Proteomic analysis of Arabidopsis seed germination and priming
Proteomic analysis of Arabidopsis seed germination and priming
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DOI：
10.1104/pp.126.2.835
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发表时间：
2001-06-01
期刊：
影响因子：
7.4
通讯作者：
Job, D
中科院分区：
文献类型：
--
作者：
Gallardo, K;Job, C;Job, D
为了更好地了解种子萌发，一个复杂的发育过程，我们开发了一个蛋白质组分析的模式植物拟南芥的完整基因组序列，现在是可用的。在约1,300个总的种子蛋白在二维凝胶中解决，74个蛋白质的丰度（上调和下调）的变化，观察到严格意义上的萌发（即胚根出现之前）和胚根突起的步骤。这种方法也被用来分析蛋白质的变化发生在工业种子预处理，如引发，加速种子发芽，提高幼苗的均匀性。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。他们中的一些人以前已被证明在几种植物的发芽和/或引发过程中发挥作用，这一发现强调了使用拟南芥作为种子质量的分子分析的模型系统的有用性。此外，在蛋白质水平上进行的本研究验证了先前在基因表达水平上获得的结果（例如，从差异表达的mRNA的定量或启动子/报告子构建体的分析）。最后，这种方法揭示了与种子萌发和引发的不同阶段相关的新蛋白质。其中一些参与种子的吸胀过程（如肌动蛋白同种型或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）。这些事实突出了蛋白质组学的力量，以解开复杂的发育过程，如发芽的具体特点，并检测蛋白质标记，可用于表征种子活力的商业种子批次和开发和监测引发处理。
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.