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 sativusL.)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100μmol·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(100μmol·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。
[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 junol.mZ.s1 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 solution Ⅰ. The extracts of solution Ⅱ and 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 earboxylase. [ 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.
出处
《中国农业科学》
CAS
CSCD
北大核心
2009年第2期588-596,共9页
Scientia Agricultura Sinica
基金
作物生物学国家重点实验室开放课题(200509)
