二穗短柄草屬于禾本科,它與主要糧食作物如包括小麥、大麥和黑麥有密切的系統(tǒng)進(jìn)化關(guān)系。二穗短柄草因其基因組小、生長(zhǎng)周期短等優(yōu)點(diǎn)而廣泛用于研究。本研究在二穗短柄草DREB基因家族分析的基礎(chǔ)上鑒定了58個(gè)BdDREB基因,根據(jù)進(jìn)化關(guān)系可分為A1、A2、A3、A4、A5和A6六個(gè)亞組。染色體定位分析發(fā)現(xiàn)它們主要定位在二穗短柄草五個(gè)染色體上。對(duì)不同組織和不同非生物脅迫條件下表達(dá)模式分析發(fā)現(xiàn)他們主要參與植物的生長(zhǎng)發(fā)育。此外,我們還成功克隆了BdDREB基因（XM₀03570049.3）,并成功在擬南芥中介導(dǎo)表達(dá)且篩選到一定數(shù)量的轉(zhuǎn)基因擬南芥種子,最后用PCR檢測(cè)和表型分析了轉(zhuǎn)基因擬南芥植物,為進(jìn)一步研究BdDERB的功能奠定了基礎(chǔ)。 

【文章來(lái)源】：西北農(nóng)林科技大學(xué)陜西省 211工程院校 985工程院校 教育部直屬院校

【文章頁(yè)數(shù)】：59 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
ABSTRACT
摘要
Chapter one: Review of Literature
    1.1 Introduction
    1.2 Transcription factors (TFs)
        1.2.1 AP2/EREBP transcription factors
        1.2.2 AREB transcription factors
        1.2.3 NAC transcription factors
        1.2.4 WRKY transcription factors
        1.2.5 Zinc finger transcription factors
    1.3 Arabidopsis thaliana: the model plant
    1.4 Abiotic stresses
        1.4.1 Drought stress
        1.4.2 Salinity stress
        1.4.3 Cold stress
        1.4.4 Heat stress
    1.5 Research objectives and outline of the experimental design
        1.5.1 Research objectives
        1.5.2 Flow chart of research process
Chapter Two: Materials and Methods
    2.1 Materials
        2.1.1 Plant Materials and Growth Conditions
            2.1.1.1 Brachypodium distachyon Growth Conditions
            2.1.1.2 Arabidopsis thaliana Growth Conditions
            2.1.1.3 Sterilization Arabidopsis seeds
        2.1.2 Database software and online tools
        2.1.3 List of primers used in the present study
    2.2 Methods
        2.2.1 Bd DREB gene analysis
            2.2.1.1 Multiple sequence alignment and phylogenetic analysis
            2.2.1.2 Chromosome distribution analysis
        2.2.2 Overexpression Bd DREB1G gene in Arabidopsis
            2.2.2.1 General sterilization procedures
            2.2.2.2 Recombinant DNA techniques for cloning and DNA analysis
                2.2.2.2.1 RNA isolation and c DNA synthesis
                2.2.2.2.2 Quantitative real-time PCR analysis
            2.2.2.3 Gene cloning
            2.2.2.4 PCR product purification
            2.2.2.5 T/A cloning of PCR products
            2.2.2.6 Preparation of ultra-competent bacterial cells
            2.2.2.7 Transformation to E.coli
            2.2.2.8 Plasmid extraction
            2.2.2.9 Restriction enzymes digestion and ligation
                2.2.2.9.1 Plasmid extraction
                2.2.2.9.2 Agrobacterium
                2.2.2.9.3 Transformation to arabidopsis with MS media
                2.2.2.9.4 Transgenic plants
                    2.2.2.9.4.1 Arabidopsis transformation
                2.2.2.9.5 Screening of transgenic Arabidopsis and DNA extraction
        2.2.3 Statistical analysis
Chapter Three: Results and discussion
    3.1 Results
        3.1.1 the analysis of Bd DREB gene family
            3.1.1.1 Phylogenetic analysis
            3.1.1.2 Gene duplication and syntey analysis of Bd AP2 genes
        3.1.2 overexpression of Bd DREB1G in Arabidopsis thaliana
            3.1.2.1 Pre- experimental
            3.1.2.2 Cloning and sequence analysis of the Brachypodium DREB gene
            3.1.2.3 Screening and identification of transgenic Arabidopsis thaliana
                3.1.2.3.1 Screening test resistance to kanamycin
                3.1.2.3.2 PCR identified transgenic seedlings Arabidopsis
            3.1.2.4 Detection of gene expression in transgenic Arabidopsis thaliana
            3.1.2.5 Phenotypic analysis of transgenic Arabidopsis plants
    3.2 Discussion
Chapter Four: conclusions and recommendations
References
Acknowledgements
CURRICULUM VITAE



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