Various plant species can be selected for environmental testing, including pearl millet (Pennisetum glaucum (L.) R. Br), a globally significant cereal crop. This study aims to assess millet's suitability as a species for ecotoxicological tests, examining (1) germination and initial development dynamics, (2) the minimum seed quantity for reliable sampling, (3) optimal experimental design with replication numbers, (4) suitability of positive control, and (5) the effectiveness of the protocol in evaluating toxic effects of environmental pollutants. Millet exhibited rapid and uniform germination as well as consistent initial seedling development. To establish the minimum number of seeds required for reliable experimentation, germination, and seedling growth were compared across plots containing 10, 25, and 50 seeds. Consequently, 10 seeds per plot were chosen for subsequent experiments to reduce labor and costs while maintaining reliability. To validate the selected experimental design, and to establish a positive control for assays, aluminum was used as a toxic element at concentrations ranging from 10-2 to 10-6 M. While aluminum did not affect the final percentage of germinated seeds, it did exhibit an impact on the Germination Speed Index (GSI). Significant differences in root and aerial growth, and with fresh weight, were observed. The 10-3M concentration was chosen as the positive control as the 10-2 concentration showed extreme toxicity. To assess the applicability of the established protocol in determining the toxic effects of environmental pollutants, millet roots were exposed to the toxic agents atrazine, cadmium, methyl methane sulfonate (MMS), and Spent pot liner (SPL). Millet demonstrated sensitivity and efficiency in response to these tests. In conclusion, millet proves to be an effective species for the toxicological risk assessment of environmental pollutants.
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