Skip to main content
Log in

Induced DNA Damage Measured by the Comet Assay in 10 Weed Species

  • Published:
Biologia Plantarum

Abstract

For most plant species growing in polluted areas no mutagenicity assays are available. We have studied the possibility of using the alkaline protocol of the Comet assay as a method for detecting induced DNA damage in wildly growing weeds. The monofuctional alkylating agent ethyl methanesulphonate (EMS) was applied on leaves of 10 weed species (ordered according to the diameter of the nuclei): Arabidopsis thaliana, Convolvulus arvensis, Bellis perennis, Urtica dioica, Lamium album, Chenopodium rubrum, Plantago media, Poa annua, Taraxacum officinale, and Agropyron repens. With increasing concentrations of EMS (2 to 10 mM) the DNA damage, expressed by the averaged median tail moment values, significantly increased in nuclei of all weeds studied. Using the Head Extent parameter of the Komet version 3.1, we have measured the diameter size of the nuclei of the 10 weed species either immediately after the isolation of the nuclei or after 20 or 45 min of treatment with alkaline buffer (pH > 13). According to the increase of the diameter of the nuclei (including the formed “halo”) resulting from the to alkaline buffer treatment, electrophoretic conditions (unwinding and electrophoresis time) for the Comet assay can be selected for the individual weed species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Angelis, K.J., Dušinská, M., Collins, A.R.: Single cell gel electrophoresis: Detection of DNA damage at different levels of sensitivity.-Electrophoresis 20: 2133–2138, 1999.

    Google Scholar 

  • Collins, A.R., Dobson, V.L., Dušinská, M., Kennedy, G., Štětina, R.: The comet assay: what can it really tell us?-Mutat. Res. 375: 183–193, 1997.

    Google Scholar 

  • Gichner, T., Badayev, S.A., Demchenko, S.I., Relichová, J., Sandhu, S.S., Usmanov, P.D., Usmanova, O., Velemínský, J.: Arabidopsis assay for mutagenicity.-Mutat. Res. 310: 249–256, 1994.

    Google Scholar 

  • Gichner, T., Plewa, M.J.: Induction of somatic DNA damage as measured by single cell gel electrophoresis and point mutation in leaves of tobacco plants.-Mutat. Res. 401: 143–152, 1998.

    Google Scholar 

  • Gichner, T., Ptáček, O., Stavreva, D.A., Plewa, M.J.: Comparison of DNA damage in plants as measured by single cell gel electrophoresis and somatic leaf mutations induced by monofunctional alkylating agents.-Environ. mol. Mutagen. 33: 279–286, 1999.

    Google Scholar 

  • Gichner, T., Menke, M., Stavreva, D.A., Schubert, I.: Maleic acid induces genotoxic effects but no DNA damage detectable by the Comet assay in tobacco and field beans.-Mutagenesis 15: 385–389, 2000a.

    Google Scholar 

  • Gichner, T., Ptáček, O., Stavreva, D.A., Wagner, E.D., Plewa, M.J.: A comparison of DNA repair using the Comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation.-Mutat. Res. 470: 1–9, 2000b.

    Google Scholar 

  • Koppen, G., Verschaeve, L.: The alkaline comet test on plant cells: A new genotoxicity test for DNA strand breaks in Vicia faba root cells.-Mutat. Res. 360: 193–200, 1996.

    Google Scholar 

  • Koppen, G., Angelis, K.J.: Repair of X-ray induced DNA damage measured by the Comet assay in roots of Vicia faba.-Environ. mol. Mutagen. 32: 281–285, 1998.

    Google Scholar 

  • Lovell, D.P., Tomas, G., Dulbow, R.: Issues related to the experimental design and subsequent statistical analysis of in vivo and in vitro comet studies.-Teratog. Carcinog. Mutagen. 19: 109–119, 1999.

    Google Scholar 

  • Menke, M., Chen, I.-P., Angelis, K.J., Schubert, I.: DNA damage and repair in Arabidopsis thaliana as measured by the comet assay after treatment with different classes of genotoxins.-Mutat. Res. 494: 87–94, 2001.

    Google Scholar 

  • Ptáček, O., Stavreva, D.A., Kim, J.K., Gichner, T.: Induction of DNA damage as measured by the Comet assay and the yield of somatic mutations in gamma-irradiated tobacco seedlings.-Mutat. Res. 491: 17–23, 2001.

    Google Scholar 

  • Tice, R.R., Agurell, E., Anderson, D., Burlinson, B., Hartmann, A., Kobayashi, H., Miyamae, Y., Rojas, E., Ryu, J.-C., Sasaki, Y.F.: Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing.-Environ. mol. Mutagen. 35: 206–221, 2000.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gichner, T., Mühlfeldová, Z. Induced DNA Damage Measured by the Comet Assay in 10 Weed Species. Biologia Plantarum 45, 509–516 (2002). https://doi.org/10.1023/A:1022360722422

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022360722422

Navigation