A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf
A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf
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DOI：
10.1002/elps.200500334
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发表时间：
2005-12-01
期刊：
影响因子：
2.9
通讯作者：
Usui, K
中科院分区：
文献类型：
--
作者：
Kim, DW;Rakwal, R;Usui, K
利用水稻水培系统，采用蛋白质组学和经典生化方法研究了模式水稻品种日本晴对盐胁迫的生理生化响应。吉田氏营养液（YS）用于培育水稻幼苗。YS-生长的18天龄幼苗表现出高度稳定和可重复的症状，当将它们转移到含有130 mM NaCl的YS中4天进行盐胁迫时，显著的是第3叶的萎蔫和布朗宁，光合活性降低，总体幼苗生长受到抑制，并且不能发育新的（第5）叶。由于水稻叶片对盐胁迫的反应是蛋白质组学研究最少的，我们使用第3叶作为源材料。比较未处理对照和盐胁迫的第3叶之间的2-DE蛋白质图谱显示55个差异表达的CBB染色斑点，其中47个斑点比对照增加。这些变化的斑点，身份的33个蛋白质斑点（27个增加和5个减少）的nESI-LC-MS/MS确定。这些确定的蛋白质大多属于主要的代谢过程，如光合二氧化碳同化和光呼吸，这表明盐胁迫响应蛋白和叶片形态之间有很好的相关性。此外，2-DE免疫印迹和酶活性分析的第3叶显示显着变化的关键标志酶与氧化损伤盐胁迫：抗坏血酸过氧化物酶和脂质过氧化反应诱导，过氧化氢酶被抑制。这些结果表明，水培体系最适合于水稻幼苗盐胁迫蛋白质组学研究。
By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day-old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling.