DNA methylation changes and TE activity induced in tissue cultures of barley (Hordeum vulgare L.)
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BACKGROUND: In vitro plant regeneration via androgenesis or somatic embryogenesis is capable of inducing (epi)mutations that may affect sexual progenies. While epimutations are associated with DNA methylation, mutations could be due to the movement of transposons. The common notion is that both processes are linked. It is being assumed that demethylation activates transposable elements (TEs). Analysis of methylation changes and their relation with TEs activation in tissue cultures requires uniquely derived donor plants (Ds), their regenerants (Rs) and respective progeny (Ps) that would allow discrimination of processes not related to changes introduced via in vitro cultures. Moreover, a set of methods (RP-HPLC, SSAP, and MSTD) is needed to study whether different TEs families are being activated during in vitro tissue culture plant regeneration and whether their activity could be linked to DNA methylation changes or alternative explanations should be considered. RESULTS: The in vitro tissue culture plant regeneration in barley was responsible for the induction of DNA methylation in regenerants and conservation of the methylation level in the progeny as shown by the RP-HPLC approach. No difference between andro- and embryo-derived Rs and Ps was observed. The SSAP and MSTD approach revealed that Ds and Rs were more polymorphic than Ps. Moreover, Rs individuals exhibited more polymorphisms with the MSTD than SSAP approach. The differences between Ds, Rs and Ps were also evaluated via ANOVA and AMOVA. CONCLUSIONS: Stressful conditions during plant regeneration via in vitro tissue cultures affect regenerants and their sexual progeny leading to an increase in global DNA methylation of Rs and Ps compared to Ds in barley. The increased methylation level noted among regenerants remains unchanged in the Ps as indicated via RP-HPLC data. Marker-based experiments suggest that TEs are activated via in vitro tissue cultures and that, independently of the increased methylation, their activity in Rs is greater than in Ps. Thus, the increased methylation level may not correspond to the stabilization of TEs movement at least at the level of regenerants. The presence of TEs variation among Ds that were genetically and epigenetically uniform may suggest that at least some mobile elements may be active, and they may mask variation related to tissue cultures. Thus, tissue cultures may activate some TEs whereas the others remain intact, or their level of movement is changed. Finally, we suggest that sexual reproduction may be responsible for the stabilization of TEs.
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In Hordeum vulgare micropropagation, no variation was observed.
“No variation observed”
From Somaclonal Variation and Clonal Fidelity in Commercial Micropropagation: Challenges and Perspectives (2025) · cites this paper as [159]
The Results report no apparent morphological changes in barley regenerants or their progeny relative to donor plants. But the RP-HPLC Results report increased DNA methylation in regenerants, so the unqualified claim that no variation was observed is too broad. Our copy provides section labels but no page numbers for these passages. (LLM, conf=0.88)
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- Authors
- Orłowska, R., Machczyńska, J., Oleszczuk, S., Zimny, J., & Bednarek, P. T.
- Affiliations
- Plant Breeding and Acclimatization Institute - National Research Institute (PL)
- Corresponding author
- Bednarek, Piotr Tomasz
- Venue
- Journal of Biological Research-Thessaloniki
- Published
- 2016-08-08
- Volume
- 23
- Issue
- 1
- Pages
- 19-19
- ISSN
- 1790-045X (ISSN-L)
- 2241-5793
- PMC
- PMC4977862
- OpenAlex
- W2511231834
- Cited by
- 53
- References
- 80
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 6 · 2023: 2 · 2022: 10 · 2021: 11 · 2020: 7 · 2019: 6 · 2018: 4 · 2017: 2
- FWCI
- 2.08
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Journal of Biological Research - Thessaloniki (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Genetic and Mutation Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- DNA methylation, transposable elements, transposon activation, barley, in vitro plant regeneration, androgenesis, somatic embryogenesis, epimutations, epigenetic inheritance, transposon polymorphism, methylation-specific PCR
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