Flavonoids Enhancement and Alleviation of Stress in Brassica
发布时间：2022-07-27 20:35
小白菜（Brassica campestris ssp.chinensis L.）,属于十字花科,含较高的纤维素等植物化学物质,具有显著的促进健康的特性,是中国最主要的绿叶蔬菜。新鲜蔬菜的质量可以根据其营养价值、生长条件和肥料的施用情况进行评估。次生代谢物是植物中主要的植物化学物质,对植物的生存和人类健康都起着重要的作用。类黄酮,是一类广泛分布的多酚次生代谢物,具有广泛的生物活性,与植物生理、生长和发育相关。类黄酮,具有很强的抗氧化活性,能保护植物免受不利因子的胁迫,还能促进与植物和微生物的相互作用。类黄酮最重要的作用之一是保护植物免受非生物（盐、干旱、紫外辐射和高温）和生物（病原体和害虫）胁迫。类黄酮分子中共轭双键的存在使它们具有显著的抗氧化活性,而清除活性氧对于维持植物细胞内氧化还原状态的平衡非常重要。一般情况下,黄酮类化合物能够通过还原单线态氧,阻碍活性氧合成酶（如脂氧合酶、环氧合酶黄嘌呤氧化酶、单氧合酶）活性,螯合金属离子,抑制脂质过氧化等方式抑制活性氧的生成,并促进其他抗氧化剂的循环。在植物中,类黄酮的大部分功能是由于其强大的抗氧化特性。类黄酮不仅参与植物对抗不利环境条件,有...
【文章页数】：162 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
Chapter 1 General introduction and literature review
1.1 Beneficial phytochemicals in Pakchoi
1.2 Potential roles of flavonoids in Rhizosphere
1.3 Flavonoids and symbionts recruitment
1.4 Flavonoids and environmental conditions
1.5 Piriformospora indica
1.6 Host plant root colonization by P.indica
1.7 Biotic and abiotic stress tolerance
1.8 Biocontrol effect of Periformospora indica
1.9 P.indica alter the expression level of defense and stress-related genes
1.10 Concluding remarks about beneficial fungus P.indica
1.11 Study goal
1.12 Specific objectives
1.13 Research framework
Chapter 2 Elevation of secondary metabolites synthesis in Brassica campestris ssp.chinensis L.via exogenous inoculation of Piriformospora indica with appropriate fertilizer
2.1 Introduction
2.2 Materials and methods
2.2.1 Chemicals and reagents
2.2.2 Greenhouse experiments
2.2.3 Fertilizers and inoculation of soil
2.2.4 Fungal inoculum preparation
2.2.5 Plant material and growth conditions
2.2.6 Total RNA extraction and cDNA synthesis
2.2.7 Quantitative real-time PCR analysis
2.2.8 Growth and root analysis
2.2.9 Measurement of total phenolic compounds
2.2.10 Measurement of total flavonoid content
2.2.11 Measurement of phenolic acid content
2.2.12 HPLC-MS analysis
2.2.13 Chlorophylls and carotenoids measurement
2.2.14 Total antioxidant activity
2.2.15 Statistical analysis of data
2.3 Results
2.3.1 Morphological indices
2.3.2 Roots colonization assay
2.3.3 Health-promoting compounds analysis
2.3.4 Chlorophylls and carotenoids
2.3.5 Expression level of flavonoid pathway genes
2.3.6 Free radical scavenging activity
2.4 Discussion
2.5 Conclusions
2.6 Abbreviations used
Chapter 3 An endosymbiont Piriformospora indica reduces adverse effects of salinity by regulating cation transporter genes,phytohormones,and antioxidants in Brassica campestris ssp.chinensis
3.1 Introduction
3.2 Material and methods
3.2.1 Piriformospora indica culture and plant growth
3.2.2 Treatments
3.2.3 Parameters measured
3.2.3.1 Biomass production
3.2.3.2 Root colonization assay
3.2.3.3 Total chlorophyll assessment
3.2.3.4 Leaf relative water content and electrolyte leakage
3.2.3.5 Efficiency of photosystem II
3.2.3.6 Antioxidant enzyme assay and malondialdehyde(MDA)estimation
3.2.3.7 Determination of Phytohormones by UPLC-3QMS
3.2.3.8 Mineral nutrients determination
3.2.3.9 Total RNA extraction and cDNA synthesis
3.2.3.10 Quantitative real-time PCR analysis
3.2.4 Statistical analysis of data
3.3 Results
3.3.1 P.indica promoted B.campestris ssp.chinensis growth
3.3.2 P.indica root colonization/symbiotic development assay
3.3.3 P.indica increased leaf area and chlorophyll content
3.3.4 Electrolyte leakage,relative water content,and efficiency of photosystem II
3.3.5 P.indica improved the enzymatic activity under saline conditions
3.3.6 P.indica reduced leaf MDA content under saline conditions
