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

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• 作者：Huang Huang (1)
  Sha Sha Han (1)
  Yi Wang (1)
  Xin Zhong Zhang (1)
  Zhen Hai Han (1) rschan@cau.edu.cn
• 关键词：DNA methylation &#8211 ; Malus xiaojinensis &#8211 ; Micro ; propagation &#8211 ; Somaclonal variation &#8211 ; Tissue culture
• 刊名：Plant Cell, Tissue and Organ Culture
• 出版年：2012
• 出版时间：November 2012
• 年：2012
• 卷：111
• 期：2
• 页码：153-161
• 全文大小：508.4 KB
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• 作者单位：1. Institute for Horticultural Plants, China Agricultural University, Beijing, 100193 People鈥檚 Republic of China
• ISSN：1573-5044

文摘

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.