Plant Cell, Tissue and Organ Culture

, Volume 94, Issue 3, pp 291–298 | Cite as

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

  • Vaclav Diopan
  • Violetta Shestivska
  • Vojtech Adam
  • Tomas Macek
  • Martina Mackova
  • Ladislav Havel
  • Rene Kizek
Original Paper

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.

Keywords

Metallothionein Tobacco Nicotiana tabacum Thiols Phytoremediation Electrochemical detection Brdicka reaction Voltammetry Liquid chromatography 

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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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Vaclav Diopan
    • 1
    • 2
  • Violetta Shestivska
    • 1
    • 2
  • Vojtech Adam
    • 2
    • 3
  • Tomas Macek
    • 4
  • Martina Mackova
    • 5
  • Ladislav Havel
    • 1
  • Rene Kizek
    • 2
  1. 1.Department of Plant Biology, Faculty of AgronomyMendel University of Agriculture and ForestryBrnoCzech Republic
  2. 2.Department of Chemistry and Biochemistry, Faculty of AgronomyMendel University of Agriculture and ForestryBrnoCzech Republic
  3. 3.Department of Animal Nutrition and Forage Production, Faculty of AgronomyMendel University of Agriculture and ForestryBrnoCzech Republic
  4. 4.Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech RepublicPraha 6Czech Republic
  5. 5.Institute of Chemical TechnologyPrague 6Czech Republic

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