3.3.7 P.indica augmented abscisic acid(ABA)and salicylic acid(SA)concentration
3.3.8 P.indica enhances the uptake of phosphorus and reduce sodium accumulation under saline conditions
3.3.9 Encoded ion transporter genes expression analysis
3.4 Discussion
Chapter 4 Suppression of clubroot(Plasmodiophora brassicae) development in Brassica campestris ssp. chinensis L. via exogenous inoculation of Piriformospora indica
4.1 Introduction
4.2 Material and methods
4.2.1 Piriformospora indica and Plasmodiophora brassicae
4.2.2 Plant cultivation and inoculation
4.2.3 Properties of Soil and growth conditions
4.2.4 Morphological and phytopathological analysis
4.2.5 Major flavonoids determination
4.2.6 Molecular analysis
4.2.7 Statistical analysis
4.3 Results
4.3.1 Morphological assessment
4.3.2 Root colonization
4.3.3 Polyphenols
4.3.4 Clubroot symptoms under the influence of P.indica
4.3.5 P.brassicae genes expression analysis through RT.PCR
4.4 Discussion
4.5 Conclusion
Chapter 5 Transcriptomic response of Pakchoi after incubation with endosymbiont Periformosopora indica,a possible biocontrol agent of Plasmodiophora brassicae
5.1 Introduction
5.2 Materials and methods
5.2.1 Piriformospora indica and Plasmodiophora brassicae
5.2.2 Plant cultivation and inoculation
5.2.3 Properties of Soil and growth conditions
5.2.4 RNA Isolation
5.2.5 Differential expression and mapping:Sequencing results analysis
5.2.6 Functional analysis of differentially expressed genes
5.2.7 Validation of RNA-Seq data by quantitative real-time PCR
5.3 Results
5.3.1 Roots sampling and confirmation of successful colonization by P.indica
5.3.2 RNA-Seq Analysis
5.3.3 NR annotation statistics
5.3.4 Analysis and Verification of Differentially Expressed Genes(DEGs)
5.3.5 Functional annotation analysis of DEGs
5.3.6 Pathways analysis
5.3.7 KEGG pathway enrichment analysis in each treatment
5.4 Discussion
Chapter 6 Major findings and outlook
6.1 Major findings
6.2 Outlook
References
List of publications
Funding information
Acknowledgement
本文编号：3666131
【文章页数】：162 页
【学位级别】：博士
【文章目录】：
Abstract
摘要
Chapter 1 General introduction and literature review
1.1 Beneficial phytochemicals in Pakchoi
1.2 Potential roles of flavonoids in Rhizosphere
1.3 Flavonoids and symbionts recruitment
1.4 Flavonoids and environmental conditions
1.5 Piriformospora indica
1.6 Host plant root colonization by P.indica
1.7 Biotic and abiotic stress tolerance
1.8 Biocontrol effect of Periformospora indica
1.9 P.indica alter the expression level of defense and stress-related genes
1.10 Concluding remarks about beneficial fungus P.indica
1.11 Study goal
1.12 Specific objectives
1.13 Research framework
Chapter 2 Elevation of secondary metabolites synthesis in Brassica campestris ssp.chinensis L.via exogenous inoculation of Piriformospora indica with appropriate fertilizer
2.1 Introduction
2.2 Materials and methods
2.2.1 Chemicals and reagents
2.2.2 Greenhouse experiments
2.2.3 Fertilizers and inoculation of soil
2.2.4 Fungal inoculum preparation
2.2.5 Plant material and growth conditions
2.2.6 Total RNA extraction and cDNA synthesis
2.2.7 Quantitative real-time PCR analysis
2.2.8 Growth and root analysis
2.2.9 Measurement of total phenolic compounds
2.2.10 Measurement of total flavonoid content
2.2.11 Measurement of phenolic acid content
2.2.12 HPLC-MS analysis
2.2.13 Chlorophylls and carotenoids measurement
2.2.14 Total antioxidant activity
2.2.15 Statistical analysis of data
2.3 Results
2.3.1 Morphological indices
2.3.2 Roots colonization assay
