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Genetic Variation Induced by Tissue and Organ Culture in Gentiana Species

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The Gentianaceae - Volume 2: Biotechnology and Applications

Abstract

This chapter presents the results of cytogenetic and molecular genetic studies on tissue and organ cultures from some Gentiana species. Cytogenetic analysis of calli from seven gentians showed that all of them (except for G. punctata ) consisted mainly of the cells with diploid and/or near-diploid chromosome sets with low anaphase aberration. Species specificity of cytogenetic structure was established for cell populations in vitro. An increase in the proportion of polyploid cells was found for G. acaulis callus after long-term culture. RAPD analysis of tissue cultures from six gentians showed that culture resulted in genetic changes, with the species differing in the level of genetic variation. RAPD and ISSR analyses of G. pneumonanthe and G. lutea genetic variation in tissue and organ cultures and regenerated plants revealed the most significant changes in isolated root cultures as well as in long-term cultured tissues. At the same time, direct and indirect regenerants had the least genetic difference compared to donor plant . The restriction maps of nuclear 18S-25S ribosomal DNA from G. acaulis , G.  punctata , G. lutea, and G. asclepiadea were constructed based on the results of blot hybridization. The decrease of 18S-25S rDNA amount was found in cultured tissues. In contrast to other species, G. lutea showed intragenome heterogeneity of rRNA genes as well as qualitative rDNA changes in tissue culture, such as the appearance of repeats with altered length. A relationship has been suggested between the peculiarities of the structural organization of the 18S-25S rRNA gene cluster and its rearrangements. Rearrangements of 5S rDNA were not found in cultures of Gentiana L. species, except for the variation in nucleotide methylation pattern at restriction sites.

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Acknowledgments

This research was supported by the National Academy of Sciences of Ukraine through the Targeted interdisciplinary programme of scientific research “Fundamentals of molecular and cellular biotechnology” and a research grant support from the State Fund for Fundamental Research of Ukraine (No 05.07/00219).

We are grateful to Dr. Olena G. Alkhimova (Institute of Molecular Biology and Genetics National Academy of Sciences of Ukraine) for kindly providing plasmids with 18S-25S and 5S rDNA sequences as well as valuable suggestions for preparation of the section of the manuscript concerning rDNA.

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Authors and Affiliations

  1. Institute of Molecular Biology and Genetics National Academy of Sciences of Ukraine, Akad. Zabolotnogo Str., 150, Kyiv, 03680, Ukraine

    Viktor A. Kunakh, Vitaliy M. Mel’nyk, Igor O. Andreev, Kateryna V. Spiridonova, Maryana O. Twardovska, Iryna I. Konvalyuk & Volodymyr I. Adonin

  2. Volodymyr Hnatiuk Ternopil National Pedagogical University, M. Kryvonis Str., 2, Ternopil, 46027, Ukraine

    Nadia M. Drobyk

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  1. Viktor A. Kunakh
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  2. Vitaliy M. Mel’nyk
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  4. Igor O. Andreev
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  8. Volodymyr I. Adonin
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Correspondence to Viktor A. Kunakh .

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Editors and Affiliations

  1. Botanical Garden-Center for Biological Diversity Conservation, Polish Academy of Sciences, Warsaw, Poland

    Jan J. Rybczyński

  2. Plant and Crop Sciences Division, University of Nottingham School of Biosciences, Loughborough, United Kingdom

    Michael R. Davey

  3. Botanical Garden-Center for Biological Diversity Conservation, Polish Academy of Sciences, Warsaw, Poland

    Anna Mikuła

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Kunakh, V.A. et al. (2015). Genetic Variation Induced by Tissue and Organ Culture in Gentiana Species. In: Rybczyński, J., Davey, M., Mikuła, A. (eds) The Gentianaceae - Volume 2: Biotechnology and Applications. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-54102-5_9

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