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Morphological, physiological and genomic comparisons between diploids and induced tetraploids in Anemone sylvestris L.

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Abstract

The objective of this research was to induce mitotic chromosome doubling in Anemone sylvestris L. The mitosis inhibitor oryzalin was directly added to the induction medium at 1, 2, 5, 10 and 15 μM for 8, 10 or 12 weeks of cultivation. Three tetraploid plants (2n = 4x = 32), 0.8% (polyploidization efficiency), were obtained from diploid plants (2n = 2x = 16) in three treatments (1 μM for 10 weeks, 5 μM for 8 weeks and 8 μM for 10 weeks). Ploidy level was confirmed by flow cytometry. Morphological characteristics (e.g. flower diameter, total plant height, leaf area) and chlorophyll content differences between diploid and tetraploid A. sylvestris were observed together with polyphenol content and antioxidant activity. The inter primer binding sites markers were used for evaluation of polymorphism. New genotypes with different morphological and biological characteristics were obtained through somatic polyploidization. The tetraploid plants were stronger, more vigorous and had an early flowering, which is essential for its use as an ornamental plant. The iPBS analysis showed unique amplicons that can be used for the purposes of molecular identification of tetraploid plants of A. sylvestris in the future. The results demonstrate the first report of in vitro induction of tetraploids of A. sylvestris.

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Acknowledgements

This study was supported by Project Nos. 20175009 and 20115003 of University Internal Grant Agency, Czech University of Life Sciences Prague, by European Community under Project Nos. 26220220180 Building Research Centre “AgroBioTech”, by Project Nos. 17-25536Y of the Czech Science Foundation and by Silva Tarouca Research Institute for Landscape and Ornamental Gardening (the institutional support VUKOZ-IP-00027073). The authors thank to prof. Jaroslav Doležel and Jana Čížková, Ph.D. from Institute of Experimental Botany, The Czech Academy of Sciences, the Czech Republic, for their help in the flow-cytometric analyses.

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Authors and Affiliations

  1. Faculty of Tropical AgriSciences, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21, Suchdol, Czech Republic

    Pavla Zahumenická & Eloy Fernández

  2. Department of Phytoenergy and Biodiversity, The Silva Tarouca Research Institute for Landscape and Ornamental Gardening, Květnové nám. 391, 252 43, Průhonice, Czech Republic

    Jana Šedivá

  3. Faculty of Agrobiology and Food Resources, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76, Nitra, Slovakia

    Jana Žiarovská

  4. Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21, Suchdol, Czech Republic

    José Luis Ros-Santaella

  5. Faculty of Environmental Sciences, Czech University of Life Sciences Prague, Kamýcká 129, Prague 6, 165 21, Suchdol, Czech Republic

    Domingo Martínez-Fernández

  6. Department of Science, Basilicata University, V.le Ateneo Lucano 10, 85100, Potenza, Italy

    Daniela Russo & Luigi Milella

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  1. Pavla Zahumenická
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  2. Eloy Fernández
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  3. Jana Šedivá
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  4. Jana Žiarovská
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  5. José Luis Ros-Santaella
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  8. Luigi Milella
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Contributions

PZ and EF established in vitro culture, realized induced polyploidization, flow cytometry analyses and morphological observations in vivo. JŠ transferred plants in ex vitro conditions and realized morphological analyses. JŽ was responsible for iPBS analyses. JLRS and DMF assessed chlorophyll content and made statistical analyses. DR and LM assessed chemical analysis and antioxidant activity. EF and LM designed the workflow of the experiments.

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Correspondence to Eloy Fernández.

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Communicated by Sergio J. Ochatt.

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Zahumenická, P., Fernández, E., Šedivá, J. et al. Morphological, physiological and genomic comparisons between diploids and induced tetraploids in Anemone sylvestris L.. Plant Cell Tiss Organ Cult 132, 317–327 (2018). https://doi.org/10.1007/s11240-017-1331-3

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