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有色稻颖果色素积累的生理生态特点及其与逆境胁迫响应间关系

发布时间:2023-02-21 15:13
  水稻(Oryza sativa L.)是世界三大粮食作物之一,全球近一半人口以水稻为食。有色稻是颖果带有黑色、紫色、红色等色泽的栽培稻,是稻种资源中具有特色的栽培稻类型,其糙米含有较丰富的天然水溶性色素和矿物质等营养成分。水稻颖果的颜色变化主要受到品种遗传特性控制,但环境生态因素对有色稻颖果组织的色素积累量有明显影响。本文以籼稻9311的黑米突变体(bk)和粳稻日本晴的红米突变体(rk)及其野生型对照为材料,利用水稻籽粒灌浆期间的籽粒动态取样,对有色稻籽粒灌浆过程的色素积累动态与次生代谢相关基因表达进行了测定分析,并通过设计高温、氮素和遮阴三方面的环境因素处理,研究了环境生态因素对水稻花青素合成代谢影响及其与ROS清除抗氧化酶活性之间的相互联系。1.有色稻籽粒ANDs积累模式及其与ANDs生物合成途径相关基因表达间关系bk突变体和rk突变体籽粒中的花青素含量随籽粒灌浆而逐渐提高,成熟籽粒的花青素含量均显著高于其野生对照。其中,bk突变体籽粒中的花青素含量显著高于rk突变体,但颖果颜色突变对稻米蛋白含量和直链淀粉含量的影响不大,两个突变体与其野生型对照在籽粒蛋白质含量和直链淀粉含量上的差...

