Determination of content of metallothionein and low molecular mass stress peptides in transgenic tobacco plants |
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Authors: | Vaclav Diopan Violetta Shestivska Vojtech Adam Tomas Macek Martina Mackova Ladislav Havel Rene Kizek |
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Institution: | 1. Department of Plant Biology, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic 2. Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic 3. Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic 4. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 166 10, Praha 6, Czech Republic 5. Institute of Chemical Technology, Technicka 5, 166 28, Prague 6, Czech Republic
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Abstract: | Phytoremediation is a process that utilizes plants to remove, transfer, stabilize, or destroy pollutants in soil, sediment,
and groundwater. Plants used for such purposes have several requirements. Genetic engineering these plants could be an effective
tool used to acquire features needed for such purposes within a substantial amount of time. This paper aims to utilize electrochemical
techniques to analyze transgenic tobacco and, thus, to reveal their heavy metals phytoremediation potential. Total thiol and
metallothionein (MT) quantities were determined in the control and transgenic tobacco plants. The total content of thiols
in transgenic plants varied within the range of 561 to 1,671 μg g−1. Furthermore, the determination of MT was done on transgenic tobacco plants. The level of human MT in transgenic tobacco
plants varied between 25 and 95 μg g−1. However, a plant cell protects itself by synthesizing low molecular mass thiols such as reduced glutathione and phytochelatins
to protect itself against heavy metals toxicity. The most important thiols, cysteine (Cys), glutathione (GSH), oxidised glutathione
(GSSG) and phytochelatin 2 (PC2), were determined in the non-transgenic and transgenic tobacco plants by high performance
liquid chromatography with electrochemical detection. Tobacco plants synthesizing the highest amount of metallothionein have
the highest basal level of phytochelatin 2 as well as reduced glutathione and free cysteine. It clearly follows from the results
obtained that the biosynthesis of particular thiols is mutually linked, which contributes to a better protection of a transgenic
plant against heavy metals effects. |
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Keywords: | Metallothionein Tobacco Nicotiana tabacum Thiols Phytoremediation Electrochemical detection Brdicka reaction Voltammetry Liquid chromatography |
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