水稻DPS1基因调控穗部和叶片发育的研究

发布时间：2023-04-05 19:23

　　为供养不断膨胀的世界人口,提高粮食生产力变得势在必行。世界上一半以上的人口以水稻为主食。水稻产量取决于穗大小、穗粒数、穗育性和结实率等性状。逆境条件如冷或热胁迫会导致水稻穗部退化(败育)和不育,从而导致水稻籽粒产量下降。花药是植物雄性生殖器官,被角质层所覆盖保护,以避免花药水分流失。然而,花药角质层发育和穗退化的分子机制仍有待探索。本文研究了一个新的水稻育性突变体degenerated panicle and partial sterility 1(dps1),该突变体表现为花序顶端退化、生育力下降以及旗叶扭曲并产生黄色斑块。主要研究结果如下:i)dps1突变体的表型分析显示其花序顶端小穗败育,花药白色,无花粉粒。ii)dps1突变体中约60%的小穗不育,导致dps1的结实率(38±5.2%)明显低于WT(95±2%)。iii)扫描电镜显示dps1花药的角质层形态与WT相比发生了改变,dps1花药的角质层蜡质和角质含量明显减少。iv)TUNEL和Comet检测显示程序性细胞死亡(PCD)增加,导致dps1花序和花药的DNA片段化增强。v)与野生型相比,dps1穗部活性氧(ROS)、丙二...

【文章页数】：88 页

【学位级别】：博士

【文章目录】：
摘要
Abstract
List of Abbreviations
CHAPTER Ⅰ–BACKGROUND
    1.1 Panicle development in rice
    1.2 Panicle degeneration in rice
    1.3 Panicle fertility and anther development in rice
    1.4 Leaf chloroplast and cuticle development
    1.5 Role of ROS in plant development and reproduction
    1.6 Cystathionine-β-synthase(CBS)domain-containing proteins
    1.7 Objectives of the study
CHAPTER Ⅱ- DEGENERATED PANICLE AND PARTIAL STERILITY1 (DPS1)ENCODES A CBS DOMAIN CONTAINING PROTEIN REQUIRED FOR ANTHER CUTICLE AND PANICLE DEVELOPMENT IN RICE
    2.1 INTRODUCTION
    2.2 MATERIALS AND METHODS
        2.2.1 Plant materials and growth conditions
        2.2.2 Phenotypic characterization
        2.2.3 Scanning electron microscopy(SEM)
        2.2.4 Analysis of anther wax and cutin
        2.2.5 TUNEL assay for nuclear DNA fragmentation
        2.2.6 Comet assay for DNA damage
        2.2.7 Staining for ROS
        2.2.8 Quantitative measurement of ROS
        2.2.9 MDA content and antioxidants activities
        2.2.10 Map-based cloning
        2.2.11 Vector construction and transformation
        2.2.12 DPS1-promoter-GUS assay
        2.2.13 Subcellular localization of DPS1
        2.2.14 Yeast two-hybrid assay
        2.2.15 Pull-down assay
        2.2.16 Luciferase imaging assay
        2.2.17 Transcriptome analysis
        2.2.18 RNA isolation and q RT-PCR
        2.2.19 Phylogenetic analysis
        2.2.20 Accession numbers
    2.3 RESULTS
        2.3.1 The dps1 mutant exhibits degenerated apical spikelets and sterile middle spikelets
        2.3.2 The middle spikelets of dps1 are pollenless and defective in anther cuticle development
        2.3.3 Amounts of wax and cutin in dps1 anthers are reduced
        2.3.4 dps1 mutant displays enhanced cell death
        2.3.5 DPS1 encodes a CBSDUF protein
        2.3.6 DPS1 gene is preferentially expressed during late reproductive development and DPS1 protein is localized to mitochondria
        2.3.7 dps1 accumulates more ROS in defective panicles
        2.3.8 DPS1 interacts with thioredoxin proteins Trx1 and Trx20
        2.3.9 Transcriptome analysis highlights role of DPS1-mediated ROS level in regulating fatty acid metabolism
        2.3.10 Exogenous H2O2 application affects the expression of cuticle associated genes
    2.4 DISCUSSION
        2.4.1 DPS1 function is associated with seed setting in rice
        2.4.2 DPS1 is involved in anther cuticle formation
        2.4.3 DPS1 is required for maintaining ROS homeostasis
        2.4.4 DPS1 interacts with thioredoxin proteins to regulate ROS homeostasis
CHAPTER Ⅲ- DPS1 REGULATES LEAF CHLOROPLAST AND CUTICLE DEVELOPMENT IN RICE BY ROS-MEDIATED CELL DEATH
    3.1 INTRODUCTION
    3.2 MATERIAL AND METHODS
        3.2.1 Plant materials and growth conditions
        3.2.2 Pigment analysis
        3.2.3 Transmission electron microscopy
        3.2.4 Scanning electron microscopy
        3.2.5 Chlorophyll leaching assay
        3.2.6 Toluidine blue staining
        3.2.7 Quantification of wax and cutin
        3.2.8 Vector construction and transformation
        3.2.9 TUNEL assay
        3.2.10 Histochemical staining and quantitative measurement of ROS
        3.2.11 Measurement of MDA
    3.3 RESULTS
        3.3.1 DPS1 regulates chloroplast development and leaf shape
        3.3.2 DPS1 control leaf cuticle development
        3.3.3 Genetic complementation using DPS1
        3.3.4 dps1 mutant leaves had enhanced cell death and ROS accumulation
    3.4 DISCUSSION
CHAPTER Ⅳ CONCLUSION
REFERENCES
APPENDIX
LIST OF FIGURES
ACKNOWLEDGEMENT
CURRICULUM VITAE



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