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ECOTOX ENVIRON SAFE :天津师范大学丨NHX1转基因浮萍的耐镉生理机制(附NMT实验体系)

转自中关村旭月非损伤微测技术产业联盟

 

 

  • 期刊:Ecotoxicology and Environmental Safety
  • 主题:镉胁迫下Na+/H+逆向转运蛋白(NHX1)转基因浮萍中镉的积累减少
  • 标题:Declined cadmium accumulation in Na+/H+ antiporter (NHX1) transgenic duckweed under cadmium stress
  • 影响因子:4.527
  • 检测指标:Cd2+、H+流速
  • 检测部位:浮萍距离根尖100μm
  • Cd2+、H+流速流实验处理方法:浮萍在50μM CdCl2和50μM CdC+0.5mM阿米洛利分别处理2分钟,20分钟和2小时
  • Cd2+、H+流速流实验测试液成份:0.05mM CdCl2, 0.1mM CaCl2, 0.1mM KCl, 0.3mM MES, pH 5.5
  • 通讯作者:天津师范大学生命科学学院孙金生、杨琳

英文摘要

Cadmium (Cd) is a serious threat to plants health. Though some genes have been reported to get involved in the regulation of tolerance to Cd, the mechanisms underlying this process are not fully understood. Na+/H+ antiporter (NHX1) plays an important role in Na+/H+ trafficking. The salt and cadmium stress tolerance were found to be enhanced by NHX1 in duckweed according to our previous study, however, its function in Cd2+ flux under Cd stress has not been studied.

Here we explored the Cd2+ flux in wild type (WT) and NHX1 transgenic duckweed (NHX1) under Cd stress. We found that the Cd2+ influx in NHX1 duckweed was significantly declined, followed by an increased Cd2+ efflux after 20 min treatment of Cd, which resulted a less accumulation of Cd in NHX1.

Reversely, inhibition of NHX1 by amiloride treatment, enhanced Cd2+ influx in NHX1 duckweed, subsequently delayed Cd2+ efflux in both genotypes of duckweed under Cd2+ shock. H+ efflux in NHX1 duckweed was lower compare with that in WT with 20 min Cd2+ shock. NHX1 also increased the pH value with Cd2+ stress in the transgenic rhizoid. These finding suggested a new function of NHX1 in regulation of Cd2+ and H+ flow during short-term Cd2+ shock.

 

中文摘要(谷歌机翻)

镉(Cd)是对植物健康的严重威胁。虽然据报道一些基因参与调节对Cd的耐受性,但这一过程背后的机制尚不完全清楚。 Na+/H+逆向转运蛋白(NHX1)在Na+/H+运输中起重要作用。根据我们之前的研究,浮萍中的NHX1对盐和镉胁迫耐受性有增强作用,但其在Cd胁迫下对Cd2+ 通量的作用尚未得到研究。

在这里我们探讨了Cd胁迫下野生型(WT)和NHX1转基因浮萍(NHX1)中的Cd2+ 通量。我们发现NHX1浮萍中的Cd2+ 流入量显着下降,随后Cd处理20min后Cd2+ 流出量增加,导致NHX1中Cd的积累减少。

相反,阿米洛利处理对NHX1的抑制作用增强了NHX1浮萍中Cd2+ 的流入,延缓了Cd2+ 冲击下浮萍基因型中Cd2+ 的流出。 NHX1浮萍的H+流出量与WT相比,20min Cd2+ 冲击的H+流出量较低。 NHX1也增加了转基因根状茎中Cd2+ 胁迫的pH值。这些发现表明NHX1在短期Cd2+ 休克期间调节Cd2+ 和H+流动的新功能。

 

结果表明:在5分钟的镉处理后,如图a所示,在NHX1浮萍和WT浮萍中均检测到Cd2+吸收。有趣的是,NHX1浮萍中的Cd2+吸收量明显低于WT中的Cd2+吸收量。在5min处理镉时,NHX1浮萍中积累的镉含量较少。 阿米洛利是NHX1的抑制剂,用于研究瞬态Cd2+通量,在CdCl2(50μM)处理下NHX1活性下降。如图b所示,NHX1浮萍中的Cd2+吸收显着增加,与用镉和没有阿米洛利处理的WT中的Cd2+流入大致相同。该结果显示NHX1在短时间镉中在Cd2+流入中的作用。

文章链接:https://www.sciencedirect.com/science/article/pii/S0147651319307183?via%3Dihub