| 低温对不同基因型黄瓜叶片蛋白质组的影响 |
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| 引用本文： | 许培磊,白吉刚,王秀娟,宗成顺,田明.低温对不同基因型黄瓜叶片蛋白质组的影响[J].中国农业科学,2009,42(2):588-596. |
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| 作者姓名： | 许培磊 白吉刚 王秀娟 宗成顺 田明 |
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| 作者单位： | 1. 山东农业大学生命科学学院/作物生物学国家重点实验室,山东,泰安,271018
2. 山东,烟台莱州市农业局,山东,烟台,261400 |
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| 基金项目： | 作物生物学国家重点实验室开放课题 |
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| 摘 要： | 【目的】分离不同基因型黄瓜叶片的低温诱导蛋白，为从蛋白质组角度揭示黄瓜应答低温逆境的调控机制打下基础。【方法】以黄瓜（Cucumis sativus L.）的日光温室品种‘新泰密刺’和露地栽培品种‘津研4号’为试材，采用分步提取法溶解叶片总蛋白质，运用SDS聚丙烯酰胺凝胶电泳、双向电泳、质谱分析与检索技术，在100 μmol?m-2?s-1的光照条件下，比较研究低温（15/15℃）和常温（25/18℃）下叶片蛋白质组的差异。【结果】利用提取液Ⅰ、Ⅱ和Ⅲ分步提取叶片总蛋白质后，提取液Ⅰ溶解的黄瓜蛋白质较少。对提取液Ⅱ和Ⅲ溶解的叶片蛋白质进行双向电泳后发现，相对于常温处理，低温使‘新泰密刺’的7个蛋白质点特异表达、2个蛋白质点上调表达、2个蛋白质点下调表达、5个蛋白质点未表达;使‘津研4号’的7个蛋白质点特异表达、1个蛋白质点上调表达、4个蛋白质点下调表达、9个蛋白质点未表达。低温下，‘新泰密刺’的蛋白质点3和4 以及‘津研4号’中蛋白质点2的表达量显著高于常温处理，通过MALDI-TOF质谱分析，这3个蛋白质点被鉴定为1,5-二磷酸核酮糖羧化酶的大亚基。【结论】黄瓜叶片的高亲水性蛋白质含量低;在相同的光照（100 μmol?m-2?s-1）条件下低温对不同基因型黄瓜叶片的蛋白质组均有影响;1,5-二磷酸核酮糖羧化酶可能与黄瓜叶片耐低温有关。
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| 关 键 词： | 黄瓜 蛋白质组 低温 双向电泳 质谱分析 5-二磷酸核酮糖羧化酶 |
| 收稿时间： | 2008-1-30 |
Effects of Low Temperature on Leaf Proteome from Cucumis sativus of Different Genotypes |
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| XU Pei-lei,BAI Ji-gang,WANG Xiu-juan,ZONG Cheng-shun,TIAN Ming.Effects of Low Temperature on Leaf Proteome from Cucumis sativus of Different Genotypes[J].Scientia Agricultura Sinica,2009,42(2):588-596. |
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| Authors: | XU Pei-lei BAI Ji-gang WANG Xiu-juan ZONG Cheng-shun TIAN Ming |
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| Institution: | State Key Laboratory of Crop Biology/College of Life Sciences, Shandong Agricultural University |
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| Abstract: | 【Objective】 The purpose of this project was to separate protein spots induced by low temperature in cucumber leaves of two genotypes, and reveal the mechanism of cucumber responding to low temperature. 【Method】Two cucumber genotypes (Cucumis sativus L. cv Xintaimici and Jinyan 4) were both treated with low temperature (15/15℃) and normal temperature (25/18℃) under the light of 100 μmol?m-2?s-1 for 7 d to study the changes of leaf proteome. The total proteins were extracted by three-steps, separated by two-dimensional gel electrophoresis (2-DE) and analysed through MALDI-TOF mass spectrometry. 【Result】 SDS-PAGE showed that few proteins were extracted by hydrophilic solutionⅠ. The extracts of solution Ⅱand Ⅲ were separated through 2-DE. Compared with treatment of normal temperature, low temperature made ‘Xintaimici’ genotype to produce 7 special protein spots, 2 up-regulated protein spots, 2 down-regulated protein spots and 5 unexpressed protein spots, and allowed ‘Jinyan 4’ to produce 7 special protein spots, 1 up-regulated protein spot, 4 down-regulated protein spots and 9 unexpressed protein spots. Three special protein spots, which were highly expressed under low temperature, were identified to be the large subunit of ribulose 1,5-bisphosphate carboxylase. 【Conclusion】There were few hydrophilic proteins in cucumber leaves. Low temperature influenced leaf proteome in two cucumber genotypes and different cultivars expressed different proteins. Ribulose 1,5-bisphosphate carboxylase may be related to low-temperature tolerance in cucumber leaves. |
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| Keywords: | Cucumis sativus L proteome low temperature 2-DE MALDI-TOF-MS ribulose 1 5-bisphosphate carboxylase |
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