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研究生: 阮明可
Nguyen, Minh Khiem
論文名稱: 水稻葉綠素缺失突變株之葉綠素新陳代謝之研究
Chlorophyll Metabolism of Chlorophyll-Deficient Mutant of Rice (Oryza sativa L.)
指導教授: 楊棋明
Yang, Chi-Ming
鍾國芳
Chung, Kuo-Fang
口試委員: 林冠宏
Lin, Kuan-Hung
黃盟元
Huang, Meng-Yuan
孫智雯
Sun, Chih-Wen
傅瀚儀
Fu, Han-Yi
口試日期: 2021/06/04
學位類別: 博士
Doctor
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2021
畢業學年度: 109
語文別: 英文
論文頁數: 154
英文關鍵詞: Chl-deficient mutant, Grana, Photosynthesis, Mutant, Oryza sativa
研究方法: Pigment assessmentUltra structrure analysisNext Generation SequencingRT-qPCRRNA-Seq
DOI URL: http://doi.org/10.6345/NTNU202100554
論文種類: 學術論文
相關次數: 點閱:100下載:3
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  • Photosynthesis is a vital biological process in higher plants, which contributes to approximately 90% of biomass production in crops. Therefore, enhancing photosynthesis is considered as an effective strategy for increasing crop yield. In addition, photosynthesis take place in chloroplasts where chlorophylls (Chl) biosynthesis and degradation occur. Chloroplast pigments are extremely important during photosynthesis since they play an essential role in light absorption and energy transfer. Therefore, understanding and enhancing the efficiency of Chl biosynthesis are a potential way to increase accumulation of pigments and crop yield.
    Chl-deficient mutants is valuable approach for studying pigment synthesis. Generally, Chl-deficient mutants can be classified into two main types, namely, Chl b-lacking mutants (with undetectable Chl b) and Chl b-deficient mutants (containing reduced levels of Chl b). Next-generation sequencing (NGS) for transcriptome profiling was used to determine the transcription profiles and photosynthetic characteristics underlying different Chl a/b ratio of Chl b-lacking rice (Chl a/b ratio of ch1); Chl b-deficient rice (Chl a/b ratio of 11.25, ch11) and type 2b (Chl a/b ratio of 15.7, ch14).
    In the first chapter, the literature on the characteristic of chlorophyll-deficient mutants and the function and metabolism of Chl in higher plant.
    In the second, third and fourth chapter, the photosynthetic properties and transcriptomic profiles of wildtype and Chl b-lacking (ch11), 2 types of Chl b deficient mutants (ch11, ch14) of rice were investigated. Those mutants exhibited dwarf phenotype, light green leaves, and abnormal chloroplast structure (i.e., loss or small of starch granules, abundant vesicles, and abundant plastoglobuli), indicating abnormal plastid development with amplifier Chl a/b ratio. Changes in the expression of genes related to Chl metabolism, chloroplast development, cell division, and photosynthesis were found to be associated with abnormal chloroplast development and reduced Chl accumulation in the mutants. qPCR analysis was used to validate the DEGs. The data indicated that an increase in the Chl a/b may attribute to both a reduction in Chl content, owing to abnormal chloroplast development, and the involvement of an alternative degradation pathway.
    In the fifth chapter, the effect of temperature on the Chl biosynthesis and characteristic of ch1 rice was further investigated. The data indicated that ch1 was sensitive to low temperature at beginning of incubation and may adapt to temperatures ranging from 15 °C to 35 °C.
    In the sixth chapter, the effect of shade conditions on Chl biosynthesis in ch1 and wt was evaluated. Pigment contents, ultrastructure analysis, and RT-qPCR was evaluated in ch1 under 2 shade conditions. The data revealed that Chl b was unable to be generated in ch1 rice whereas Chl a remained unchanged. The results suggested that Chl b in ch1 rice was degraded or unable to generated rather than rapidly converted to Chl a during shade conditions.
    The final chapter discussed the finding and future works on Chl-deficient mutants.

    TABLE OF CONTENTS ACKNOWLEDGEMENTS i ABSTRACT ii LIST OF FIGURES vi LIST OF TABLES x ABBREVIATIONS xi CHAPTER 1 . 1 Literature review 1 Background of Chlorophyll-Deficient Mutants 2 Photosynthesis 2 Chlorophyll Biosynthesis and Degradation Pathway 3 References 7 CHAPTER 2 . 12 Transcription Analysis of Chlorophyll Biosynthesis in Wild-type and Chlorophyll b-Lacking Rice (Oryza sativa L.) 12 Abstract 13 Introduction 14 Materials and Methods 15 Results 16 Discussion 18 Conclusion 22 Supplementary material 32 References 36 CHAPTER 3 . 41 Transcription profile analysis of chlorophyll biosynthesis in leaves of wild-type and chlorophyll b-deficient rice (Oryza sativa L.) 41 Abstract 42 Introduction 43 Materials and Methods 45 Results 47 Discussion 49 Conclusions 54 References 69 CHAPTER 4 . 76 Chlorophyll biosynthesis and transcriptome profiles of chlorophyll b-deficient type 2b rice (Oryza sativa L.) 76 Abstract 77 Introduction 78 Materials and Methods 79 Results 81 Discussion 83 Conclusions 87 References 101 CHAPTER 5 . 107 Temperature Mediated Shifts in Chlorophyll Biosynthesis in Leaves of Chlorophyll b-Lacking Rice Leaves (Oryza sativa L.) 107 Abstract 108 Introduction 109 Materials and Methods 110 Results 112 Discussion 113 Conclusions 116 References 124 CHAPTER 6 . 131 Chlorophyll Biosynthesis and Relative Gene Expression in Chlorophyll b-Lacking Rice (Oryza sativa L.) at Low Light Intensities 131 Abstract 132 Introduction 133 Materials and Methods 134 Results 136 Discussion 137 Conclusions 140 References 147 CHAPTER 7 . 152 Conclusion and Future Works 152

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