葡萄是一種廣泛栽植的重要果樹,其生長(zhǎng)發(fā)育易受世界范圍內(nèi)非生物和生物脅迫的影響。干旱脅迫和銅脅迫是兩種重要的非生物脅迫,也是葡萄栽培生產(chǎn)中的主要限制因素。因此,有必要去挖掘能夠抵御或者適應(yīng)這些逆境的遺傳資源。干旱脅迫下葡萄轉(zhuǎn)錄水平的測(cè)定:對(duì)旱干脅迫處理組和對(duì)照組構(gòu)建的cDNA文庫進(jìn)行RNA測(cè)序（RNA-seq）分析,結(jié)果顯示,在發(fā)現(xiàn)的12451個(gè)差異表達(dá)的基因（DEGs）中,有8021個(gè)基因上調(diào)表達(dá),4430個(gè)基因表達(dá)下調(diào)。干旱脅迫下生理指標(biāo)的測(cè)定:葉片氣孔導(dǎo)度的下降抑制了光合活性和CO2同化作用,這與轉(zhuǎn)錄組分析結(jié)果一致�；钚匝跸到y(tǒng)（包括應(yīng)激酶及其相關(guān)蛋白）和次生代謝反應(yīng)被激活,以適應(yīng)干旱脅迫。同時(shí),葡萄中脫落酸（ABA）、赤霉素（GA）、生長(zhǎng)素（IAA）、油菜素內(nèi)酯（BR）等激素也會(huì)參與到葡萄應(yīng)激干旱脅迫的過程中。干旱條件下脅迫應(yīng)答信號(hào)網(wǎng)絡(luò)的構(gòu)建:干旱脅迫抑制了光合作用過程,影響了干旱應(yīng)答脅迫葡萄葉片中還原糖（葡萄糖和果糖）的合成。葡萄葉片內(nèi)氧化應(yīng)激標(biāo)記物（丙二醛,MDA）引起了超氧自由基（02-）和過氧化氫（H2O2）的升高,植物體抗氧化清除系統(tǒng)激活以對(duì)抗活性氧（ROS）。鈣離子（C... 

