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两种油菜(甘蓝型油菜)品种叶片中铬胁迫和硫缺乏响应的蛋白质组学分析,这两个品种对六价铬的耐受性存在差异。

Proteomic analysis of chromium stress and sulfur deficiency responses in leaves of two canola (Brassica napus L.) cultivars differing in Cr(VI) tolerance.

机构信息

Department of Molecular Biology and Genetics, Faculty of Science and Literature, Afyon Kocatepe University, 03200 Afyonkarahisar, Turkey.

Department of Molecular Biology and Genetics, Faculty of Science and Literature, Afyon Kocatepe University, 03200 Afyonkarahisar, Turkey.

出版信息

Ecotoxicol Environ Saf. 2016 Feb;124:255-266. doi: 10.1016/j.ecoenv.2015.10.023. Epub 2015 Nov 5.

Abstract

Sulfur (S) is an essential macronutrient for plant growth and development, and it plays an essential role in response to environmental stresses. Plants suffer with combined stress of S deficiency and hexavalent chromium [Cr(VI)] in the rhizosphere. Little is known about the impact of S deficiency on leaf metabolism of canola (Brassica napus L.) under Cr(VI) stress. Therefore, this study is the first to examine the effects of Cr(VI) stress and S deficiency in canola at a molecular level. A comparative proteomic approach was used to investigate the differences in protein abundance between Cr-tolerant NK Petrol and Cr-sensitive Sary cultivars. The germinated seeds were grown hydroponically in S-sufficient (+S) nutrient solution for 7 days and then subjected to S-deficiency (-S) for 7 days. S-deficient and +S seedlings were then exposed to 100μM Cr(VI) for 3 days. Protein patterns analyzed by two-dimensional electrophoresis (2-DE) revealed that 58 protein spots were differentially regulated by Cr(VI) stress (+S/+Cr), S-deficiency (-S/-Cr) and combined stress (-S/+Cr). Of these, 39 protein spots were identified by MALDI-TOF/TOF mass spectrometry. Differentially regulated proteins predominantly had functions not only in photosynthesis, but also in energy metabolism, stress defense, protein folding and stabilization, signal transduction, redox regulation and sulfur metabolism. Six stress defense related proteins including 2-Cys peroxiredoxin BAS1, glutathione S-transferase, ferritin-1, l-ascorbate peroxidase, thiazole biosynthetic enzyme and myrosinase-binding protein-like At3g16470 exhibited a greater increase in NK Petrol. The stress-related proteins play an important role in the detoxification of Cr(VI) and maintaining cellular homeostasis under variable S nutrition.

摘要

硫(S)是植物生长和发育所必需的大量营养素,它在应对环境胁迫方面起着至关重要的作用。植物在根际受到 S 缺乏和六价铬 [Cr(VI)] 的联合胁迫。关于 S 缺乏对油菜(Brassica napus L.)叶片代谢在 Cr(VI)胁迫下的影响知之甚少。因此,本研究首次在分子水平上研究了 Cr(VI)胁迫和 S 缺乏对油菜的影响。采用比较蛋白质组学方法研究了耐铬 NK Petrol 和敏感 Sary 品种之间蛋白质丰度的差异。将发芽的种子在 S 充足(+S)营养液中水培培养 7 天,然后进行 S 缺乏(-S)培养 7 天。然后,将 S 缺乏和+S 幼苗暴露于 100μM Cr(VI)中 3 天。通过二维电泳(2-DE)分析的蛋白质图谱显示,58 个蛋白质斑点受 Cr(VI)胁迫(+S/+Cr)、S 缺乏(-S/-Cr)和联合胁迫(-S/+Cr)调节。其中,39 个蛋白质斑点通过 MALDI-TOF/TOF 质谱鉴定。差异调节的蛋白质主要具有不仅在光合作用中,而且在能量代谢、应激防御、蛋白质折叠和稳定、信号转导、氧化还原调节和硫代谢中具有功能。包括 2-Cys 过氧化物酶 BAS1、谷胱甘肽 S-转移酶、铁蛋白-1、l-抗坏血酸过氧化物酶、噻唑生物合成酶和拟南芥结合蛋白样 At3g16470 在内的 6 种应激防御相关蛋白在 NK Petrol 中表现出更大的增加。应激相关蛋白在可变 S 营养条件下对 Cr(VI)的解毒和维持细胞内稳态中起着重要作用。

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