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水楊酸和谷胱甘肽調(diào)控鉻脅迫下油菜不同耐性品種生理生化和基因組變化的作用機(jī)理

發(fā)布時(shí)間:2021-01-18 12:49
  土壤中重金屬的毒害是制約油菜產(chǎn)量的主要因素之一。篩選對(duì)重金屬抗性高的基因型是最佳解決方法之一,使用不同的植物生長(zhǎng)調(diào)節(jié)劑提高非生物脅迫下長(zhǎng)勢(shì)也有顯著效果。本研究包括四部分試驗(yàn)內(nèi)容,取得結(jié)果如下:(1)試驗(yàn)以四個(gè)不同甘藍(lán)型油菜品種(浙雙758、浙大619、浙油50、浙大622)的6天幼苗為試驗(yàn)材料,研究鉻(cr)的毒害作用。結(jié)果表明,隨著鉻濃度的升高,所有品種的幼苗生長(zhǎng)均受到抑制,光合色素含量和抗氧化酶活性顯著降低,丙二醛(MDA)和活性氧(ROS)的含量上升。相對(duì)于其他品種,浙大622的各個(gè)部位的鉻含量均較高。電鏡結(jié)果表明,400μM鉻處理下,葉肉和根尖細(xì)胞超微結(jié)構(gòu)被完全破壞,浙大622的破壞效果最顯著。因此,浙大622相對(duì)于其他3個(gè)油菜品種表現(xiàn)更敏感。(2)選取了水培生長(zhǎng)下的四個(gè)不同甘藍(lán)型油菜品種來(lái)研究鉻的毒性。結(jié)果表明,隨著鉻濃度的升高,油菜的生物量有所降低。而浙大622下降幅度最為顯著,其光合作用降低,而浙雙758在鉻的毒害下相對(duì)表現(xiàn)較好。不同濃度的鉻誘導(dǎo)了活性氧和丙二醛的積累。然而,為了清除活性氧和丙二醛的積累,相關(guān)的酶活性在葉片和根中均升高。電鏡結(jié)果表明,400μM鉻處理下浙大... 

【文章來(lái)源】:浙江大學(xué)浙江省 211工程院校 985工程院校 教育部直屬院校

【文章頁(yè)數(shù)】:160 頁(yè)

【學(xué)位級(jí)別】:博士

【文章目錄】:
Thesis Release Certificate
Certification
ACKNOWLEDGEMENTS
LIST OF TABLES
LIST OF FIGURES
ABBREVIATIONS
Abstract
摘要
CHAPTER 1
    1.1 What is oilseed rape
    1.2 Economic importance
    1.3.Promising heavy metals and their concentration in the soil of ZhejiangProvince
    1.4 Interaction of chromium and plants
    1.5. Interaction of Cr and plants
    1.6. Possible way outs
    1.7. Aims of the study
    1.8. Study outlines
CHAPTER 2
    2.1 What is stress
    2.2 Cr as a toxic and non essential element
        2.2.1 Occurrence
        2.2.2 Cr species
    2.3 Human health and Cr toxicity
    2.4 Hazardous effects of Cr on plants
    2.5 Cr and Brassica napus interaction
    2.6 SA as an elevator
    2.7 Role of GSH to improve the stress impacts on plants
CHAPTER 3
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Plant material and experimental conditions
        3.2.2 Measurement of Plant physio-morphic attributes
        3.2.3 Determination of MDA and singlet oxygen species
        3.2.4 Assessment of enzyme based plant immune system
        3.2.5 Ultrastructural observations
        3.2.6 Statistical analysis
    3.3 Results
        3.3.1 Analyses of physiological parameters
        3.3.2 Determination of Cr and chlorophyll contents
        3.3.3 MDA and ROS conten estimation
        3.3.4 Evaluation of defense related enzymes and TSP contents
        3.3.5 TEM ultrastructural observations of leaf
        3.3.6 TEM ultrastructural observations of root
    3.4 Discussion
    3.5 Conclusions
CHAPTER 4
    4.1 Introduction
    4.2 Materials and methods
        4.2.1 Plant material and Growth conditions
        4.2.2 Morphological parameters
        4.2.3 Analysis of lipid peroxidation and reactive oxygen species(ROS)
        4.2.4 Biochemical analysis
        4.2.5 Analyses of detoxification related antioxidants
        4.2.6 Transmission electron microscopy
        4.2.7 Statistical analysis
    4.3 Results
        4.3.1. Morphological attributes
        4.3.2 Photosynthetic gas exchange capacity
        4.3.3. Estimation of MDA and ROS
        4.3.4. Analysis of enzymatic antioxidants and TSP
        4.3.5. Analysis of detoxification related defense mechanism
        4.3.6. TEM cell structural observations
    4.4 Discussion
    4.5 Conclusions
CHAPTER 5
    5.1 Introduction
    5.2 Materials and methods
        5.2.1 Plant material and growth conditions
        5.2.2 Chlorophyll and Cr content determination
        5.2.3 Extraction and estimation of total SA contents
        5.2.4 Determination of MDA contents and ROS activity
        5.2.5 Determin ation of antioxidant machinery
        5.2.6 Total RNA extraction,cDNA synthesis,and quantitative real-time PCR assay
        5.2.7 Ultrastructural observations
        5.2.8 Statistical analysis
    5.3 Results
        5.3.1. Morpho-physiological attributes
        5.3.2. Lipid peroxidation and ROS determination
        5.3.3. Antioxidants enzyme activities
        5.3.4. Enzymatic antioxidants transcript level
        5.3.5. Chromium and total salicylic acid contents
        5.3.6. Detoxification and secondary metabolite related gene expressions
        5.3.7. Cell ultra-structural observations
    5.4 Discussion
    5.5 Conclusions
CHAPTER 6
    6.1 Introduction
    6.2 Materials and methods
        6.2.1 Plant material and growth conditions
        6.2.2. Cr and GSH content determination
        6.2.3 Total RNA extraction,reliability assessment and RNA-sequence analyses
        6.2.4 Establishment of de-novo assembly
        6.2.5. Screening of differentially expressed genes(DEGs) and group differentially expressed genes
        6.2.6. Expression pattern analysis of DEGs
        6.2.7. Gene Ontology(GO)functional enrichment analysis(WEGO)of DEGs
        6.2.8. KEGG pathway enrichment analysis
        6.2.9. Satistical analysis
    6.3 Results
        6.3.1. Plant biomass,Cr and GSH contents determination
        6.3.2. RNA sequencing and denovo assembly
        6.3.3. Screening,expression pattern analysis and clustering of differentially expressed genes(DEGs)
        6.3.4. Gene ontology functional classification(WEGO)of DEGs and cluster of orthologous groups(COGs)classification
        6.3.5. KEGG metabolic pathway enrichment analysis
        6.3.6. Analysis of top 10 up-regulated stress response-related DEGs
    6.4 Discussion
    6.5 Conclusions
CHAPTER 7
    7.1 Major findings
    7.2 Future perspectives
References
List of Publications



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