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Flow Cytometry, HPLC-RP, and metAFLP Analyses to Assess Genetic Variability in Somatic Embryo-Derived Plantlets of Gentiana pannonica Scop

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Abstract

Cytometric and molecular techniques were used to verify genetic uniformity among somatic embryo-derived plantlets of Gentiana pannonica Scop. Cytometric analysis of regenerants revealed absence of chromosomal changes and alterations in ploidy. However, reverse phase high pressure liquid chromatography detected higher levels of methylation in regenerated plants than those of control plants. These changes were further investigated using a quantitative molecular marker-based approach. This revealed that numerous tissue culture-induced variations, ∼3% (epi)mutations, were observed, including sequence variation and changes in methylation patterns. Moreover, complex patterns of variation, including combinations of genetic and epigenetic changes, were relatively high (ca. 9%). Overall, tissue culture-induced variation reached 16%; while, demethylation was lower than de novo methylation in heterozygotic material and similar in all regenerated plantlets.

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Acknowledgments

Authors express thanks to Dr. Kieth Harding (University of Abertay Dundee, UK) for his critical pre-review and suggestion which helped to improve the paper.

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Authors and Affiliations

  1. Botanical Garden–Center for Biological Diversity Conservation, Polish Academy of Sciences, Prawdziwka 2 Str., 02-973, Warsaw, Poland

    Agnieszka Fiuk & Jan J. Rybczyński

  2. Department of Plant Biochemistry and Physiology, Plant Breeding and Acclimatization Institute, Radzików, 05-870, Błonie, Poland

    Piotr T. Bednarek

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  1. Agnieszka Fiuk
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  2. Piotr T. Bednarek
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  3. Jan J. Rybczyński
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Correspondence to Agnieszka Fiuk.

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Fiuk, A., Bednarek, P.T. & Rybczyński, J.J. Flow Cytometry, HPLC-RP, and metAFLP Analyses to Assess Genetic Variability in Somatic Embryo-Derived Plantlets of Gentiana pannonica Scop. Plant Mol Biol Rep 28, 413–420 (2010). https://doi.org/10.1007/s11105-009-0167-3

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