Open dumping of fly ash in fly ash basins has significant adverse environmental impacts due to its elevated trace element content. In situ biomonitoring of genotoxicity is of practical value in realistic hazard identification of fly ash. Genotoxicity of openly disposed fly ash to natural plant populations inhabiting fly ash basins has not been investigated. DNA damage, and concentrations of As, Co, Cr, Cu and Ni in the leaves of natural populations of Cassia occidentalis growing at two contrasting sites-one having weathered fly ash (fly ash basin) and the other having soil (reference site) as plant growth substrates-were assessed. The foliar concentrations of As, Ni and Cr were two to eight fold higher in plants growing on fly ash as compared to the plants growing on soil, whereas foliar concentrations of Cu and Co were similar. We report, for the first time, based upon comet assay results, higher levels of DNA damage in leaf tissues of Cassia occidentalis growing wild on fly ash basin compared to C. occidentalis growing on soil. Correlation analysis between foliar DNA damage and foliar concentrations of trace elements suggests that DNA damage may perhaps be associated with foliar concentrations of As and Ni. Our observations suggest that (1) fly ash triggers genotoxic responses in plants growing naturally on fly ash basins; and (2) plant comet assay is useful for in situ biomonitoring of genotoxicity of fly ash.