Metabolic Engineering of Fatty Acid and Lipid Biosynthesis in Plant Seeds = 식물 종자 지질 생합성 경로의 대사공학 연구

목차 (Table of Contents)

• ABSTRACT
• TABLE OF CONTENTS
• LIST OF TABLES
• LIST OF FIGURES
• CHAPTER 1 Applications and prospects of genome editing in plant fatty
• acid and triacylglycerol biosynthesis
• 1.1. Abstract
• 1.2. Introduction
• 1.2.1. CRISPR/Cas9 and lipid metabolic engineering
• 1.2.2. Mutation in FAD2
• 1.2.3. Mutation in FATB and KASI
• 1.2.4. Mutation in FAE1
• 1.2.5. Mutation in acyltransferases
• 1.2.6. Mutation in phospholipases
• 1.2.7. Mutation in TAG lipases
• 1.2.8. Increase the TAG in vegetative tissue
• 1.3. Conclusion and future perspective
• CHAPTER 2 C-to-G base editing enhances oleic acid production by
• generating novel alleles of FATTY ACID DESATURASE 2 in plants
• 2.1. Abstract
• 2.2. Introduction
• 2.3. Materials and methods
• 2.3.1. Plant materials and growth conditions
• 2.3.2. Plasmid construction
• 2.3.3. Arabidopsis transformation and transgenic plant selection
• 2.3.4. Sanger sequencing
• 2.3.5. Fatty acid analysis
• 2.3.6. Germination rate
• 2.3.7. Measurement of root growth
• 2.3.8. Statistical analysis of the data
• 2.4. Results
• 2.4.1. Application of the base editors in planta to generate “attenuated” fad2 alleles
• 2.4.2. Analyses of the base-editing patterns and lipid contents from the fad2 lineages
• 2.4.3. Isolation of the base-edited fad2 alleles
• 2.4.4. Characterization of the growth responses of fad2 alleles
• 2.5. Discussion
• CHAPTER 3 Modulation of fatty acid elongation enhances hydroxy fatty
• acid biosynthesis and accumulation in Arabidopsis
• 3.1. Abstract
• 3.2. Introduction
• 3.3. Materials and methods
• 3.3.1. Plant materials, growth condition, and Arabidopsis transformation
• 3.3.2. Vector construction of PfKCS18
• 3.3.3. Fatty acid analysis
• 3.3.4. Analysis of the seed size, weight, and plant phenotype
• 3.3.5. Measure of the seed germination rate and the median germination time
• 3.3.6. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis
• 3.4. Results
• 3.4.1. Changes of the fatty acid composition in the pCam5, pCam5+PfKCS18, and
• pCam5-atfae1+PfKCS18 lines in the T1, T2 and T3 generations
• 3.4.2. The pCam5-atfae1+PfKCS18 increased the HFA FAME but reduced the
• total FAME content
• 3.4.3. Seed phenotype was partially recovered in pCam5+PfKCS18 and pCam5-
• atfae1+PfKCS18
• 3.4.4. Seed germination and plant growth phenotype of the transgenic lines
• 3.4.5. Expression of castor transgenes and Arabidopsis endogenous genes in the transgenic lines
• 3.5. Discussion
• CHAPTER 4 Push, pull, and protect strategy for increasing hydroxy fatty acid in Arabidopsis and Camelina sativa
• 4.1. Abstract
• 4.2. Introduction
• 4.3. Materials and methods
• 4.3.1. Arabidopsis materials, growth condition, and transformation
• 4.3.2. Camelina materials, growth condition, and transformation
• 4.3.3. Vector construction of castor genes to increase the HFA in Arabidopsis
• 4.3.4. Vector construction of castor genes to increase the HFA in camelina
• 4.3.5. Fatty acid analysis
• 4.3.6. Analysis of the seed size and weight in plants
• 4.3.7. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR)
• analysis
• 4.3.8. Statistical analysis
• 4.4. Results
• 4.4.1. Test of the push, pull, and protect strategy in accumulating HFA transgenic Arabidopsis
• 4.4.2. Seed size and dry weight-based content of the seed oil in single gene transgenic plants
• 4.4.3. Transgene and FAE1 expression in transgenic plants confirmed by RT-qPCR
• 4.4.4. Overexpression of multiple RcGPAT9, RcOLE2, and RcLPAT2 does not enhance the HFA content in the pCam5-atfae1-RcWRI1 line
• 4.4.5. Applying the strategy of enhancing HFAs by expressing five castor genes, which is proven in Arabidopsis, to the oil crop Camelina sativa
• 4.4.6. Additional introduction of castor genes into CspCam5 camelina for increasing HFAs
• 4.4.7. Seed size, weight, and dry weight-based content of the seed oil of transgenic camelina
• 4.5. Discussions
• REFERENCES
• 국문초록
• CURRICULUM VITAE