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Variations in leaf morphology and DNA methylation following in vitro culture of Malus xiaojinensis

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Abstract

Juvenile trees of Malus xiaojinensis produce leaves with three to five lobes; however, adult trees do not. Thus, M. xiaojinensis serves as a model plant for studies on rejuvenation of woody perennials. To better understand effects of in vitro culture, number of passages, and plant growth regulators on observed somaclonal variations in leaf morphology, total levels of DNA methylation and expression of associated genes, including DRM2 and MET1, of apical shoot meristems of M. xiaojinensis, were evaluated. Following 17 passages of nodal stem segment culture and four passages of callus culture, levels of DNA methylation and expression of DRM2 and MET1 were determined, and found to decrease from the 11th passage of nodal stem segment culture or adventitious bud culture and from the first passage of callus culture. Levels of DNA methylation increased with increasing 2,4-dichlorophenoxyacetic acid concentrations, but decreased significantly with increasing 6-benzyladenine (BA) concentrations. Expression of DRM2 and MET1 in regenerated plantlets were negatively correlated with BA concentrations in the culture medium. Overall, less than 11 passages of nodal stem segment or adventitious bud culture did not contribute to incidence of somaclonal variation, even in the presence of low concentrations of BA (<0.5 mg/L) in the culture medium. Moreover, when nodal stem segment cultures were maintained for up to 17 passages, complete rejuvenation was observed. However, epigenetic changes were detected following the first passage of callus cultures.

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Acknowledgments

This research was partially funded by the National Special Funds for Scientific Research on Public Causes (Agriculture) (201203075), the Modern Agricultural Industry Technology System (Apple)(CARS-28) and the Beijing Natural Science Foundation(6071002), China.

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  1. Institute for Horticultural Plants, China Agricultural University, Beijing, 100193, People’s Republic of China

    Huang Huang, Sha Sha Han, Yi Wang, Xin Zhong Zhang & Zhen Hai Han

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  1. Huang Huang
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  2. Sha Sha Han
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  3. Yi Wang
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  4. Xin Zhong Zhang
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  5. Zhen Hai Han
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Correspondence to Zhen Hai Han.

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Huang Huang and Sha Sha Han contributed equally to this work.

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Huang, H., Han, S.S., Wang, Y. et al. Variations in leaf morphology and DNA methylation following in vitro culture of Malus xiaojinensis . Plant Cell Tiss Organ Cult 111, 153–161 (2012). https://doi.org/10.1007/s11240-012-0179-9

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  • DOI: https://doi.org/10.1007/s11240-012-0179-9

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