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Alicja Tymoszuk,1 Dariusz Kulus,1 Alicja Kulpińska,1 Katarzyna Gościnna,2 Paulina Pietrzyk-Thel,3 Magdalena Osial3 1Laboratory of Horticulture and Landscape Architecture, Department of Biotechnology, Faculty of Agriculture and Biotechnology, Bydgoszcz University of Science and Technology, Bydgoszcz, Poland; 2Department of Agronomy and Food Processing, Faculty of Agriculture and Biotechnology, Bydgoszcz University of Science and Technology, Bydgoszcz, Poland; 3Department of Theory of Continuous Media and Nanostructures, Institute of Fundamental Technological Research, Polish Academy of Sciences, Warsaw, PolandCorrespondence: Alicja Tymoszuk, Laboratory of Horticulture and Landscape Architecture, Department of Biotechnology, Faculty of Agriculture and Biotechnology, Bydgoszcz University of Science and Technology, Bernardyńska 6, Bydgoszcz, 85-029, Poland, Email alicja.tymoszuk@pbs.edu.plPurpose: Chrysanthemum is one of the most popular ornamental plants worldwide. Its breeding remains a highly relevant topic. Nanotechnology significantly and interdisciplinarily contributes to the progress in modern horticulture. To date, there are no studies on the use of the proposed heavy metal-based nanoparticles in mutation breeding of ornamental plants.Methods: CdS NPs, Co3O4 NPs, and Fe3O4@Co NPs were synthesized and applied at a concentration of 75 mg·L− 1 in the in vitro internode culture of Chrysanthemum × morifolium (Ramat). Hemsl. ‘Lilac Wonder’.Results: The highest number of adventitious shoots was regenerated on the control and Fe3O4@Co NP-treated internodes, whereas the use of CdS NPs and Co3O4 NPs hampered regeneration. The NP-treated shoots, compared to the control, accumulated less flavonols and more anthocyanins and polyphenols, and exhibited increased antioxidant capacity. The highest activity of oxidative stress enzymes and the lowest chlorophyll content were noted in CdS NP-treated shoots. The tested nanoparticles also affected the further growth and development of plants during ex vitro greenhouse cultivation. The longest stems were found in Fe3O4@Co NP-treated plants, contrary to CdS NPs and Co3O4 NPs. The CdS NP-treated plants developed leaves with the smallest surface area, perimeter, length, and width. Evaluation of inflorescences revealed quantitative changes in anthocyanins content. The highest pigment content was found in ligulate flowers of Fe3O4@Co NP-treated plants. One individual with variegated leaves was phenotypically identified within Co3O4 NP-treated plants. Genetic variation was detected in 7– 8.1% of the plants studied. The SCoT marker system generated more bands and polymorphisms than RAPD. PCoA analysis revealed distinct genetic groupings, with the most altered genotype (treated with CdS NPs) classified as polymorphic by both marker systems. The other 11 polymorphic genotypes did not overlap between RAPD and SCoT analyses.Conclusion: Our results proved that nanoparticles can serve as a novel and valuable tool for plant breeding.Keywords: breeding, Chrysanthemum × morifolium, genetic marker, nanobiotechnology, phenotype, physiology |