2.3.3 Health-promoting compounds analysis
2.3.4 Chlorophylls and carotenoids
2.3.5 Expression level of flavonoid pathway genes
2.3.6 Free radical scavenging activity
2.4 Discussion
2.5 Conclusions
2.6 Abbreviations used
Chapter 3 An endosymbiont Piriformospora indica reduces adverse effects of salinity by regulating cation transporter genes,phytohormones,and antioxidants in Brassica campestris ssp.chinensis
3.1 Introduction
3.2 Material and methods
3.2.1 Piriformospora indica culture and plant growth
3.2.2 Treatments
3.2.3 Parameters measured
3.2.3.1 Biomass production
3.2.3.2 Root colonization assay
3.2.3.3 Total chlorophyll assessment
3.2.3.4 Leaf relative water content and electrolyte leakage
3.2.3.5 Efficiency of photosystem II
3.2.3.6 Antioxidant enzyme assay and malondialdehyde(MDA)estimation
3.2.3.7 Determination of Phytohormones by UPLC-3QMS
3.2.3.8 Mineral nutrients determination
3.2.3.9 Total RNA extraction and cDNA synthesis
3.2.3.10 Quantitative real-time PCR analysis
3.2.4 Statistical analysis of data
3.3 Results
3.3.1 P.indica promoted B.campestris ssp.chinensis growth
3.3.2 P.indica root colonization/symbiotic development assay
3.3.3 P.indica increased leaf area and chlorophyll content
3.3.4 Electrolyte leakage,relative water content,and efficiency of photosystem II
3.3.5 P.indica improved the enzymatic activity under saline conditions
3.3.6 P.indica reduced leaf MDA content under saline conditions
3.3.7 P.indica augmented abscisic acid(ABA)and salicylic acid(SA)concentration
3.3.8 P.indica enhances the uptake of phosphorus and reduce sodium accumulation under saline conditions
3.3.9 Encoded ion transporter genes expression analysis
3.4 Discussion
Chapter 4 Suppression of clubroot(Plasmodiophora brassicae) development in Brassica campestris ssp. chinensis L. via exogenous inoculation of Piriformospora indica
4.1 Introduction
4.2 Material and methods
4.2.1 Piriformospora indica and Plasmodiophora brassicae
4.2.2 Plant cultivation and inoculation
4.2.3 Properties of Soil and growth conditions
4.2.4 Morphological and phytopathological analysis
4.2.5 Major flavonoids determination
4.2.6 Molecular analysis
4.2.7 Statistical analysis
4.3 Results
4.3.1 Morphological assessment
4.3.2 Root colonization
4.3.3 Polyphenols
4.3.4 Clubroot symptoms under the influence of P.indica
4.3.5 P.brassicae genes expression analysis through RT.PCR
4.4 Discussion
4.5 Conclusion
Chapter 5 Transcriptomic response of Pakchoi after incubation with endosymbiont Periformosopora indica,a possible biocontrol agent of Plasmodiophora brassicae
5.1 Introduction
5.2 Materials and methods
5.2.1 Piriformospora indica and Plasmodiophora brassicae
5.2.2 Plant cultivation and inoculation
5.2.3 Properties of Soil and growth conditions
5.2.4 RNA Isolation
5.2.5 Differential expression and mapping:Sequencing results analysis
5.2.6 Functional analysis of differentially expressed genes
5.2.7 Validation of RNA-Seq data by quantitative real-time PCR
5.3 Results
5.3.1 Roots sampling and confirmation of successful colonization by P.indica
5.3.2 RNA-Seq Analysis
5.3.3 NR annotation statistics
5.3.4 Analysis and Verification of Differentially Expressed Genes(DEGs)
5.3.5 Functional annotation analysis of DEGs
5.3.6 Pathways analysis
5.3.7 KEGG pathway enrichment analysis in each treatment
5.4 Discussion
Chapter 6 Major findings and outlook
6.1 Major findings
6.2 Outlook
References
List of publications
Funding information
Acknowledgement
本文编号：3666131
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