Gallardo K, Job C, Groot S P, Puype M, Demol H, Vandekerckhove J, Job D
Laboratoire Mixte Centre National de la Recherche Scientifique-Institut National de la Recherche Agronomique-Aventis, Aventis CropScience, Lyon, France.
Plant Physiol. 2001 Jun;126(2):835-48. doi: 10.1104/pp.126.2.835.
To better understand seed germination, a complex developmental process, we developed a proteome analysis of the model plant Arabidopsis for which complete genome sequence is now available. Among about 1,300 total seed proteins resolved in two-dimensional gels, changes in the abundance (up- and down-regulation) of 74 proteins were observed during germination sensu stricto (i.e. prior to radicle emergence) and the radicle protrusion step. This approach was also used to analyze protein changes occurring during industrial seed pretreatments such as priming that accelerate seed germination and improve seedling uniformity. Several proteins were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry. Some of them had previously been shown to play a role during germination and/or priming in several plant species, a finding that underlines the usefulness of using Arabidopsis as a model system for molecular analysis of seed quality. Furthermore, the present study, carried out at the protein level, validates previous results obtained at the level of gene expression (e.g. from quantitation of differentially expressed mRNAs or analyses of promoter/reporter constructs). Finally, this approach revealed new proteins associated with the different phases of seed germination and priming. Some of them are involved either in the imbibition process of the seeds (such as an actin isoform or a WD-40 repeat protein) or in the seed dehydration process (e.g. cytosolic glyceraldehyde-3-phosphate dehydrogenase). These facts highlight the power of proteomics to unravel specific features of complex developmental processes such as germination and to detect protein markers that can be used to characterize seed vigor of commercial seed lots and to develop and monitor priming treatments.
为了更好地理解种子萌发这一复杂的发育过程,我们对模式植物拟南芥开展了蛋白质组分析,其完整基因组序列现已可得。在二维凝胶中分辨出的约1300种种子总蛋白中,在严格意义上的萌发期间(即胚根出现之前)和胚根突出阶段,观察到74种蛋白质丰度的变化(上调和下调)。该方法还用于分析工业种子预处理(如引发处理)过程中发生的蛋白质变化,引发处理可加速种子萌发并提高幼苗一致性。通过基质辅助激光解吸电离飞行时间质谱鉴定了几种蛋白质。其中一些蛋白质先前已被证明在几种植物物种的萌发和/或引发过程中发挥作用,这一发现突出了使用拟南芥作为种子质量分子分析模型系统的实用性。此外,本研究在蛋白质水平上进行,验证了先前在基因表达水平上获得的结果(例如,通过差异表达mRNA的定量或启动子/报告基因构建体的分析)。最后,这种方法揭示了与种子萌发和引发不同阶段相关的新蛋白质。其中一些蛋白质参与种子的吸水过程(如一种肌动蛋白异构体或一种WD-40重复蛋白)或种子脱水过程(如胞质甘油醛-3-磷酸脱氢酶)。这些事实凸显了蛋白质组学在揭示复杂发育过程(如萌发)的特定特征以及检测可用于表征商业种子批次种子活力和开发及监测引发处理的蛋白质标记方面的能力。