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Colchicine is one of the important aqueous solutions that has been used to induce mutation or ploidy in many plant species long ago. However, its proper concentrations plays a vital role, increased or inappropriate concentrations may cause mortality, stunted growth, morphological deformation, etc. in plant species. Thus, this study was conducted to evaluate the different colchicine concentrations on the growth and development of gladiolus corms. Gladiolus corms were treated with 0.2%, 0.4%, and 0.6% concentrations for 24 hours. Colchicine toxicity was evaluated during the early growth stage showed a higher concentration of 0.6% significantly reduced corm survival (47%) and caused a 51.9% survival reduction over control. In terms of growth parameters, all concentrations of colchicine reduced plant height with a number of leaves along with inducing different forms of morphological abnormalities and chlorophyll mutants. To optimize the dose for successful ploidy induction, LD30 and LD50 on mortality rate through an analysis were calculated to be 0.31% and 0.57% whereas GR50 based on plant height and the number of leaves reduction was found to be 0.17% and 0.34%. Hence, the results from this study could be used in the future for further breeding programs by reducing the genotoxicity of colchicine on gladiolus.