Assessment of epigenetic and phenotypic variation in Populus nigra regenerated via sequential regeneration
Open accessAbstract
Somatic variation has been demonstrated in tissue culture regenerated plants of many species. In the genus Populus , phenotypic variation caused by changes in 5-methylcytosine within the plant genome have been reported. To date, the phenotypic and epigenetic stability of plants regenerated from sequential regeneration has not been tested in trees. In this study, we detected DNA methylation of CCGG sites in regenerated plants of five generations in Populus nigra using methylation-sensitive amplified polymorphisms, and evaluated their growth performance and physiological traits. About 10.86–26.80% of CCGG sites in the regenerated plant genome were demethylated and 5.50–8.45% were methylated, resulting in significantly lower DNA methylation levels among all regenerated plants than among donor plants. We detected a significant difference in methylation levels between first regeneration regenerated plants (G1) and those of the other four generations (G2–G5); there were no significant differences among the four later generations. Therefore, the dramatic decrease in DNA methylation levels occurred only in the first and second poplar regenerations; levels then stabilized later in the regeneration process, indicating that two regeneration events were sufficient to change the methylation statuses of almost all CCGG sites sensitive to regeneration. Differences in growth and physiological traits were observed between regenerated plants and donor plants, but were significant only among plants of certain generations. Significant correlations were detected between methylation level and transpiration rate, net photosynthetic rate, peroxidase activity, and instant water utilization efficiency, indicating the involvement of epigenetic regulation in this unpredictable phenotypic variation.
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Epigenetic variability observed at the phenotype level reflects adaptations of cellular processes to constraints of in vitro cultivation, which usually implies sequential dedifferentiation and re-differentiation.
“This epigenetic variability, which frequently is observed at phenotype level, reflects, in reality, the adaptations of the cellular processes to the various constraints of the in vitro cultivation technology, that usually implies sequential dedifferentiation (formation of callus) and re-differentiation (regeneration of plants) [91,95,96,97,98,99,100].”
From Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants (2023) · cites this paper as [96]
The Introduction (page unavailable) links tissue culture to phenotypic variation and epigenetic changes, and the Discussion (page unavailable) reports methylation changes during regeneration. The paper does not establish that these changes are cellular adaptations to cultivation constraints or that in vitro cultivation usually involves sequential callus formation and plant regeneration; its Introduction instead describes somaclonal variation as thought to result from stress and disruption of normal cellular processes. (LLM, conf=0.82)
Experimental — accuracy not yet measuredIn Populus nigra, the methylation pattern decreased mainly in the first and second generations and became stable throughout subsequent generations.
“The conclusion of the study was that the methylation pattern decreased mainly in the first and second generations, and became stable throughout subsequent generations.”
From Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants (2023) · cites this paper as [96]
In Populus nigra regenerated from callus, all regenerated plants had significantly lower levels of DNA methylation, with considerable differences between the first generation and the other four generations, but no notable differences between G3, G4, and G5.
“The results indicated that all the regenerated plants had significantly lower levels of DNA methylation, with considerable differences between the first generation regenerated plants (G1) and those of the other four generations (G2–G5), but with no notable differences between the G3, G4, and G5 in vitro clones.”
From Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants (2023) · cites this paper as Zhang [96]
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- Authors
- Zhang, W., Wang, Y., Diao, S., Zhong, S., Wu, S., Wang, L., Su, X., & Zhang, B.
- Affiliations
- Chinese Academy of Forestry (CN)
- Jiangxi University of Water Resources and Electric Power (CN)
- Corresponding author
- Zhang, Bingyu
- Venue
- Frontiers in Plant Science
- Publisher
- Frontiers Media SA
- Published
- 2021-07-06
- Volume
- 12
- Pages
- 632088-632088
- ISSN
- 1664-462X (ISSN-L)
- PMC
- PMC8290414
- OpenAlex
- W3180895718
- Cited by
- 13
- References
- 36
- Citations by year
- 2026: 5 · 2025: 1 · 2024: 2 · 2023: 2 · 2022: 3
- FWCI
- 0.55
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Frontiers in Plant Science (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- National Natural Science Foundation of China (31770710, CAFYBB2020SZ002)
- Chinese Academy of Forestry (CAFYBB2020SZ002)
- State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
- Keywords
- Populus nigra, DNA methylation, epigenetic variation, somatic variation, tissue culture regeneration, methylation-sensitive amplified polymorphism, demethylation, transpiration rate, net photosynthetic rate, peroxidase activity, water use efficiency, growth performance, physiological traits
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