Does somaclonal variation play advantageous role in conservation practice of endangered species?: comprehensive genetic studies of in vitro propagated plantlets of Viola stagnina Kit. (Violaceae)

Piotr Żabicki , Elwira Śliwińska , Józef Mitka , Agnieszka Sutkowska , Monika Tuleja , Grzegorz Migdałek , Justyna Żabicka , Aneta Słomka , Monika Kwiatkowska , Elżbieta Kuta


In vitro regeneration of Viola stagnina Kit., endangered in most part of its European distribution range, was successfully obtained based on the newly developed protocol. Adventitious shoots via direct and indirect organogenesis were induced on leaf blade and petiole fragments on solidified Murashige and Skoog (MS) medium supplemented with 0.5 or 1 mg l−1 thidiazuron, respectively. Shoots were rooted on half-strength MS medium with 2% sucrose and 0.5 mg l−1 indole-3-acetic acid (IAA), and plantlets were successfully acclimatized. Sixty-five of the regenerated plants (72% of isolated shoots cultured on rooting medium) survived at the experimental plot conditions. Among recovered via organogenesis plants, individuals ‘true-to-type’ derived from initial plant G1 and also plants genetically distant from initial plants were detected by ISSR markers. In all groups of clones genetic indices (number of genotypes, polymorphic markers, gene diversity, total gene diversity, mean gene diversity) were lower than in natural populations. Regenerated plantlets had the same genome size estimated by flow cytometry as initial material and plants from natural populations. They developed chasmogamous flowers with highly viable pollen grains (over 90%), cleistogamous flowers, and set seeds from both flower types in the first and second seasons cultivated at experimental plots. This is the first report of a successfully developed micropropagation protocol of V. stagnina, and the first detailed genetic analysis of recovered plants with the use of ISSR markers and genome size measurements allowing to discuss the advantageous role of somaclonal variation in ex situ plant conservation with the use of in vitro micropropagation.
Author Piotr Żabicki
Piotr Żabicki,,
, Elwira Śliwińska
Elwira Śliwińska,,
, Józef Mitka
Józef Mitka,,
, Agnieszka Sutkowska (FoAE / DoPBaS)
Agnieszka Sutkowska,,
- Department of Plant Breeding and Seed Science
, Monika Tuleja
Monika Tuleja,,
, Grzegorz Migdałek
Grzegorz Migdałek,,
, Justyna Żabicka
Justyna Żabicka,,
, Aneta Słomka
Aneta Słomka,,
, Monika Kwiatkowska
Monika Kwiatkowska,,
, Elżbieta Kuta
Elżbieta Kuta,,
Journal seriesPlant Cell Tissue and Organ Culture, [Plant Cell, Tissue and Organ Culture], ISSN 0167-6857, e-ISSN 1573-5044, (N/A 100 pkt)
Issue year2019
Publication size in sheets0.65
Keywords in EnglishIn vitro organogenesis, Plant regeneration, Genome size, Somaclonal variation, ISSR markers, Viola
ASJC Classification1108 Horticulture
Internal identifierWRE/2019/31
Languageen angielski
Score (nominal)100
Score sourcejournalList
Publication indicators WoS Citations = 1; Scopus SNIP (Source Normalised Impact per Paper): 2017 = 1.122; WoS Impact Factor: 2018 = 2.2 (2) - 2018=2.232 (5)
Citation count*
Additional fields
FinansowanieThis work was funded in part by the Dean of the Faculty of Biology and Earth Sciences of Jagiellonian University (Project DS/MND/WBiNoZ/IB/25/2012 for P. Żabicki).
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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