干旱胁迫下萝卜种子萌发期蛋白的双向电泳分析

Two-Dimensional Electrophoresis Analysis of the Raphanus sativus L.Proteins During Seed Germination Under Drought Stress

种子萌发是植物生长、发育的最初阶段,也是容易遭到包括干旱在内的非生物胁迫的重要时期。蛋白组学是解析植物响应逆境胁迫的主要策略之一,双向电泳分离是蛋白组学的重要手段和环节。本研究以“春不老”萝卜为材料,利用聚乙二醇-6000(PEG-6000)模拟干旱胁迫,对干旱胁迫下萝卜种子萌发早期无子叶胚蛋白进行双向电泳分离和蛋白图谱差异分析。采取直接对萝卜无子叶胚体样品进行裂解和上样的方法,可得到分辨度和重复性较好的双向电泳蛋白图谱;对干旱胁迫下的萝卜种子萌发早期无子叶胚蛋白表达图谱进行差异分析,共检测和匹配265个萝卜蛋白点,响应干旱胁迫的蛋白质点有28个,其中13个显著上调表达,15个显著下调表达。研究结果表明,部分萝卜种子萌发期蛋白响应了干旱胁迫,为后续的蛋白质谱分析和响应蛋白的鉴定工作打下良好基础。本研究采用的对植物样品直接裂解和上样的方法,也为其他植物蛋白的双向电泳分析提供了借鉴。

Seed germination is the initial stage of plant growth and an important period of susceptibility to abiotic stress,including drought.Proteomics is one of the major tools for analyzing plant responses to stress,and twodimensional electrophoresis separation is an important means and step in proteomics.In the study,the radicle proteins of“Chunbulao”(Raphanus sativus L.)during germination period under drought stress simulating by Polyethylene Glycol-600(PEG-6000)are separated by two-dimensional electrophoresis.By directly lysising and loading radish non-cotyledons,a two-dimensional electrophoresis protein map with good resolution and repeatability is obtained;differential analysis is conducted on the expression profile of non-cotyledon proteins in radish during seed germination under drought stress,a total of 265 radish protein points are detected and matched,with 28 protein points responded to drought stress,of which 13 are significantly upregulated and 15 are significantly downregulated.The research results indicate that a few of radish seed germination proteins respond to drought stress and laid a solid foundation for subsequent protein mass spectrometry analysis and identification of responsive proteins.In addition,the method of direct lysis of samples used in this study also provides reference for two-dimensional electrophoresis analysis of other plant proteins.