【文章页数】:189 页

【学位级别】:博士

【文章目录】:
Acknowledgement
Abstract
摘要
CHAPTER 1 Introduction and Literature review
    1.1 Economic importance of color rice
    1.2 Geographical distribution and kinds of color rice
    1.3 Types of color rice and grain pigment accumulation
    1.4 Major chemical composition and grain pigment for color rice
    1.5 Composition and structure of anthocyanidins
    1.6 Carbohydrate metabolism in color rice
        1.6.1 Total soluble sugar content
        1.6.2 Starch biosynthesis and accumulation
    1.7 Genetic control and physiological metabolism of Anthocyanin biosynthesis
        1.7.1 Anthocyanin profile of color rice
        1.7.2 Cyanidin3Glucoside (C3G)
        1.7.3 Anthocyanin Biosynthetic Pathway
        1.7.4 Genes involved in anthocyanin biosynthetic pathway
    1.8 Environmental and other Factors Affecting Pigment Accumulation
    1.9 Relationship between anthocyanin formation and defense mechanism
    Objectives of our study
CHAPTER 2 Differential anthocyanidin accumulation pattern in color rice and its relation to transcriptional expression of genes involved in anthocyanin biosynthesis pathway
    2.1 INTRODUCTION
    2.2 MATERIALS AND METHODS
        2.2.1 Plant materials and field experiment
        2.2.2 Measurements of soluble sugar content and starch accumulation in filling grains
        2.2.3 Determination of starch gelatinization characteristics
        2.2.4 Determination of various pigment contents in filling grain
        2.2.5 Measurement of anthocyanin composition by HPLC
        2.2.6 RNA isolation, cDNA preparation, and Real-time fluorescence quantitative PCR
        2.2.7 Statistical analysis
    2.3 RESULTS
        2.3.1 Mutation phenotype in the bk and rk mutant and its alteration in grain quality-related physiological parameters in mature rice grain
        2.3.2 Genotype-dependent alteration in the accumulating amount of various pigments and its temporal pattern during grain filling
        2.3.3 Genotype-dependent differences in the transcriptional profile of various genes involved in anthocyanin biosynthesis
        2.3.4 Genotype-dependent differences in the transcriptional profile and the temporal patterns of several genes involved in anthocyanin biosynthesis during filling grain
        2.3.5 Genotype-dependent difference in grain weight, soluble sugar content and starch accumulation during grain development
    2.4 DISCUSSION
    2.5 CONCLUSION
CHAPTER 3 Anthocyanidin accumulation in black kernel mutant rice and its contribution to ROS detoxification in response to high temperature at filling stage
    3.1 INTRODUCTION
    3.2 MATERIALS AND METHODS
        3.2.1 Plant materials and temperature treatments
        3.2.2 Examination of grain ultrastructure
        3.2.3 Determination of anthocyanidin, flavonoid and polyphenol contents in rice grains
        3.2.4 Measurement of anthocyanins composition by HPLC
        3.2.5 RNA isolation, cDNA preparation, and Real-time fluorescence quantitative PCR
        3.2.6 Determination of malondialdehyde (MDA), hydrogen peroxide (H2O2), superoxide radical (O2
-), and antioxidant enzyme activity
        3.2.7 Determination of carbohydrates
        3.2.8 Statistical analysis
    3.3 RESULTS
        3.3.1 Differences in kernel pigment concentration and ANDs composition between 9311bk and wild type under different temperature regimes
        3.3.2 Expression pattern of various genes involved in ANDs biosynthesis in developing kernels and its response to HT exposure at grain filling
        3.3.3 Difference in the response of kernel ROS content, MDA accumulation, antioxidant enzyme, to HT exposure between bk mutant rice and its wild type
        3.3.4 Ultrastructural changes induced by HT in kernel of bk mutant and its wild type
    3.4 DISCUSSION
        3.4.1 ANDs deficiency in wild type confers more endogenous ROS accumulation under high-temperature stress
        3.4.2 ANDs accumulation confers more ROS scavenging ability in bk mutant than wild type under high-temperature stress
    3.5 Conclusion
CHAPTER 4 Effect of nitrogen deficiency at filling stage to kernel anthocyanidin accumulation and its relation to ROS detoxification in filling grains for bk mutant
    4.1 INTRODUCTION
    4.2 MATERIALS AND METHODS
        4.2.1 Plant materials and treatments
        4.2.2 Determination of anthocyanidin, flavonoid and polyphenol contents in rice grains
        4.2.3 RNA isolation, cDNA preparation, and Real-time fluorescence quantitative PCR
        4.2.4 Determination of malondialdehyde (MDA), hydrogen peroxide (H2O2) and superoxide radical (O2
-), total soluble sugar and sucrose contents in rice grains
        4.2.5 Determination of antioxidant enzyme activity
        4.2.6 Statistical analysis
    4.3 RESULTS
        4.3.1 Effect of N deficiency at filling stage on the protein and pigments concentration and anthocyanidin composition in bk mutant grains
        4.3.2 Change in the transcriptional expression of various genes involved in anthocyanin biosynthesis in rice for the bk mutant under N-deficiency
        4.3.3 Difference in the response of kernel ROS content, MDA accumulation, antioxidant enzyme to N deficiency in bk mutant rice
    4.4 DISCUSSION
    4.5 Conclusion
CHAPTER 5 Effect of shading at filling stage to kernel anthocyanidin accumulation and its relation to ROS detoxification in filling grains for bk mutant
    5.1 INTRODUCTION
    5.2 MATERIALS AND METHODS
        5.2.1 Plant materials and treatments
        5.2.2 Examination of grain ultrastructure
        5.2.3 Determination of anthocyanidin, flavonoid and polyphenol contents in rice grains
        5.2.4 Measurement of anthocyanins composition by HPLC
        5.2.5 RNA isolation, cDNA preparation, and Real-time fluorescence quantitative PCR
        5.2.6 Determination of malondialdehyde (MDA), hydrogen peroxide (H2O2) and superoxide radical (O2
-), total soluble sugar and sucrose contents in rice grains
        5.2.7 Determination of antioxidant enzyme activity
        5.2.8 Statistical analysis
    5.3 RESULTS
        5.3.1 Effect of shading at filling stage on the pigment concentration and anthocyanidin composition in bk mutant grains
        5.3.2 Change in the transcriptional expression of various genes involved in anthocyanin biosynthesis in rice for the bk mutant and its wild type under shading condition
        5.3.3 Difference in the response of kernel ROS content, MDA accumulation and antioxidant enzyme activity to shading condition in bk mutant and its wild type
        5.3.4 Difference in the response of kernel weight, soluble sugar, sucrose and starch content to shading in bk mutant
        5.3.5 Ultrastructural changes induced by HT in kernel of bk mutant and its wild type
    5.4 DISCUSSION
    5.5 Conclusion
CHAPTER 6 MAJOR FINDINGS AND FUTURE PERSPECTIVES
    6.1 Major findings
    6.2 Schematic diagram of research summary
    6.3 Future perspectives
References
LIST OF SCIENTIFIC PUBLICATIONS



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