ABA Regulates the Response of ARW in Abiotic Stress Toleranc

发布时间：2021-04-01 05:09

　　ABA（脱落酸）是一种重要的逆境激素,调节植株生长和发育的多个形态和生理方面。据报道,来自拟南芥的许多基因都参与了ABA信号转导和胁迫调控,但是,一些ABA响应基因的功能仍然未知。在这项研究中,我们报道了拟南芥中作为ABA响应基因的AWR（ABA响应WD40）。qRT-PCR结果显示,ABA处理,AWR的表达水平显着增加。原生质体转染试验表明,AWR位于细胞质和细胞膜中。RT-PCR分析表明,AWR在茎中高表达,但在其他器官如根,莲座叶和茎叶中也有表达。萌发实验分析表明,AWR过表达植株对ABA更为敏感。在任何给定的浓度下,过表达植株的发芽速度分别比野生型和突变型发芽快。绿苗试验表明,与Co 1野生型和突变型植株相比,35S:AWR转基因植株对ABA更为敏感。NaCl分析表明,与野生型和突变型植株相比,AWR过表达植株对盐胁迫更为敏感。形态表型观察表明,与Col野生型和突变型植株相比,AWR过表达转基因植株具有更长的根。根伸长实验还表明,ABA处理后,过表达植株与其他株系相比的根更长。也报道,过表达植株在根伸长实验中对NaCl处理更敏感。响应甘露醇实验,过表达植株表现出比Col野生型和... 

【文章来源】：东北师范大学吉林省 211工程院校 教育部直属院校

【文章页数】：60 页

【学位级别】：硕士

【文章目录】：
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



本文编号：3112728