Separation of proteins from stressed rice (Oryza sativa L.) leaf tissues by two‐dimensional polyacrylamide gel electrophoresis: Induction of pathogenesis‐related and cellular protectant proteins by jasmonic acid, UV irradiation and copper chloride
Separation of proteins from stressed rice (Oryza sativa L.) leaf tissues by two‐dimensional polyacrylamide gel electrophoresis: Induction of pathogenesis‐related and cellular protectant proteins by jasmonic acid, UV irradiation and copper chloride
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DOI：
10.1002/(sici)1522-2683(19991101)20:17
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发表时间：
1999-11
期刊：
影响因子：
2.9
通讯作者：
R. Rakwal;G. Agrawal;M. Yonekura
中科院分区：
文献类型：
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作者：
R. Rakwal;G. Agrawal;M. Yonekura
我们使用了三种胁迫，包括信号化合物茉莉酸，环境胁迫，紫外线照射，和重金属盐氯化铜，研究蛋白质模式的变化在水稻（Oryza sativa L.）叶组织进行双向聚丙烯酰胺凝胶电泳。然而，代替使用含有尿素的裂解缓冲液（O ′ Farrell，J.Biol.Chem.1975，250，4007-4021）从水稻幼苗组织中提取蛋白质，我们使用Tris-HCl缓冲液（通常用于提取蛋白质以通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离）提取蛋白质，并通过O ′ Farrell的常规方法解析这些提取的蛋白质。此外，与对照相比，清楚地观察到大量蛋白质的诱导。在对照实验中没有发现与这些诱导蛋白相对应的斑点，表明在各种胁迫处理后蛋白质模式发生了质的变化。13种蛋白质中的12种可以从茉莉酸处理的水稻叶组织中进行N末端测序，并且一种蛋白质从UV照射的叶组织中测序。这些蛋白质显示出与来自植物（包括水稻）的致病相关蛋白（索马甜样蛋白、PR 5类蛋白、β-1，3-葡聚糖酶前体、由PBZ 1基因编码的细胞内PR蛋白和抗真菌蛋白）和细胞保护剂（谷胱甘肽转移酶，EC 2.5.1.18;和抗坏血酸过氧化物酶）蛋白的高度同源性。这里提出的结果表明茉莉酸在水稻植物的自我防御机制中的作用。
We have used three kinds of stresses, including the signaling compound jasmonic acid, an environmental stressor, UV irradiation, and a heavy metal salt copper chloride, to study changes in the protein patterns in rice (Oryza sativa L.) leaf tissues using two‐dimensional polyacrylamide gel electrophoresis. However, instead of using lysis buffer containing urea (O'Farrell, J. Biol. Chem. 1975, 250, 4007—4021) for extraction of proteins from rice seedling tissues, we used Tris‐HCl buffer (commonly used for extraction of proteins for separation by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis) for extraction of proteins and resolved these extracted proteins by the usual method of O'Farrell. Furthermore, the induction of a large number of proteins was clearly observed over controls. No spots corresponding to these induced proteins were found in the control experiment, indicating qualitative changes in protein patterns after various stress treatments. A total of 12 out of 13 proteins could be N‐terminally sequenced from jasmonic acid‐treated rice leaf tissues, and one protein was sequenced from UV‐irradiated leaf tissues. These proteins showed high homology to pathogenesis‐related (thaumatin‐like protein, a PR5 class protein; a beta‐1,3‐glucanase precursor; an intracellular PR protein encoded by PBZ1 gene, and an antifungal protein) and cellular protectant (glutathione transferase, EC 2.5.1.18; and ascorbate peroxidase) proteins, from plants, including rice. Results presented here suggest a role for jasmonic acid in the self‐defense mechanisms of rice plants.