水稻DPS1基因調(diào)控穗部和葉片發(fā)育的研究
發(fā)布時(shí)間：2023-04-05 19:23
為供養(yǎng)不斷膨脹的世界人口,提高糧食生產(chǎn)力變得勢(shì)在必行。世界上一半以上的人口以水稻為主食。水稻產(chǎn)量取決于穗大小、穗粒數(shù)、穗育性和結(jié)實(shí)率等性狀。逆境條件如冷或熱脅迫會(huì)導(dǎo)致水稻穗部退化(敗育)和不育,從而導(dǎo)致水稻籽粒產(chǎn)量下降�；ㄋ幨侵参镄坌陨称鞴�,被角質(zhì)層所覆蓋保護(hù),以避免花藥水分流失。然而,花藥角質(zhì)層發(fā)育和穗退化的分子機(jī)制仍有待探索。本文研究了一個(gè)新的水稻育性突變體degenerated panicle and partial sterility 1(dps1),該突變體表現(xiàn)為花序頂端退化、生育力下降以及旗葉扭曲并產(chǎn)生黃色斑塊。主要研究結(jié)果如下:i)dps1突變體的表型分析顯示其花序頂端小穗敗育,花藥白色,無花粉粒。ii)dps1突變體中約60%的小穗不育,導(dǎo)致dps1的結(jié)實(shí)率(38±5.2%)明顯低于WT(95±2%)。iii)掃描電鏡顯示dps1花藥的角質(zhì)層形態(tài)與WT相比發(fā)生了改變,dps1花藥的角質(zhì)層蠟質(zhì)和角質(zhì)含量明顯減少。iv)TUNEL和Comet檢測(cè)顯示程序性細(xì)胞死亡(PCD)增加,導(dǎo)致dps1花序和花藥的DNA片段化增強(qiáng)。v)與野生型相比,dps1穗部活性氧(ROS)、丙二...
【文章頁(yè)數(shù)】：88 頁(yè)
【學(xué)位級(jí)別】：博士
【文章目錄】：
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
本文編號(hào)：3783982
【文章頁(yè)數(shù)】：88 頁(yè)
【學(xué)位級(jí)別】：博士
【文章目錄】：
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
本文編號(hào)：3783982
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