葡萄是世界范圍內(nèi)廣泛栽種的水果作物之一。中國作為葡萄的起源中心之一,擁有極其豐富的種質(zhì)資源。溫度是決定葡萄生長發(fā)育和地理分布的主要因素之一。在中國北方釀酒葡萄主產(chǎn)區(qū),葡萄會遭受冬季極端低溫的影響,需要埋土防寒以度過嚴(yán)冬。高抗寒性已成為這些地區(qū)葡萄品種篩選的主要目標(biāo)。冷馴化是植物面對低溫脅迫的一種適應(yīng)策略,包括保護(hù)性蛋白質(zhì)的積累等多個過程。與主栽的歐亞種葡萄(V.vinifera)相比,原產(chǎn)我國的山葡萄(V.amurensis)具有極強(qiáng)的耐寒性。解析山葡萄耐寒機(jī)制將為葡萄的抗性育種提供理論基礎(chǔ)。本研究通過對山葡萄‘左山一號’和歐亞種‘京早晶’兩種葡萄在冷馴化過程中的芽組織進(jìn)行蛋白質(zhì)組分析,以探討兩個葡萄品種在田間冷馴化的耐寒機(jī)制,并進(jìn)一步選擇低溫應(yīng)答蛋白Snakin-2進(jìn)行功能分析。主要研究結(jié)果如下:1.葡萄休眠芽樣品于2016年10月19號和12月2號在中國科學(xué)院植物研究所采集。利用差熱分析發(fā)現(xiàn),葡萄休眠芽的抗寒性隨田間溫度的逐步降低而增加,‘左山一號’的抗寒性明顯高于‘京早晶’。因此擬通過樣品之間的比較來揭示冷馴化下兩個品種休眠芽在蛋白質(zhì)組學(xué)水平的差異。2.通過比較蛋白質(zhì)組學(xué)的方法,...

【文章頁數(shù)】：113 頁

【學(xué)位級別】：博士

【文章目錄】：
摘要
abstract
List of Abbreviations
Chapter 1 Introduction and Literature Review
    1.1 Background
    1.2 Grapevine breeding history in China
    1.3 Acclimation to low temperature
    1.4 Plant Proteomics
        1.4.1 Overview of iTRAQ-based Proteomics Workflow
        1.4.2 Mass Spectrometry
        1.4.3 Proteomics studies in grapevine
    1.5 Candidate protein for cold acclimation
        1.5.1 Chemical structure of Snakin/GASA protein
    1.6 Research Aims
Chapter 2 iTRAQ-based proteomics uncovers the variation in budsof Vitis amurensis and Vitis vinifera during cold acclimation
    2.1 Introduction
    2.2 Materials and Methods
        2.2.1 Plant materials
        2.2.2 Protein extraction,in-solution digestion,and peptide extraction
        2.2.3 iTRAQ labeling and high-performance liquid chromatography(HPLC)fractionation
        2.2.4 Nano-LC-MS/MS analysis
        2.2.5 MS/MS data analysis and quality control
        2.2.6 Hierarchical clustering analysis and functional annotation of theproteins
        2.2.7 qRT-PCR analysis
        2.2.8 Statistical analysis
    2.3 Results
        2.3.1.Overview of the quantitative proteomics data
        2.3.2.Mass Spectrometry Quality Control
        2.3.3.PCA of DAPs during cold acclimation
        2.3.4.Analysis of the10 most upregulated DAPs in V.amurensis and V.vinifera during CA
        2.3.5.Functional categories of the overlapping CA-related DAPs in V.amurensis and V.vinifera
        2.3.6.Pathways specifically enriched in V.amurensis and V.vinifera duringCA
        2.3.7 The phenylpropanoid biosynthesis pathway is differently regulated inthe two grapevines during CA
        2.3.8 A qRT-PCR analysis revealed the correlation between transcriptionexpression and proteomic levels
    2.4 Discussion
        2.4.1 Carbohydrate and energy metabolism
        2.4.2 Stress response and defense-related proteins
        2.4.3 Protein biosynthesis and metabolism
        2.4.4 Phenylpropanoid biosynthesis pathway
    2.5 Correlation between transcriptional and proteomic expression
Chapter 3 Functional Analysis of the CA related protein Snakin-2
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Materials preparation
        3.2.2 RNA extraction and quantitative reverse transcription PCR(qRT-PCR)analysis
        3.2.3 Vector construction for gene transformation
        3.2.4 Transformation and generation transgenic callus from petiolesegments
        3.2.5 Molecular analysis of Vasnk-2 transgenic callus lines
        3.2.6 Subcellular localization of Snakin-2
    3.3 Results
        3.3.1 Quantitative expression of Snk/GASA2 after cold treatment
        3.3.2 Vector construction and gene transformation for overexpression in V.amurensis
        3.3.3.Molecular analysis of the transformed calli
    3.7 Discussion
Chapter 4 Conclusions and Future Prospects
    4.1 Conclusions
    4.2 Future prospects
References
Appendix
Acknowledgements
Student profile and publications



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