Proteomic analysis of rice seedlings during cold stress
Proteomic analysis of rice seedlings during cold stress
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DOI：
10.1002/pmic.200600921
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发表时间：
2007-04-01
期刊：
影响因子：
3.4
通讯作者：
Komatsu, Setsuko
中科院分区：
文献类型：
--
作者：
Hashimoto, Makoto;Komatsu, Setsuko
低温是影响植物生长和农业生产的重要环境变化之一。为了研究水稻对冷胁迫的反应，利用蛋白质组学方法分析了蛋白质表达的变化。以2周龄的水稻幼苗为材料，在5 ℃下处理48 h，提取叶片、叶鞘和根的总蛋白，经双向电泳分离，CBB染色。在每个器官的250-400个蛋白质点中，有39个蛋白质在冷胁迫后发生了丰度变化，其中19个蛋白质含量增加，20个蛋白质含量减少。在叶片中，由于存在丰富的二磷酸核酮糖羧化酶/加氧酶大亚基（RuBisCO LSU）蛋白，约占总蛋白的50%，因此很难检测到胁迫反应蛋白的变化。为了克服这个问题，制备抗体亲和柱以捕获RuBisCO LSU，随后使用2-DE分离来自柱的流通液中的剩余蛋白质。结果表明，胁迫反应蛋白的变化很小，在冷胁迫后新检测到4种蛋白。从鉴定的蛋白质中可以看出，低温胁迫使叶片中与能量代谢相关的蛋白质表达上调，而与防御相关的蛋白质表达下调。这些结果表明，能量生产被激活的低温环境中，此外，应激相关蛋白迅速上调，而防御相关蛋白消失，在长期的冷胁迫。
Low temperature is one of the important environmental changes that affect plant growth and agricultural production. To investigate the responses of rice to cold stress, changes in protein expression were analyzed using a proteomic approach. Two-week-old rice seedlings were exposed to 5 degrees C for 48 h, then total crude proteins were extracted from leaf blades, leaf sheaths and roots, separated by 2-DE and stained with CBB. Of the 250-400 protein spots from each organ, 39 proteins changed in abundance after cold stress, with 19 proteins increasing, and 20 proteins decreasing. In leaf blades, it was difficult to detect the changes in stress-responsive proteins due to the presence of an abundant protein, ribulose bisphosphate carboxylase/oxygenase large subunit (RuBisCO LSU), which accounted for about 50% of the total proteins. To overcome this problem, an antibody-affinity column was prepared to trap RuBisCO LSU, and the remaining proteins in the flow through from the column were subsequently separated using 2-DE. As a result, slight changes in stress responsive proteins were clearly displayed, and four proteins were newly detected after cold stress. From identified proteins, it was concluded that proteins related to energy metabolism were up-regulated, and defense-related proteins were down-regulated in leaf blades, by cold stress. These results suggest that energy production is activated in the chilling environment; furthermore, stress-related proteins are rapidly up-regulated, while defense-related proteins disappear, under long-term cold stress.