ABA（脫落酸）是一種重要的逆境激素,調節(jié)植株生長和發(fā)育的多個形態(tài)和生理方面。據報道,來自擬南芥的許多基因都參與了ABA信號轉導和脅迫調控,但是,一些ABA響應基因的功能仍然未知。在這項研究中,我們報道了擬南芥中作為ABA響應基因的AWR（ABA響應WD40）。qRT-PCR結果顯示,ABA處理,AWR的表達水平顯著增加。原生質體轉染試驗表明,AWR位于細胞質和細胞膜中。RT-PCR分析表明,AWR在莖中高表達,但在其他器官如根,蓮座葉和莖葉中也有表達。萌發(fā)實驗分析表明,AWR過表達植株對ABA更為敏感。在任何給定的濃度下,過表達植株的發(fā)芽速度分別比野生型和突變型發(fā)芽快。綠苗試驗表明,與Co 1野生型和突變型植株相比,35S:AWR轉基因植株對ABA更為敏感。NaCl分析表明,與野生型和突變型植株相比,AWR過表達植株對鹽脅迫更為敏感。形態(tài)表型觀察表明,與Col野生型和突變型植株相比,AWR過表達轉基因植株具有更長的根。根伸長實驗還表明,ABA處理后,過表達植株與其他株系相比的根更長。也報道,過表達植株在根伸長實驗中對NaCl處理更敏感。響應甘露醇實驗,過表達植株表現(xiàn)出比Col野生型和... 

【文章來源】：東北師范大學吉林省 211工程院校 教育部直屬院校

【文章頁數(shù)】：60 頁

【學位級別】：碩士

【文章目錄】：
Abstract
摘要
Introduction
    1.Abscisic Acid（ABA）
        1.1.ABA signalling in plants
        1.2.ABA Binding Proteins and Receptors
        1.3.Role of ABA in stress signaling
    2.ABA and abiotic stress tolerance
        2.1.ABA and salt tolerance
        2.2.ABA and drought tolerance
        2.3.Effect of ABA on seed germination and root elongation
    3.WD40 proteins and ABA signalling
    4.Purpose and significance of this study
Materials and methods
    1.Experimental Materials
        1.1.Plants Materials
        1.2.Bacterium strain and plasmids
        1.3.List of Conventional reagents used
        1.4.Restriction enzymes
        1.5.Preparation of Growth Medium
        1.6.Preparation of Antibiotics and buffers
    2.Designing primers
        2.1.Selection of target sites
        2.2.Protocol
    3.Equipments
    4.EXPERIMANTAL METHODS
        4.1.Sowing seed and vernalization
        4.2.Bioinformatics analysis
        4.3.Plant RNA extraction
        4.4.Vector construction and cloning
        4.5.Agrobacterium tumefeciens competent cell transformation
        4.6.Plants transformation,selection of transgenic plants and mutant isolation
        4.7.Plant DNA isolation/extraction
        4.8.Large-scale extraction of plasmid DNA
        4.9.Protoplast transfection assay
        4.10.Protein localization observation
        4.11.Treatment of hormone
        4.12.Salinity stress
        4.13.Drought stress
Results
    1.ARW is ABA responsive gene
    2.Cloning of ARW
    3.Construction of p PZP-HA-ARW Plasmid
    4.Subcellular localization of ARW
        4.1.Construction of p UC19-GFP-ARW
        4.2.ARW is located in the cell membrane and cytosol
    5.Expression pattern of ARW
    6.ARW overexpression plants
    7.ARW mutant plants
    8.Hormone treatment
        8.1.Effect of ABA on seed germination rate
        8.2.Effect of ABA on Cotyledon Greening rate
        8.3.Effect of ABA on root elongation
    9.Effect of Na Cl on root elongation
    10.Effect of Mannitol on root elongation
    11.Rosette leaves size
    12.Leaf numbers and area
Discussion
Conclusion
References



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