Proteome analysis of grain filling and seed maturation in barley
Proteome analysis of grain filling and seed maturation in barley
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DOI：
10.1104/pp.003681
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发表时间：
2002-07-01
期刊：
影响因子：
7.4
通讯作者：
Svensson, B
中科院分区：
文献类型：
--
作者：
Finnie, C;Melchior, S;Svensson, B
在单子叶植物中，种子发育的过程包括淀粉胚乳中储备的沉积以及胚和糊粉层的发育。种子发育的最后阶段伴随着干燥耐受性的增加和成熟种子的干燥。我们已经使用了二维凝胶电泳的时间分辨的蛋白质的变化，发生在大麦种子发育过程中的研究（大麦）。大约1,000个低盐可提取的蛋白质点可以在二维凝胶上分辨。根据5周比较期间出现或消失的时间，将蛋白质斑点分为6类。19种不同的蛋白质或蛋白质片段在36个选定的点进行了鉴定基质辅助激光解吸电离飞行时间质谱（MS）或纳米电喷雾串联MS/MS。一些蛋白质存在于整个发展（例如，胞质苹果酸脱氢酶），而其他与早期籽粒灌浆（抗坏血酸过氧化物酶）或干燥（Cor 14 b）阶段。最值得注意的是，发育过程的特征在于低M-r α-淀粉酶/胰蛋白酶抑制剂、丝氨酸蛋白酶抑制剂和参与保护免受氧化应激的酶的积累。我们提出的蛋白质的例子，以前没有实验观察到，巯基结合蛋白质的差异提取，和可能的等位基因特异性斑点的变化。我们的研究结果既证实和扩展的知识，从以前的分析个别蛋白质参与灌浆和成熟。
In monocotyledonous plants, the process of seed development involves the deposition of reserves in the starchy endosperm and development of the embryo and aleurone layer. The final stages of seed development are accompanied by an increase in desiccation tolerance and drying out of the mature seed. We have used two-dimensional gel electrophoresis for a time-resolved study of the changes in proteins that occur during seed development in barley (Hordeum vadgare). About 1,000 low-salt extractable protein spots could be resolved on the two-dimensional gels. Protein spots were divided into six categories according to the timing of appearance or disappearance during the 5-week period of comparison. Nineteen different proteins or protein fragments in 36 selected spots were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry (MS) or nano-electrospray tandem MS/MS. Some proteins were present throughout development (for example, cytosolic malate dehydrogenase), whereas others were associated with the early grain filling (ascorbate peroxidase) or desiccation (Cor14b) stages. Most noticeably, the development process is characterized by an accumulation of low-M-r alpha-amylase/trypsin inhibitors, serine protease inhibitors, and enzymes involved in protection against oxidative stress. We present examples of proteins not previously experimentally observed, differential extractability of thiol-bound proteins, and possible allele-specific spot variation. Our results both confirm and expand on knowledge gained from previous analyses of individual proteins involved in grain filling and maturation.