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基于生长和叶绿素荧光诱导指标的金属元素对浮萍的比较植物毒性研究

Comparative Phytotoxicity of Metallic Elements on Duckweed L. Using Growth- and Chlorophyll Fluorescence Induction-Based Endpoints.

作者信息

Irfan Muhammad, Mészáros Ilona, Szabó Sándor, Oláh Viktor

机构信息

Department of Botany, Institute of Biology and Ecology, Faculty of Science and Technology, University of Debrecen, Egyetem Square 1, H-4032 Debrecen, Hungary.

Department of Biology, Institute of Environmental Sciences, University of Nyiregyhaza, H-4401 Nyiregyhaza, Hungary.

出版信息

Plants (Basel). 2024 Jan 12;13(2):215. doi: 10.3390/plants13020215.

Abstract

In this study, we exposed a commonly used duckweed species- L.-to twelve environmentally relevant metals and metalloids under laboratory conditions. The phytotoxic effects were evaluated in a multi-well-plate-based experimental setup by means of the chlorophyll fluorescence imaging method. This technique allowed the simultaneous measuring of the growth and photosynthetic parameters in the same samples. The inhibition of relative growth rates (based on frond number and area) and photochemical efficiency (F/F and Y(II)) were both calculated from the obtained chlorophyll fluorescence images. In the applied test system, growth-inhibition-based phytotoxicity endpoints proved to be more sensitive than chlorophyll-fluorescence-based ones. Frond area growth inhibition was the most responsive parameter with a median EC of 1.75 mg L, while F/F, the more responsive chlorophyll-fluorescence-based endpoint, resulted in a 5.34 mg L median EC for the tested metals. Ag (EC 0.005-1.27 mg L), Hg (EC 0.24-4.87 mg L) and Cu (EC 0.37-1.86 mg L) were the most toxic elements among the tested ones, while As(V) (EC 47.15-132.18 mg L), Cr(III) (EC 6.22-19.92 mg L), Se(VI) (EC 1.73-10.39 mg L) and Zn (EC 3.88-350.56 mg L) were the least toxic ones. The results highlighted that multi-well-plate-based duckweed phytotoxicity assays may reduce space, time and sample volume requirements compared to the standard duckweed growth inhibition tests. These benefits, however, come with lowered test sensitivity. Our multi-well-plate-based test setup resulted in considerably higher median EC (3.21 mg L) for frond-number-based growth inhibition than the 0.683 mg L median EC derived from corresponding data from the literature with standardized -tests. Under strong acute phytotoxicity, frond parts with impaired photochemical functionality may become undetectable by chlorophyll fluorometers. Consequently, the plant parts that are still detectable display a virtually higher average photosynthetic performance, leading to an underestimation of phytotoxicity. Nevertheless, multi-well-plate-based duckweed phytotoxicity assays, combined with chlorophyll fluorescence imaging, offer definite advantages in the rapid screening of large sample series or multiple species/clones. As chlorophyll fluorescence images provide information both on the photochemical performance of the test plants and their morphology, a joint analysis of the two endpoint groups is recommended in multi-well-plate-based duckweed phytotoxicity assays to maximize the information gained from the tests.

摘要

在本研究中,我们在实验室条件下,将一种常用浮萍物种——浮萍(Lemna minor)暴露于12种与环境相关的金属和类金属中。通过基于多孔板的实验装置,采用叶绿素荧光成像法评估其植物毒性效应。该技术能够同时测量同一样本中的生长和光合参数。根据获得的叶绿素荧光图像计算相对生长率(基于叶状体数量和面积)和光化学效率(Fv/Fm和Y(II))的抑制率。在所应用的测试系统中,基于生长抑制的植物毒性终点指标被证明比基于叶绿素荧光的指标更敏感。叶状体面积生长抑制是最敏感的参数,其半数效应浓度(EC)为1.75 mg/L,而基于叶绿素荧光的更敏感终点指标Fv/Fm,其测试金属的半数效应浓度为5.34 mg/L。在所测试的元素中,银(EC为0.005 - 1.27 mg/L)、汞(EC为0.24 - 4.87 mg/L)和铜(EC为0.37 - 1.86 mg/L)是毒性最强的元素,而五价砷(EC为47.15 - 132.18 mg/L)、三价铬(EC为6.22 - 19.92 mg/L)、六价硒(EC为1.73 - 10.39 mg/L)和锌(EC为3.88 - 350.56 mg/L)是毒性最弱的元素。结果表明,与标准浮萍生长抑制试验相比,基于多孔板的浮萍植物毒性试验可能会减少空间、时间和样本量需求。然而,这些好处伴随着测试灵敏度的降低。我们基于多孔板的测试装置得出基于叶状体数量的生长抑制的半数效应浓度中位数(3.21 mg/L),远高于文献中标准化试验相应数据得出的0.683 mg/L的半数效应浓度中位数。在强烈的急性植物毒性作用下,光化学功能受损的叶状体部分可能无法被叶绿素荧光仪检测到。因此,仍可检测到的植物部分实际上显示出更高的平均光合性能,导致对植物毒性的低估。尽管如此,基于多孔板的浮萍植物毒性试验与叶绿素荧光成像相结合,在快速筛选大量样本系列或多个物种/克隆方面具有明显优势。由于叶绿素荧光图像既提供了受试植物的光化学性能信息,也提供了其形态信息,因此在基于多孔板的浮萍植物毒性试验中,建议对这两个终点指标组进行联合分析,以最大限度地从试验中获取信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5e3/10821045/a7493317836f/plants-13-00215-g001.jpg

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