基于代谢组和遗传适合度分析高镉积累油菜中镉的适应性响应
发布时间：2024-02-02 23:33
镉(Cd)作为可致癌金属,是环境中广泛存在的有毒重金属之一。由于土壤中的Cd污染而引起的环境问题日益严重,因此鼓励采取公共卫生防治措施极为重要。甘蓝型油菜(Brassica napus L.)因与Cd超富集物种在某些特征上具有相似性,被认为是修复Cd污染土壤的最佳选择之一。然而Cd在营养和生殖阶段的解毒机理尚未完全阐明,尽管这是使该作物易于应用于植物修复技术的前提。本研究采用短期(水培)和长期(土培)的方法,旨在探究耐镉甘蓝型油菜中质外体和共质体对Cd积累耐性的反应机理,并着重于代谢组的分析,同时分析了镉对植物生殖适应性的影响。实验结果如下:(1)为了筛选Cd耐受性的油菜材料,本研究利用102个不同遗传背景的甘蓝型油菜材料,经过两次筛选,测试了代表不同Cd耐性范围的11个基因型的根和芽中Cd积累和转运。发现Cd抗性和积累的显着变化,并鉴定出两个耐高Cd积累的基因型(CB671和HL672)。巧合的是,这两个基因型是白花油菜的基因型,命名为“高镉基因型”。在102个被筛选的基因型中,只有两个具有Cd-超积累的表现,这表明该表型在甘蓝型油菜中很少发生。因此,这些材料(高镉基因型)成为后续研...
【文章页数】：159 页
【学位级别】：博士
【文章目录】：
Acknowledgements
List of abbreviations
Abstract
摘要
CHAPTER1.General Introduction and Literature Review
1.1.Cadmium occurrence in the environment
1.1.1.Worldwide occurrence of cadmium
1.1.2.Cadmium-soil contamination in China
1.2.Environmental issues of cadmium exposure
1.2.1.Cadmium phytotoxicity effects
1.2.2 Cadmium and human health risks
1.3.Brassica napus,a tool for sustainable management of Cd-contaminated soils
1.4.Current state of knowledge on Cd adaptive responses in Brassica napus
1.5.Research objectives
1.6.Outline of the study
CHAPTER2.Intraspecific variability of cadmium accumulation,subcellular distribution and chemical forms in Brassica napus
2.1.Introduction
2.2.Materials and methods
2.2.1.Plant material and experimental conditions
2.2.2.Determination of cadmium content
2.2.3.Plant tissues fractionation and metal analysis
2.2.4.Analysis of metal chemical forms
2.2.5.Statistical analysis
2.3.Results
2.3.1.Variation in cadmium-induced changes in plant biomass,and trend of metal accumulation
2.3.2.Metal subcellular distribution
2.3.3.Metal chemical forms
2.4.Discussion
2.5.Conclusion
CHAPTER3.Cadmium detoxification in high Cd-accumulating Brassica napus genotypes involves changes in cell wall compositional profiles
3.1.Introduction
3.2.Materials and methods
3.2.1.Plant material and experimental conditions
3.2.2.Cell wall extraction and polysaccharides fractionation
3.2.3.Measurement of uronic acid and total sugar in fractionated cell wall polysaccharides
3.2.4.Quantification of different fractions of pectin and estimation of its methyl-esterification degree
3.2.5.Determination of Cd contents in fractionated cell wall polysaccharides
3.2.6.Measurement of ascorbic acid content
3.2.7.Processing of cell wall-related RNA-seq data
3.2.7.1.Total RNA extraction,reliability assessment and RNA-sequence analyses
3.2.7.2.Verification of RNA-Seq data by qRT-PCR assay
3.2.8.Statistical analysis
3.3.Results
3.3.1.Cd-induced remodelling of cell wall polysaccharides among B.napus genotypes
3.3.2.Relative contribution of different cell wall sub-fractions to Cd immobilization
3.3.3.Cadmium influence on transcriptional regulation of pectin and hemicellulose biosynthesis
3.3.4.Pectin as possible substrat for ascorbic acid production in leaves
3.4.Discussion
3.5.Conclusion
CHAPTER4.Comparative metabolomic responses of low-and high-cadmium accumulating genotypes reveal the Cd adaptive mechanism in Brassica napus
4.1.Introduction
4.2.Materials and methods
4.2.1.Plant material and experimental conditions
4.2.2.Metabolomic analysis
4.2.2.1.Metabolites extraction and liquid chromatography-mass spectrometry
4.2.2.2.Data processing
4.2.3.Analysis of other physio-biochemical endpoints
4.2.3.1.Assessment of Cd accumulation in plant tissues
