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Gladiolus is one of the most important lucrative cut flower crops that is commercially cultivated worldwide due to its various spike forms, size, and shape and color combinations. In order to further increase the commercial and horticultural value by improving the ornamental traits of gladiolus ‘White Prosperity’, polyploidy was induced by soaking gladiolus corms in different colchicine concentrations (0.1%, 0.2% and 0.3%) for 24 h. Different colchicine concentrations had a little effect on sprouting and survival percentage but it significantly delayed the emergence of sprouts. About one third decreases in plant height along with reduction in number of leaves per plant, leaf area, length and width, chlorophyll content, diameter and number of cormlets per corm was observed in treated plants. Colchicine at 0.1% concentration improved the ornamental value of gladiolus by increasing vase life whereas colchicine at 0.3% was effective in increasing floret diameter. However, the colchicine treated plants exhibited delayed and reduced percentage of flowering corms. Pollen and stomatal study was done for the identification of polyploidy and it showed that both pollen and stomata size were increased while stomatal density and pollen fertility was significantly reduced in polyploid plants. Induction of polyploidy (mixoploids + octoploids) was achieved in all concentrations, however 0.2% and 0.3% concentrations of colchicine were effective for producing large number of polyploid plants. But at 0.1% concentration of colchicine, majority of plants did not show any change in their original ploidy level (tetraploid). These putative polyploids may be helpful for further improvement in ornamental and horticultural value of gladiolus.