Institute of Environmental Engineering, Faculty of Infrastructure and Environment, Czestochowa University of Technology, Czestochowa, Poland.
Laboratory of Civil Engineering and Environment (LGCgE), Environmental Axis, University of Lille, F-59650, Villeneuve d'Ascq, France.
Environ Sci Pollut Res Int. 2019 Jun;26(16):16040-16052. doi: 10.1007/s11356-019-04972-y. Epub 2019 Apr 9.
Yellow-lupin (Lupinus luteus L.) was grown on soils contaminated with heavy metals during two parallel studies. In the first one, the soil was contaminated by industrial activities whereas, in the second one, the soil was artificially contaminated with a single metal including Cd, Pb, Zn, Ni (in nitrate form), and Ag (in nitrate and nanoparticles form). The study was performed to assess a plant's response to contamination including its antioxidative response and molecular mechanisms involved in metal detoxification through the expression level of metallothioneins (MTs). Overall, the study provided insights into identification and validation of housekeeping genes (HKG) in L. luteus under exposure to metal stress and showed the effects of selected heavy metals and silver nanoparticles on the expression of metallothioneins, the activity of guaiacol peroxidase (GPX) and bioaccumulation of metals in leaves of L. luteus. As such, HKG validation using BestKeeper, NormFinder, and geNorm software allowed for the selection of four most stable reference genes in a context metal contamination for the selected plant. Moreover, a significant increase in the expression levels of MT was observed in plants grown under heavy metal stress and none on plants grown on 25 mg kg of silver nanoparticles. Also, the GPX activity and MT expression showed statistically significant changes between different conditions and doses which means that they can be used as highly sensitive stress markers for planning the phytoremediation process on a large scale.
在两项平行研究中，种植了在重金属污染土壤上生长的黄羽扇豆（ Lupinus luteus L.）。在第一项研究中，土壤受到工业活动的污染，而在第二项研究中，土壤被人为地用单一金属（包括 Cd、Pb、Zn、Ni（硝酸盐形式）和 Ag（硝酸盐和纳米颗粒形式））污染。这项研究旨在评估植物对污染的反应，包括其抗氧化反应和参与金属解毒的分子机制，方法是通过金属硫蛋白（MTs）的表达水平来评估。总的来说，该研究提供了在金属胁迫下识别和验证黄羽扇豆看家基因（HKG）的见解，并展示了选定重金属和银纳米颗粒对金属硫蛋白表达、愈创木酚过氧化物酶（GPX）活性和黄羽扇豆叶片金属生物累积的影响。因此，使用 BestKeeper、NormFinder 和 geNorm 软件对 HKG 进行验证，允许在选定植物的金属污染背景下选择四个最稳定的参考基因。此外，在重金属胁迫下生长的植物中观察到 MT 表达水平显著增加，而在生长在 25mgkg 银纳米颗粒下的植物中则没有增加。此外，GPX 活性和 MT 表达在不同条件和剂量之间表现出统计学上的显著变化，这意味着它们可以作为高度敏感的应激标志物，用于规划大规模的植物修复过程。
