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基于二维差异凝胶电泳的福岛县饭馆村低剂量伽马辐射下水稻幼苗叶片蛋白质组表达变化

2D-DIGE-based proteome expression changes in leaves of rice seedlings exposed to low-level gamma radiation at Iitate village, Fukushima.

作者信息

Hayashi Gohei, Moro Carlo F, Rohila Jai Singh, Shibato Junko, Kubo Akihiro, Imanaka Tetsuji, Kimura Shinzo, Ozawa Shoji, Fukutani Satoshi, Endo Satoru, Ichikawa Katsuki, Agrawal Ganesh Kumar, Shioda Seiji, Hori Motohide, Fukumoto Manabu, Rakwal Randeep

机构信息

a Research Reactor Institute; Kyoto University ; Osaka , Japan.

b Institute of Development; Aging and Cancer; Tohoku University ; Sendai , Japan.

出版信息

Plant Signal Behav. 2015;10(12):e1103406. doi: 10.1080/15592324.2015.1103406.

Abstract

The present study continues our previous research on investigating the biological effects of low-level gamma radiation in rice at the heavily contaminated Iitate village in Fukushima, by extending the experiments to unraveling the leaf proteome. 14-days-old plants of Japonica rice (Oryza sativa L. cv. Nipponbare) were subjected to gamma radiation level of upto 4 µSv/h, for 72 h. Following exposure, leaf samples were taken from the around 190 µSv/3 d exposed seedling and total proteins were extracted. The gamma irradiated leaf and control leaf (harvested at the start of the experiment) protein lysates were used in a 2-D differential gel electrophoresis (2D-DIGE) experiment using CyDye labeling in order to asses which spots were differentially represented, a novelty of the study. 2D-DIGE analysis revealed 91 spots with significantly different expression between samples (60 positive, 31 negative). MALDI-TOF and TOF/TOF mass spectrometry analyses revealed those as comprising of 59 different proteins (50 up-accumulated, 9 down-accumulated). The identified proteins were subdivided into 10 categories, according to their biological function, which indicated that the majority of the differentially expressed proteins consisted of the general (non-energy) metabolism and stress response categories. Proteome-wide data point to some effects of low-level gamma radiation exposure on the metabolism of rice leaves.

摘要

本研究延续了我们之前的研究,通过扩展实验以解析叶片蛋白质组,来调查福岛县饭舘村重度污染地区低剂量伽马辐射对水稻的生物学效应。对14日龄的粳稻(日本晴)植株施加高达4微希沃特/小时的伽马辐射,持续72小时。辐照后,从约190微希沃特/3天辐照的幼苗上采集叶片样本,并提取总蛋白。伽马辐照叶片和对照叶片(在实验开始时收获)的蛋白裂解物用于二维差异凝胶电泳(2D-DIGE)实验,采用CyDye标记,以评估哪些斑点有差异表达,这是本研究的一个创新点。2D-DIGE分析显示,样本间有91个斑点表达差异显著(60个上调,31个下调)。基质辅助激光解吸电离飞行时间(MALDI-TOF)和飞行时间串联质谱(TOF/TOF)分析显示,这些斑点包含59种不同蛋白质(50种积累上调,9种积累下调)。根据其生物学功能,将鉴定出的蛋白质分为10类,这表明大多数差异表达蛋白质属于一般(非能量)代谢和应激反应类别。全蛋白质组数据表明低剂量伽马辐射暴露对水稻叶片代谢有一些影响。

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