4.2.3.2.Transmission electron microscopy,scanning electron microscopy and chlorophyll fluorescence imaging
4.2.3.3.Determination of phenolic contents and total antioxidant capacity
4.2.3.4.Total RNA extraction,cDNA synthesis,and qRT-PCR assay
4.2.4.Statistical analyses
4.3.Results
4.3.1.Genotypic variation of responses to cadmium stress
4.3.2.General metabolomic changes in the two genotypes following Cd stress
4.3.3.Profiling of metabolites specifically regulated in either genotype under Cd stress
4.3.3.1.Metabolomic signature in plant primary metabolism
4.3.3.2.Metabolomic signature in plant secondary metabolism
4.4.Discussion
4.5.Conclusion
CHAPTER5.Cadmium-induced reproductive repercussions in two Brassica napus genotypes supplemented with serotonin,and glimpses into potential adaptive mechanisms
5.1.Introduction
5.2.Materials and methods
5.2.1.Plant material and experimental conditions
5.2.2.Data collection and samples preparation for analysis
5.2.3.Elements analysis in different plant parts
5.2.4.Seed oil content determination
5.2.5.Seed transmission electron microscopy
5.2.6.Evaluation of pollen grain viability and cross-pollination experiments
5.2.7.Measurement of other physio-biochemical end points
5.2.7.1.Determination of indices of oxidative stress
5.2.7.2.Photosynthetic parameters
5.2.7.3.Determination of antioxidant enzymes
5.2.7.4.Thiol compounds assays
5.2.7.5.Assays of activities of sulfur assimilation enzymes
5.2.7.6.Measurement of phenolic contents
5.2.8.Data analysis
5.3.Results
5.3.1.Gross effets of long term cadmium exposure
5.3.2.Trend of cadmium accumulation in plant and related seed ultrasctructural changes
5.3.3.Analysis of plant reproductive fitness
5.3.4.Physio-biochemical changes in silique wall
5.4.Discussion
5.5.Conclusion
CHAPTER6.Major findings and future perspectives
6.1.Major findings
6.2.Future perspectives
References
List of publications during PhD study period
Supplemental materials
本文编号：3893330
【文章页数】：159 页
【学位级别】：博士
【文章目录】：
Acknowledgements
List of abbreviations
Abstract
摘要
CHAPTER1.General Introduction and Literature Review
1.1.Cadmium occurrence in the environment
1.1.1.Worldwide occurrence of cadmium
1.1.2.Cadmium-soil contamination in China
1.2.Environmental issues of cadmium exposure
1.2.1.Cadmium phytotoxicity effects
1.2.2 Cadmium and human health risks
1.3.Brassica napus,a tool for sustainable management of Cd-contaminated soils
1.4.Current state of knowledge on Cd adaptive responses in Brassica napus
1.5.Research objectives
1.6.Outline of the study
CHAPTER2.Intraspecific variability of cadmium accumulation,subcellular distribution and chemical forms in Brassica napus
2.1.Introduction
2.2.Materials and methods
2.2.1.Plant material and experimental conditions
2.2.2.Determination of cadmium content
2.2.3.Plant tissues fractionation and metal analysis
2.2.4.Analysis of metal chemical forms
2.2.5.Statistical analysis
2.3.Results
2.3.1.Variation in cadmium-induced changes in plant biomass,and trend of metal accumulation
2.3.2.Metal subcellular distribution
2.3.3.Metal chemical forms
2.4.Discussion
2.5.Conclusion
CHAPTER3.Cadmium detoxification in high Cd-accumulating Brassica napus genotypes involves changes in cell wall compositional profiles
3.1.Introduction
3.2.Materials and methods
3.2.1.Plant material and experimental conditions
3.2.2.Cell wall extraction and polysaccharides fractionation
