Department of Biology, University of Pisa, Pisa, Italy.
Department of Biology, University of Naples "Federico II", Naples, Italy.
Plant Physiol Biochem. 2018 Jun;127:88-96. doi: 10.1016/j.plaphy.2018.03.014. Epub 2018 Mar 13.
Although some charophytes (sister group to land plants) have been shown to synthesize phytochelatins (PCs) in response to cadmium (Cd), the functional characterization of their phytochelatin synthase (PCS) is still completely lacking. To investigate the metal response and the presence of PCS in charophytes, we focused on the species Nitella mucronata. A 40 kDa immunoreactive PCS band was revealed in mono-dimensional western blot by using a polyclonal antibody against Arabidopsis thaliana PCS1. In two-dimensional western blot, the putative PCS showed various spots with acidic isoelectric points, presumably originated by post-translational modifications. Given the PCS constitutive expression in N. mucronata, we tested its possible involvement in the homeostasis of metallic micronutrients, using physiological concentrations of iron (Fe) and zinc (Zn), and verified its role in the detoxification of a non-essential metal, such as Cd. Neither in vivo nor in vitro exposure to Zn resulted in PCS activation and PC significant biosynthesis, while Fe(II)/(III) and Cd were able to activate the PCS in vitro, as well as to induce PC accumulation in vivo. While Cd toxicity was evident from electron microscopy observations, the normal morphology of cells and organelles following Fe treatments was preserved. The overall results support a function of PCS and PCs in managing Fe homeostasis in the carophyte N. mucronata.
虽然一些轮藻(与陆地植物具有姐妹群关系)已被证明可以在镉(Cd)胁迫下合成植物螯合肽(PCs),但其植物螯合肽合酶(PCS)的功能特征仍完全缺失。为了研究轮藻对金属的响应和 PCS 的存在,我们将重点放在了发菜属物种 Nitella mucronata 上。使用针对拟南芥 PCS1 的多克隆抗体,在一维 Western blot 中揭示了 40 kDa 的免疫反应性 PCS 带。在二维 Western blot 中,推测 PCS 具有各种酸性等电点的斑点,可能是由翻译后修饰引起的。鉴于 N. mucronata 中 PCS 的组成型表达,我们使用生理浓度的铁(Fe)和锌(Zn)测试了其在金属微量元素稳态中的可能作用,并验证了其在非必需金属如 Cd 的解毒中的作用。体内和体外暴露于 Zn 均未导致 PCS 激活和 PC 大量合成,而 Fe(II)/(III) 和 Cd 能够在体外激活 PCS,并在体内诱导 PC 积累。虽然电子显微镜观察到 Cd 的毒性,但 Fe 处理后细胞和细胞器的正常形态得以保留。总的来说,这些结果支持 PCS 和 PCs 在管理轮藻 N. mucronata 中铁稳态中的功能。