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Lichtscheidl-Schultz, Irene
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document
Response of Three Different Bryophyte Species to Cu and Zn Treatment
Abstract
Bryophytes are non-vascular, multicellular land plants and inhabit extremely different habitats, ranging from temporary dry river banks (e.g. <i>Physcomitrella patens</i>) to metal contaminated sites (e.g. <i>Pohlia drummondii</i> and <i>Mielichhoferia elongata</i>). Therefore, some bryophyte species are considered stress tolerant, and even the supposedly metal sensitive moss<i> P. patens</i> showed increased tolerance to Cu-EDTA in earlier studies [1].In this study, the bryophytes were cultivated on sterile agar plates and tested for zinc (as Zn-EDTA, ZnCl<sub>2</sub> and ZnSO<sub>4</sub>) and copper (as Cu-EDTA, CuCl<sub>2</sub> and CuSO<sub>4</sub>) over a period of five weeks.Despite of the high tolerance towards Cu-EDTA of P. patens, we measured changes in growth and metal uptake by X-ray microanalysis (EDX) in a scanning electron microscope if the metal is offered with different anions. Here, especially the uptake of EDTA chelated metals was significantly lower compared to metal offered as ionic salt. Modelling of ion availability using Visual MINTEQ [2] explained most of the differences in toxicity. In addition to EDX, the fluorescent dyes FluoZin-3 for zinc and Phen Green SK for copper were used to ensure intercellular metal uptake.Reactive oxygen species (ROS) are a general indicator for stress and changes in the cellular content of ROS after staining with 2,7-dichlorofluorescein diacetate (H2DCFDA) [3] were analyzed and the three different bryophyte species compared. <i>P. patens</i> showed only low H<sub>2</sub>DCFDA fluorescence in control cells, in contrast to metal treated cells were increased ROS could be detected for chloroplast associated mitochondria, the nuclear region and the cell wall region.Further investigation of cellular localization of metal deposition was performed using FluoZin-3 and is ongoing in transmission electron microscopy studies.<br/><br/> [1] Sassmann, S., Wernitznig, S., Lichtscheidl, I.K., and Lang, I., 2010, Protoplasma, 246(1-4), 119-123 <br/>[2] Gustafsson, J.P., 2010, KTH Royal Inst. of Technol. Stockholm <br/>[3] Darehshoure, A., and Lütz-Meindl, U., 2010, Protoplasma, 239(1-4), P. 49-56This research was supported by the Vienna Anniversary Foundation for Higher Education (grant H-2486/2012 to S.S.) and the ŒAD (Appear-43/BIOREM).