High-throughput screening to explore novel allelic variations of GA20ox1 via CRISPR-mediated adenine base editing in Arabidopsis

다국어 초록 (Multilingual Abstract)

Gibberellin 20-oxidase 1 (GA20ox1), one of the 'green revolution’ genes, is an enzyme involved in the biosynthesis of gibberellins, which regulates plant growth in diverse developmental stages. Natural alleles found in GA20ox1 were proposed as the causal locus of semi-dwarfism observed in Arabidopsis ecotypes, but no functional validation has been tested so far. In the previous study, allelic variations of GA20ox1 were obtained while reproducing Mar-1 ecotype via CRISPR-mediated cytosine base editor (CBE). Here, I confirmed the association between GA20ox1 allelic variations and plant growth patterns. Subsequently, I extensively employed a high-throughput strategy using CRISPR- mediated adenine base editor (ABE), which enabled the identification of novel allelic variations of GA20ox1 linked to plant growth traits. The novel findings and high-throughput strategy can provide critical insights to the functional study of genes and the research on crop improvement.

목차 (Table of Contents)

• Ⅰ. LIST OF TABLES ⅲ
• Ⅱ. LIST OF FIGURES ⅳ
• Ⅲ. ABSTRACT ⅵ
• Ⅳ. CHAPTER 1 1
• 1. A. INTRODUCTION 2
• 1. B. MATERIALS AND METHODS 11
• 1. C. RESULTS 17
• 1. D. DISCUSSION 33
• Ⅴ. CHAPTER 2 36
• 2. A. INTRODUCTION 37
• 2. B. MATERIALS AND METHODS 44
• 2. C. RESULTS 56
• 2. D. DISCUSSION 67
• Ⅵ. REFERENCES 69
• Ⅶ. APPENDICES 74