Ma Jie, Sheng Huachun, Li Xiuli, Wang Lijun
College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China.
College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China.
Plant Physiol Biochem. 2016 Jul;104:71-80. doi: 10.1016/j.plaphy.2016.03.024. Epub 2016 Mar 19.
Silicon (Si) can alleviate cadmium (Cd) stress in rice (Oryza sativa) plants, however, the understanding of the molecular mechanisms at the single-cell level remains limited. To address these questions, we investigated suspension cells of rice cultured in the dark environment in the absence and presence of Si with either short- (12 h) or long-term (5 d) Cd treatments using a combination of isobaric tags for relative and absolute quantitation (iTRAQ), fluorescent staining, and inductively coupled plasma mass spectroscopy (ICP-MS). We identified 100 proteins differentially regulated by Si under the short- or long-term Cd stress. 70% of these proteins were down-regulated, suggesting that Si may improve protein use efficiency by maintaining cells in the normal physiological status. Furthermore, we showed two different mechanisms for Si-mediated Cd tolerance. Under the short-term Cd stress, the Si-modified cell walls inhibited the uptake of Cd ions into cells and consequently reduced the expressions of glycosidase, cell surface non-specific lipid-transfer proteins (nsLTPs), and several stress-related proteins. Under the long-term Cd stress, the amount of Cd in the cytoplasm in Si-accumulating (+Si) cells was decreased by compartmentation of Cd into vacuoles, thus leading to a lower expression of glutathione S-transferases (GST). These results provide protein-level insights into the Si-mediated Cd detoxification in rice single cells.
硅(Si)可缓解水稻(Oryza sativa)植株的镉(Cd)胁迫,然而,在单细胞水平上对其分子机制的了解仍然有限。为了解决这些问题,我们使用相对和绝对定量的等压标签(iTRAQ)、荧光染色和电感耦合等离子体质谱(ICP-MS)相结合的方法,研究了在黑暗环境中培养的水稻悬浮细胞,这些细胞在有无硅的情况下分别接受短期(12小时)或长期(5天)的镉处理。我们鉴定出100种在短期或长期镉胁迫下受硅差异调节的蛋白质。其中70%的蛋白质表达下调,这表明硅可能通过维持细胞的正常生理状态来提高蛋白质利用效率。此外,我们还展示了硅介导的镉耐受性的两种不同机制。在短期镉胁迫下,硅修饰的细胞壁抑制了镉离子进入细胞,从而降低了糖苷酶、细胞表面非特异性脂质转移蛋白(nsLTPs)和几种胁迫相关蛋白的表达。在长期镉胁迫下,通过将镉分隔到液泡中,硅积累(+Si)细胞细胞质中的镉含量降低,从而导致谷胱甘肽S-转移酶(GST)的表达降低。这些结果为水稻单细胞中硅介导的镉解毒提供了蛋白质水平的见解。