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Determination of content of metallothionein and low molecular mass stress peptides in transgenic tobacco plants

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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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Abbreviations

MT:

Metallothionein

DPV:

Differential pulse voltammetry

AdTS:

Adsorptive transfer stripping

PC:

Phytochelatins

Cys :

Cysteine

GSH:

Reduced glutathione

GSSG:

Oxidised glutathione

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Acknowledgements

The authors wish to express their thanks to Dr. Grace J. Chavis for language corrections and discussions. The financial support from grant No. 1M06030 is greatly acknowledged.

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Authors and Affiliations

  1. Department of Plant Biology, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic

    Vaclav Diopan, Violetta Shestivska & Ladislav Havel

  2. Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic

    Vaclav Diopan, Violetta Shestivska, Vojtech Adam & Rene Kizek

  3. Department of Animal Nutrition and Forage Production, Faculty of Agronomy, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00, Brno, Czech Republic

    Vojtech Adam

  4. Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo n. 2, 166 10, Praha 6, Czech Republic

    Tomas Macek

  5. Institute of Chemical Technology, Technicka 5, 166 28, Prague 6, Czech Republic

    Martina Mackova

Authors
  1. Vaclav Diopan
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  2. Violetta Shestivska
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  3. Vojtech Adam
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  4. Tomas Macek
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  5. Martina Mackova
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  6. Ladislav Havel
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  7. Rene Kizek
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Correspondence to Rene Kizek.

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Diopan, V., Shestivska, V., Adam, V. et al. Determination of content of metallothionein and low molecular mass stress peptides in transgenic tobacco plants. Plant Cell Tiss Organ Cult 94, 291–298 (2008). https://doi.org/10.1007/s11240-008-9356-2

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  • DOI: https://doi.org/10.1007/s11240-008-9356-2

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