【文章來源】：南京農(nóng)業(yè)大學(xué)江蘇省 211工程院校 教育部直屬院校

【文章頁數(shù)】：154 頁

【學(xué)位級(jí)別】：博士

【文章目錄】：
ABSTRACT
摘要
LIST OF ABBREVIATIONS
CHAPTER 1 Introduction and Review of Literature
    1. Background
    2. Overview of drought and Cu stress: plant responses and mechanism of acclimation
    3. Physiological responses
        3.1 Osmotic adjustment
        3.2 Stomatal signaling
        3.3 Reactive oxygen species (ROS) signaling
        3.4 Plant hormone signaling
        3.5 Mitogen-activated protein kinase (MAPK) signaling
        3.6 Role of transcription factors (TFs) in abiotic stress tolerance
        3.7 Role of autophagy in abiotic stress tolerance
    4 Implication of Omics approaches in abiotic stress
        4.1 Transcriptomics
        4.2 Metabolomics
    5.Grapevine in a changing environment
        5.1 Drought stress
        5.2 Cu Stress
    6.Objectives of the study
    References
CHAPTER 2 Insights into grapevine defense response against drought as revealed bybiochemical, physiological and RNA-seq analysis
    1. Introduction
    2 Materials and Methods
        2.1 Plant material and drought treatments
        2.2 Determination of important biochemistry and physiology-related traits
        2.3 RNA extraction, cDNA library construction and Illumina deep sequencing
        2.4 Analysis of gene expression level, gene ontology (GO) and Kyotoencyclopedia of genes and genomics (KEGG)
        2.5 Illumina RNA-seq results validation by qRT-PCR
    3. Results
        3.1. Gene ontology (GO) and KEGG analysis of differentially expressed genes
        3.2. Chlorophyll degradation and photosynthetic competencies under droughtstress
        3.3 ROS system under drought stress
        3.4 Plant hormone signal transduction pathway under drought stress
        3.5 Proline metabolism under drought stress
        3.6 Biosynthesis of secondary metabolites under drought stress
        3.7 Heat shock protein (HSP) and pathogenesis-related protein (PR) in response todrought stress
        3.8 qRT-PCR validation of DEGs from Illumina RNA-Seq
    4 Discussion
    5. Conclusion
    References
CHAPTER 3 Grapevine immune signaling network in response to drought stress asrevealed by transcriptomic analysis
    1. Introduction
    2. Materials and Methods
        2.1 Plant material and drought treatment
        2.2 RNA extraction,cDNA library construction and Illumina deep sequencing
        2.3 Analysis of gene expression level, Gene Ontology (GO) and KyotoEncyclopedia of Gene and Genome (KEGG)
        2.4 Estimation of important physiological and biochemical parameters
        2.5 Illumina RNA-Seq results validation by qRT-PCR
    3. Results
        3.1 GO and KEGG analysis of differentially-expressed genes in response todrought stress
        3.2 Effect of drought stress on physiological and biochemical parameters ofgrapevine
        3.3 Calcium signaling pathway in response to drought stress
        3.4. MAPK-signaling pathway in response to drought stress
        3.5. Plant-pathogen interaction pathway in response to drought stress
        3.6. qRT-PCR validation of differentially expressed genes from Illumina RNA-Seq
    4. Discussion
    5. Conclusion
    References
CHAPTER 4 Genome-wide analysis of autophagy-related genes (ARGs) in grapevine andplant tolerance to copper stress
    1. Introduction
    2. Material and methods
        2.1. Plant material
        2.2 Identification of autophagy-related genes (ARGs) in grapevine
        2.3 Bioinformatics analysis of autophagy-related genes (ARGs)
        2.4 VvARGs expression profiles
        2.5 Autophagosome monitoring and malondialdehyde measuring in grapevineleaves
        2.6 RNA isolation and RT-qPCR
    3. Results
        3.1 Identification of 35 autophagy-related genes (ARGs) in the grape genome
        3.2 family conservation and expansion of VvARG homologues
        3.3 Duplication events and divergence rates of VvARG genes
        3.4 Grape ARG expression profiles at various fruit developmental stages
        3.5 Grape ARG gene expression profiles under abiotic stresses
        3.6 Autophagy monitoring and MDA measuring in the grape leaf under copperstress
        3.7 RT-qPCR of grapevine ARG genes
    4 Discussion
        4.1 Bioinformatics analysis of VvARG genes
        4.2 Expression profile of grapevine ARGs
        4.3 Copper and autophagy
    5 Conclusion
    References
RESEARCH OUT COMES AND FUTURE PERSPECTIVE
PUBLICATION
ACKNOWLEDGEMENTS


【參考文獻(xiàn)】：
期刊論文
[1]適于葡萄不同組織RNA提取方法的篩選[J]. 張彥蘋,王晨,于華平,蔡斌華,房經(jīng)貴.  西北農(nóng)業(yè)學(xué)報(bào). 2010(11)
[2]Arabidopsis AtBECLIN 1/AtAtg6/AtVps30 is essential for pollen germination and plant development[J]. Genji Qin~1 Zhiqiang Ma~1 Li Zhang~1 Shufan Xing~1 Xianhui Hou~1 Jie Deng~1 Jingjing Liu~1 Zhangliang Chen~(1,2) Li-Jia Qu~(1,2) Hongya Gu~(1,2) ~1National Laboratory for Protein Engineering and Plant Genetic Engineering,Peking- Yale Joint Research Center for Plant Molecular Genetics and AgroBiotechnology,College of Life Sciences,Peking University,Beijing 100871,China; ~2The National Plant Gene Research Center (Beijing),Beijing 100101,China.  Cell Research. 2007(03)



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