声明
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 STERILITY 1 (DPS1) encodes a CBS domain containing protein required for anther cuticle and panicle development in rice
Summary
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 qRT-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
Summary
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
参考文献
APPENDIX
List of Figures
致谢
CURRICULUM VITAE
中国农业科学院;
