Spectrometry, Sample Preparation and Mechanization Group-GEPAM, Institute of Chemistry, University of Campinas-Unicamp, P.O. Box 6154, 13083-970, Campinas, SP, Brazil; National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, University of Campinas-Unicamp, P.O. Box 6154, 13083-970, Campinas, SP, Brazil.
Department of Genetics, Laboratory of Biochemistry and Genetics of Plants, Escola Superior de Agricultura Luiz de Queiroz, ESALQ-University of São Paulo, 13400-970, Piracicaba, SP, Brazil.
J Trace Elem Med Biol. 2017 Dec;44:50-58. doi: 10.1016/j.jtemb.2017.05.010. Epub 2017 Jun 3.
The mitigation of Cd-stress through Si addition to Arabidopsis thaliana cultivation is evaluated in terms of total metal content, proteomic and enzymatic approaches. Four different treatment are evaluated: TC (control, without Si or Cd addition), T1 (with Si addition), T2 (with Cd addition), and T3 (with Si and Cd addition). Through the total determination of Cd and Si in Arabidopsis leaves, the Cd concentration decreased by half when T2 is compared with T3 treatment. In terms of proteomic approach, some differential protein species are achieved by comparative proteomics through 2-D DIGE of all treatments evaluated. Fifty six differential abundant proteins spots (abundance factor ≥1.5) are detected, and 32 of them accurately characterized and identified through nESI-LC-MS/MS. These proteins are differentially produced due to Cd and/or Si treatments, which mainly include proteins associated with disease/defense, energy and metabolism. The most difference in the abundance of proteins is found due to the presence or absence of Si in plants treated with Cd. Regarding the enzymatic approaches, a major increase is found on APX, CAT and GR activities (5.0, 3.5, and 1.5-fold, respectively). The same is observed for the MDA concentration because an increase of 3-fold is found when TC are compared to those treated with T2. However, when T3 plants are evaluated, the enzymes activities are similar to TC plants. Differences ranging from 6.5 to 21% are detected considering the activity of SOD in the treatments (T1-T3 x TC). The decreased activities of CAT, APX and GR and lower MDA concentration indicate a lower reactive oxygen species production in plants treated with Cd and Si. Based on a proteomics point of view it is possible to conclude that Si-Cd interactions occur at protein level and allow plants to respond effectively to the Cd toxicity, revealing the active involvement of Si on mechanisms involved in Si-induced Cd tolerance in Arabidopsis plants. Additionally, from an enzymatic point of view, it is possible to conclude that Si positively interferes diminishing the negative effects of Cd in Arabidopsis by decreasing the reactive oxygen species generation and increasing the antioxidative enzyme activity.
通过向拟南芥培养物中添加硅来减轻 Cd 胁迫,从总金属含量、蛋白质组学和酶学方法进行评估。评估了四种不同的处理方法:TC(对照,不添加 Si 或 Cd)、T1(添加 Si)、T2(添加 Cd)和 T3(添加 Si 和 Cd)。通过对拟南芥叶片中 Cd 和 Si 的总测定,与 T3 处理相比,T2 处理时 Cd 浓度降低了一半。在蛋白质组学方法方面,通过对所有评估处理的 2-D DIGE 进行比较蛋白质组学,获得了一些差异蛋白种类。检测到 56 个差异丰度蛋白斑点(丰度因子≥1.5),并通过 nESI-LC-MS/MS 准确鉴定了 32 个蛋白。这些蛋白因 Cd 和/或 Si 处理而差异产生,主要包括与疾病/防御、能量和代谢相关的蛋白。在 Cd 处理的植物中,由于 Si 的存在或不存在,蛋白丰度的差异最大。关于酶学方法,APX、CAT 和 GR 活性(分别增加 5.0、3.5 和 1.5 倍)显著增加。当 TC 与 T2 处理相比时,MDA 浓度也发现增加了 3 倍。然而,当评估 T3 植物时,酶活性与 TC 植物相似。考虑到处理(T1-T3 x TC)中 SOD 的活性,检测到差异率在 6.5%至 21%之间。CAT、APX 和 GR 活性降低和 MDA 浓度降低表明,在添加 Cd 和 Si 的植物中,活性氧的产生减少。从蛋白质组学的角度来看,可以得出结论,Si-Cd 相互作用发生在蛋白质水平上,使植物能够有效应对 Cd 毒性,表明 Si 积极参与了拟南芥植物中 Si 诱导的 Cd 耐受性相关机制。此外,从酶学的角度来看,可以得出结论,Si 通过减少活性氧的产生和增加抗氧化酶的活性,对 Cd 在拟南芥中的负面影响产生积极干扰。