Evaluation of the Alkaline Comet Assay Conducted With the Wetlands Plant Bacopa monnieri L. as a Model for Ecogenotoxicity Assessment

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Wetlands play a key role in maintaining environmental quality, and wetlands plants could serve as model organisms for determining the genotoxic effects of pollutants contaminating these areas. In the present study, DNA damage was evaluated in a wetlands plant, Bacopa monnieri L., as a potential tool for the assessment of ecogenotoxicity. The Comet assay was used for detecting DNA damage in B. monnieri exposed to two model mutagens, ethyl methanesulfonate (EMS) and methyl methanesulfonate (MMS). Significant (P < 0.05) dose-dependent increases in DNA damage were observed following treatments conducted by exposing both isolated nuclei (acellular or in vitro exposure) and whole plants (in vivo exposure) to 0.01-5 mM EMS and 0.05-100 μM MMS for 2 hr at (26 ± 2)°C. The assay was then used to evaluate the genotoxic potential of cadmium (Cd), a wetlands contaminant. In vitro exposure of nuclei from untreated leaves to 0.001-200 μM Cd for 2 hr resulted in significant (P < 0.05) levels of DNA damage. Cd concentrations ≥0.01 μM induced DNA damage as evidenced by increases in the Olive tail moment. In vivo exposure of plants to 0.01-500 μM Cd for 2, 4, and 18 hr resulted in dose- and time-dependent increases in DNA damage in the nuclei isolated from roots and leaves. Cd-induced DNA damage was greater in roots than leaves. To our knowledge, this is the first report describing the use of a wetlands plant for genotoxicity assessment, using the Comet assay.

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