Laboratory of Plant Biotechnology, Faculty of Sciences, BP 1171, 3000 Sfax, University of Sfax, Tunisia; Laboratoire de Génie Civil et géo-Environnement, Université de Lille 1, F-59655 Villeneuve d'Ascq, France.
Laboratoire de Génie Civil et géo-Environnement, Université de Lille 1, F-59655 Villeneuve d'Ascq, France.
Chemosphere. 2018 Apr;197:123-134. doi: 10.1016/j.chemosphere.2017.12.146. Epub 2017 Dec 28.
Phytochelatin synthase and metallothionein gene expressions were monitored via qPCR in order to investigate the molecular mechanisms involved in Cd and Cr detoxification in date palm (Phoenix dactylifera). A specific reference gene validation procedure using BestKeeper, NormFinder and geNorm programs allowed selection of the three most stable reference genes in a context of Cd or Cr contamination among six reference gene candidates, namely elongation factor α1, actin, aldehyde dehydrogenase, SAND family, tubulin 6 and TaTa box binding protein. Phytochelatin synthase (pcs) and metallothionein (mt) encoding gene expression were induced from the first days of exposure. At low Cd stress (0.02 mM), genes were still up-regulated until 60th day of exposure. At the highest metal concentrations, however, pcs and mt gene expressions decreased. pcs encoding gene was significantly up-regulated under Cr exposure, and was more responsive to increasing Cr concentration than mt encoding gene. Moreover, exposure to Cd or Cr influenced clearly seed germination and hypocotyls elongation. Thus, the results have proved that both analyzed genes participate in metal detoxification and their expression is regulated at transcriptional level in date palm subjected to Cr and Cd stress. Consequently, variations of expression of mt and pcs genes may serve as early-warning biomarkers of metal stress in this species.
为了研究在刺梨(Phoenix dactylifera)中 Cd 和 Cr 解毒所涉及的分子机制,通过 qPCR 监测植物螯合肽合酶和金属硫蛋白基因的表达。使用 BestKeeper、NormFinder 和 geNorm 程序进行特定的参考基因验证程序,允许在 Cd 或 Cr 污染的情况下,从六个候选参考基因中选择三个最稳定的参考基因,即延伸因子α1、肌动蛋白、醛脱氢酶、SAND 家族、微管蛋白 6 和 TaTa 框结合蛋白。植物螯合肽合酶(pcs)和金属硫蛋白(mt)编码基因的表达在暴露的第一天就被诱导。在低 Cd 胁迫(0.02 mM)下,基因仍被上调至暴露 60 天。然而,在最高金属浓度下,pcs 和 mt 基因的表达下降。Cr 暴露下 pcs 编码基因显著上调,对 Cr 浓度增加的反应比对 mt 编码基因更为敏感。此外,Cd 或 Cr 的暴露明显影响种子发芽和下胚轴伸长。因此,研究结果证明,这两个分析基因都参与了金属解毒,并且在受到 Cr 和 Cd 胁迫的刺梨中,其表达在转录水平上受到调控。因此,mt 和 pcs 基因表达的变化可以作为该物种金属胁迫的早期预警生物标志物。