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Classical mutation breeding using physical factors is a common breeding method for ornamental crops. The aim of our study was to examine the utility of ovaries excised from irradiated inflorescences of Chrysanthemum × morifolium (Ramat.) as explants for breeding purposes. We studied the in vitro regeneration capacity of the ovaries of two chrysanthemum cultivars: ‘Profesor Jerzy’ and ‘Karolina’ preceded by irradiation with high-energy photons (total dose 5, 10 and 15 Gy) and high-energy electrons (total dose 10 Gy). Growth and inflorescence parameters of greenhouse acclimatized regenerants were recorded, and ploidy level was estimated with flow cytometry. The strong impact of genotype on regeneration efficiency was recorded—cultivar ‘Karolina’ produced only 7 viable shoots, while ‘Profesor Jerzy’ produced totally 428 shoots. With an increase of irradiation dose, the regeneration decreased, the least responsive were explants irradiated with 15 Gy high-energy photons and 10 Gy high-energy electrons. Regenerants of ‘Profesor Jerzy’ obtained from these explants possessed shorter stem and flowered later. The highest number of stable, color and shape inflorescence variations were obtained from explants treated with 10 Gy high-energy photons. Variations of inflorescences were predominantly changes of shape—from full to semi-full. New color phenotypes were dark yellow, light yellow and pinkish, among them only the dark yellow phenotype remained stable during second year cultivation. None of the regenerants were haploid. The application of ovaries irradiated within the whole inflorescence of chrysanthemum can be successfully applied in the breeding programs, provided the mother cultivar regenerate in vitro efficiently.

Ovaries of Chrysanthemum Irradiated with High-Energy Photons and High-Energy Electrons Can Regenerate Plants with Novel Traits