棉花是重要的經(jīng)濟(jì)作物,隨著全球氣溫的逐年升高,棉花的產(chǎn)量受到高溫脅迫的威脅越來越嚴(yán)重。高溫脅迫對植物生殖器官的影響要遠(yuǎn)大于營養(yǎng)器官,且雄性生殖器官對溫度的敏感程度高于雌性生殖器官,所以高溫脅迫下的雄性不育更應(yīng)受到關(guān)注。因不同的高溫類型會引起不同的雄性不育表型,為了分析不同高溫類型導(dǎo)致雄性敗育發(fā)生的機(jī)理,我們設(shè)置了四種溫度組合條件（白天常溫/晚間常溫,NN;白天高溫/晚間常溫,HN;白天常溫/晚間高溫,NH;白天高溫/晚間高溫,HH）進(jìn)行處理,并對處理后的棉花三個發(fā)育時期（四分體時期、絨氈層降解時期、花藥開裂期）的花藥進(jìn)行組織細(xì)胞水平,生理生化水平及轉(zhuǎn)錄水平分析。研究發(fā)現(xiàn),高夜溫對于棉花雄性育性的影響要遠(yuǎn)遠(yuǎn)大于高日溫,其造成的雄性不育是引起棉花減產(chǎn)的主要原因之一。具體研究結(jié)果如下:1)高溫會導(dǎo)致花器官明顯變小,花絲變短,花粉不育,其中全天高溫（HH）造成的不育最明顯,且高夜溫（NH）引起的花粉不育比高日溫（HN）要明顯。通過花藥切片觀察發(fā)現(xiàn),在絨氈層降解時期,全天高溫條件下的小孢子出現(xiàn)明顯皺縮,而在白天高溫/晚間正常和白天正常/晚間高溫條件下有部分小孢子出現(xiàn)皺縮,程度要輕于全天高溫。2)... 

【文章來源】：華中農(nóng)業(yè)大學(xué)湖北省 211工程院校 教育部直屬院校

【文章頁數(shù)】：136 頁

【學(xué)位級別】：博士

【文章目錄】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER I
    1 INTRODUCTION
        1.1 High Temperature（HT）stress
        1.2 High temperature stress and its effect on crops
        1.3 High temperature stress and its effect on Cotton
        1.4 High temperature and flowering time
        1.5 High temperature and fertility
        1.6 High temperature affects pollen development
        1.7 High temperature and its effect on carbohydrate
        1.8 High temperature and its effects on lipids
        1.9 High temperature and its effect on ROS in the anthers
        1.10 High temperature and hormones
            1.10.1 Auxin and male sterility
            1.10.2 Gibberellin（GA）and male sterility
            1.10.3 Jasmonic Acid（JA）and male sterility
        1.11 CRISPR/Cas9 system in plants
            1.11.1 Mechanism of CRISPR-Cas9 system
            1.11.2 Applications of CRISPR/Cas9 in Crop improvement
        1.12 Aims and overview of the present study
CHAPTER II
    2.MATERIALS AND METHODS
        2.1 Plant material
        2.2 Pollen viability,Tissue sectioning and observation
        2.3 RNA sequencing
        2.4 Data analysis
        2.5 Quantitative Reverse Transcriptase PCR（q RT-PCR）
        2.6 Fatty acid extraction and measurement
        2.7 Transmission electron microscopy（TEM）
        2.8 Measurement of ABA,IAA,and JA
        2.9 Quantification of H2O2
        2.10 Starch,sucrose and fructose measurement
        2.11 Vector construction and transformation in cotton
            2.11.1 PCR conditions:
            2.11.2 Second PCR and purification
            2.11.3 PCR conditions:
            2.11.4 Enzyme digestion of the expression vector:
            2.11.5 In-fusion ligation
            2.11.6 Heat shock and electroporation
        2.12 Plant material,growth conditions,and treatments
        2.13 Tissue culturing and generation of homozygous transgenic lines
        2.14 DNA extraction and Southern blotting
        2.15 Hi-TOM sequencing
        2.16 Application of exogenous Me JA to buds
        2.17 Reactive oxygen species（ROS）staining
        2.18 Statistical analysis
CHAPTER III
    3.RESULTS
        3.1 Phenotypic studies of the high temperature（HT）sensitive line H05 under day and night HT stress
        3.2 High night temperature causes pollen abortion
        3.3 Differentially expressed genes（DEGs）in H05 anthers under different HT stress combinations
        3.4 HT stress disrupts cotton anthers lipid metabolism pathways
        3.5 Day and night HT affect fatty acid metabolism differently
        3.6 Day and night HT drive Alpha-linolenic acid metabolism to JA in ADS stage
        3.7 Effects of HT on the glycerolipid metabolism pathway genes
        3.8 Day and night HT affects the lipid and sugar contents in anthers
        3.9 Study of selected genes from the Jasmonic Acid（JA）pathway
            3.9.1 Structural characterization of Allene oxide cyclase4（AOC4）and Acyl-Co A oxidase2（ACO2）
            3.9.2 Transformation and plant regeneration
            3.9.3 Selection of Gh AOC4 and Gh ACO2 mutant lines
            3.9.4 Phenotypic study and histological sectioning of Gh AOC4 and Gh ACO2 mutant lines anthers
            3.9.5 Transmission electron microscopy（TEM）of the mutant lines anthers
            3.9.6 Gh AOC4 and Gh ACO2 gene disrupts the JA pathway and caused pollen abortion
            3.9.7 High temperature（HT）decreases JA accumulation in the anthers of cotton HT-sensitive line anthers and causes male sterility
            3.9.8 Decreased JA biosynthesis causes the accumulation of H2O2 in the anther and pollen of the Gh AOC4 mutant
CHAPTER IV
    4.DISCUSSION
        4.1 Lipid and fatty acid changes are associated with male sterility under both day and night HT stress
        4.2 Day and Night HT disrupt the glycerolipid and JA pathway genes,leading to male sterility
        4.3 Disruption of hormones during day and night HT leads to male sterility
        4.4 Knock out of Gh AOC4 and Gh ACO2 leads to male sterility
        4.5 Gh ACO2 and Gh AOC4 are responsible for the sterile pollen and anther indehiscence
CONCLUSION
REFERENCES
APPENDICES
    APPENDIX 1
    APPENDIX 2
    APPENDIX 3
    APPENDIX 4
    APPENDIX 5
    APPENDIX 6
    APPENDIX 7
    APPENDIX 8
ACKNOWLEDGMENT
List of Publications



本文編號：3066187