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Dynamic change of DNA methylation and cell redox state at different micropropagation phases in birch

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Phase change-related epigenetic and physiological changes in the micropropagation process of Betula platyphylla.

Abstract

To evaluate the epigenetic regulation and the cell redox state during micropropagation, Betula platyphylla was used to regenerate plants from callus. Axillary bud, callus (20 days after induction), aged callus (40 days after induction), budding callus, regenerating shoots and regenerated root were estimated for the cell redox state using 18 biochemical parameters, such as the steady-state levels of reactive oxygen, content of peroxidation products and enzymatic or non-enzymatic protective systems. These biochemical parameters at different developmental phases were found to be significantly different. DNA methylation and corresponding enzyme activities were also investigated at different phases. The average percentage of methylated CCGG sites in the three birch lines varied from 11.92 to 17.03 % at different phases. The level of DNA methylation in the axillary bud and young callus was 13.84 and 11.92 %, respectively. It increased to 12.36 % at budding callus phase and to 15.24 % in the regenerated bud. At rooting phase, DNA methylation was observed to be the highest (17.03 %). When compared between different phases, younger callus was found to have a lower level of DNA methylation (11.92 %) than that of the older callus (14.50 %), indicating that DNA methylation took place as the callus aged. The results also showed that CCGG site methylation patterns (hemi-methylation of external C sites and full methylation of internal C sites) were different at various differentiation phases. The results of principal component analysis clearly demonstrated that there were phase change-related epigenetic and physiological changes in the micropropagation process. These data lead to the identification of biochemical parameters during the process of micropropagation and demonstrate a dynamic connection between plant micropropagation in vitro, cell redox state and DNA methylation.

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Abbreviations

APX:

Ascorbate peroxidase

CAT:

Catalase

DHAR:

Dehydroascorbate reductase

GR:

Glutathione reductase

GST:

Glutathione S-transferase

mC:

Methylcytosine

MDHAR:

Monodehydroascorbate reductase

ROS:

Reactive oxygen species

SE:

Standard error

SOD:

Superoxide dismutase

POD:

Peroxidase

Lig:

Lignin

PAL:

Phenylalanineammonia-lyase

Chl a and b :

Chlorophylls a and b

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Author contribution statement

Conceived and designed the experiments: ZHAN YG and ZENG FS. Performed the experiments: ZENG FS, SUN FK and LUO W. Analyzed the data: ZENG FS and ZHAO XT. Contributed reagents/materials/analysis tools: ZHAN YG, ZENG FS and LIANG NS. Wrote the paper: ZENG FS and ZHAN YG.

Acknowledgments

This work was financially supported by the Fundamental Research Funds for the Central Universities (NO: 2572014DA04) and the National Natural Science Foundation of China (NO: J1210053, 31070531 and 31200463). We also thank the editor and two anonymous reviewers for many detailed and helpful comments that improved the quality of this manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University), Harbin, 150040, China

    Fan-Suo Zeng & Ya-Guang Zhan

  2. College of Life Science, Northeast Forestry University, Harbin, 150040, China

    Fan-Suo Zeng, Feng-Kun Sun, Nan-Song Liang, Xing-Tang Zhao, Wei Luo & Ya-Guang Zhan

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  1. Fan-Suo Zeng
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  2. Feng-Kun Sun
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Correspondence to Ya-Guang Zhan.

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Zeng, FS., Sun, FK., Liang, NS. et al. Dynamic change of DNA methylation and cell redox state at different micropropagation phases in birch. Trees 29, 917–930 (2015). https://doi.org/10.1007/s00468-015-1174-7

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