3.2.3.Measurement of uronic acid and total sugar in fractionated cell wall polysaccharides
3.2.4.Quantification of different fractions of pectin and estimation of its methyl-esterification degree
3.2.5.Determination of Cd contents in fractionated cell wall polysaccharides
3.2.6.Measurement of ascorbic acid content
3.2.7.Processing of cell wall-related RNA-seq data
3.2.7.1.Total RNA extraction,reliability assessment and RNA-sequence analyses
3.2.7.2.Verification of RNA-Seq data by qRT-PCR assay
3.2.8.Statistical analysis
3.3.Results
3.3.1.Cd-induced remodelling of cell wall polysaccharides among B.napus genotypes
3.3.2.Relative contribution of different cell wall sub-fractions to Cd immobilization
3.3.3.Cadmium influence on transcriptional regulation of pectin and hemicellulose biosynthesis
3.3.4.Pectin as possible substrat for ascorbic acid production in leaves
3.4.Discussion
3.5.Conclusion
CHAPTER4.Comparative metabolomic responses of low-and high-cadmium accumulating genotypes reveal the Cd adaptive mechanism in Brassica napus
4.1.Introduction
4.2.Materials and methods
4.2.1.Plant material and experimental conditions
4.2.2.Metabolomic analysis
4.2.2.1.Metabolites extraction and liquid chromatography-mass spectrometry
4.2.2.2.Data processing
4.2.3.Analysis of other physio-biochemical endpoints
4.2.3.1.Assessment of Cd accumulation in plant tissues
4.2.3.2.Transmission electron microscopy,scanning electron microscopy and chlorophyll fluorescence imaging
4.2.3.3.Determination of phenolic contents and total antioxidant capacity
4.2.3.4.Total RNA extraction,cDNA synthesis,and qRT-PCR assay
4.2.4.Statistical analyses
4.3.Results
4.3.1.Genotypic variation of responses to cadmium stress
4.3.2.General metabolomic changes in the two genotypes following Cd stress
4.3.3.Profiling of metabolites specifically regulated in either genotype under Cd stress
4.3.3.1.Metabolomic signature in plant primary metabolism
4.3.3.2.Metabolomic signature in plant secondary metabolism
4.4.Discussion
4.5.Conclusion
CHAPTER5.Cadmium-induced reproductive repercussions in two Brassica napus genotypes supplemented with serotonin,and glimpses into potential adaptive mechanisms
5.1.Introduction
5.2.Materials and methods
5.2.1.Plant material and experimental conditions
5.2.2.Data collection and samples preparation for analysis
5.2.3.Elements analysis in different plant parts
5.2.4.Seed oil content determination
5.2.5.Seed transmission electron microscopy
5.2.6.Evaluation of pollen grain viability and cross-pollination experiments
5.2.7.Measurement of other physio-biochemical end points
5.2.7.1.Determination of indices of oxidative stress
5.2.7.2.Photosynthetic parameters
5.2.7.3.Determination of antioxidant enzymes
5.2.7.4.Thiol compounds assays
5.2.7.5.Assays of activities of sulfur assimilation enzymes
5.2.7.6.Measurement of phenolic contents
5.2.8.Data analysis
5.3.Results
5.3.1.Gross effets of long term cadmium exposure
5.3.2.Trend of cadmium accumulation in plant and related seed ultrasctructural changes
5.3.3.Analysis of plant reproductive fitness
5.3.4.Physio-biochemical changes in silique wall
5.4.Discussion
5.5.Conclusion
CHAPTER6.Major findings and future perspectives
6.1.Major findings
6.2.Future perspectives
References
List of publications during PhD study period
Supplemental materials
本文编号：3893330
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