition
3.8 Ectopic expression in soybean hairy roots promotes TAG accumulation
3.9 Transcriptome analysis of over expressed soybean hairy roots
CHAPTER Ⅳ. DISCUSSION
4.1 Each member of LAFL-clade regulate TAG content and composition
4.2 GmLEC2 regulates starch and protein metabolism
4.3 GmLEC2 and GmFUS3 target key regulatory genes in oil biosynthesis
4.4 GmLEC2 and GmFUS3 target key genes involved in FA and TAG biosynthesis
4.5 Genes involved in TAG catabolism and lipid hydrolysis are regulated by GmLEC2 and GmFUS3
4.6 GmFUS3 and GmLEC2 control the regulation of lipid transporters
CHAPTER Ⅴ. CONCLUSIONS
REFERENCES
APPENDIX A. LIST OF PRIMERS USED IN THIS STUDY
APPENDIX B. PREDICTED EXPRESSION OF LAFL GENES IN DIFFERENT TISSUES OF SOYBEAN IN PUBLIC DATABASE (PHYTOZOME)
APPENDIX C. PHYLOGENETIC ANALYSIS OF GMFUS3 AND GMLEC2 TARGETS IN G.MAX AND THEIR ORTHOLOGS IN OTHER CROPS
APPENDIX D. LIST OF TRANSCRIPTION FACTORS AND TRANPORTERS REGULATED BY GMLEC2 AND GMFUS3 IN LIPID BIOSYNTHESIS PATHWAY
ACKNOWLEDGEMENT
PUBLICATIONS
本文编号：4056807
【文章页数】：113 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
LIST OF ABBREVIATIONS
CHAPTER Ⅰ. LITERATURE REVIEW
1.1 General background
1.2 Lipid biosynthesis
1.2.1 FA biosynthesis
1.2.2 Galactolipid biosynthesis in the chloroplast
1.2.3 Neutral and polar lipids biosynthesis in the endoplasmic reticulum
1.3 TAG storage in oil body
1.4 The transporters involved in lipid degradation and beta oxidation
1.5 Cross-membrane lipid transport and trafficking
1.5.1 Flip-flop movements
1.5.2 Plastid-ER FA transport
1.5.3 ER-Plastid phospholipid and FA transport
1.6 Transporters required for surface lipid deposition
1.7 Role of transcriptional regulation in lipid accumulation
1.7.1 Leafy cotyledon 1 (LEC1)
1.7.2 Leafy cotyledon 2 (LEC2)
1.7.3 Abscisic acid insensitive (ABI3) and FUSCA3 (FUS3)
1.7.4 Wrinkled 1 (WRI1)
1.7.5 Dofs increase the total FA content in transgenic plants
1.7.6 Role of Basic leucine zipper (b ZIPs) in lipid accumulation
1.7.7 MYB TFs regulate biosynthesis of VLCFAs
1.7.8 Negative regulators of oil biosynthesis
1.8 Biotechnology tools for metabolic engineering of oil yield and composition
1.9 Economical importance of soybean crop
1.10 Aims and objectives of the current study
CHAPTER Ⅱ. MATERIALS AND METHODS
2.1 Identification of the soybean LAFL gene and in silico analysis
2.2 Phylogenetic analysis
2.3 Gene cloning
2.4 Gel electrophoresis and purification of PCR product
2.5 Arabidopsis thaliana transformation
2.6 Plant growth conditions and treatment
2.7 RNA extraction
2.8 Quantitative RT-PCR (q RT-PCR) analysis of gene expression
2.9 Genomic DNA extraction
2.10 Plasmid extraction
2.11 Seed oil extraction
2.12 Hairy root induction
2.13 Hairy root oil extraction
2.14 Histochemical staining for starch
2.15 Spectrophotometric determination of starch
2.16 Protein extraction
2.17 Anthocynin measurement
2.18 CDNA library construction for Illumina deep sequencing
2.19 Statistic analysis
CHAPTER Ⅲ. RESULTS
3.1 Identification of LAFL genes in Glycine max genome
3.2 Expression of Gm LAFL during soybean seed development
3.3 Expression in Arabidopsis thaliana mutants alters seed oil production and TAG composition
3.4 Ectopic expression in Arabidopsis thaliana modifies seed oil production and TAG composition
3.5 GmLAFL genes alters the chemical composition of seeds
3.6 GmLEAFY ectopic expression increases total protein in Arabidopsis seeds
3.7 GmLEAFY genes are responsible for embryonic-vegetative phase transition
3.8 Ectopic expression in soybean hairy roots promotes TAG accumulation
3.9 Transcriptome analysis of over expressed soybean hairy roots
CHAPTER Ⅳ. DISCUSSION
4.1 Each member of LAFL-clade regulate TAG content and composition
4.2 GmLEC2 regulates starch and protein metabolism
4.3 GmLEC2 and GmFUS3 target key regulatory genes in oil biosynthesis
4.4 GmLEC2 and GmFUS3 target key genes involved in FA and TAG biosynthesis
4.5 Genes involved in TAG catabolism and lipid hydrolysis are regulated by GmLEC2 and GmFUS3
4.6 GmFUS3 and GmLEC2 control the regulation of lipid transporters
CHAPTER Ⅴ. CONCLUSIONS
REFERENCES
APPENDIX A. LIST OF PRIMERS USED IN THIS STUDY
APPENDIX B. PREDICTED EXPRESSION OF LAFL GENES IN DIFFERENT TISSUES OF SOYBEAN IN PUBLIC DATABASE (PHYTOZOME)
APPENDIX C. PHYLOGENETIC ANALYSIS OF GMFUS3 AND GMLEC2 TARGETS IN G.MAX AND THEIR ORTHOLOGS IN OTHER CROPS
APPENDIX D. LIST OF TRANSCRIPTION FACTORS AND TRANPORTERS REGULATED BY GMLEC2 AND GMFUS3 IN LIPID BIOSYNTHESIS PATHWAY
ACKNOWLEDGEMENT
PUBLICATIONS
本文编号：4056807
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