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Analysis of DNA methylation of perennial ryegrass under drought using the methylation-sensitive amplification polymorphism (MSAP) technique

Molecular Genetics and Genomics volume 289, pages 1075–1084 (2014)Cite this article

Abstract

Perennial ryegrass (Lolium perenne), an excellent grass for forage and turf, is widespread in temperate regions. Drought is an important factor that limits its growth, distribution, and yield. DNA methylation affects gene expression and plays an important role in adaptation to adverse environments. In this study, the DNA methylation changes in perennial ryegrass under drought stress were assessed using methylation-sensitive amplified polymorphism (MSAP). After 15 days of drought stress treatment, the plant height was less than half of the control, and the leaves were smaller and darker. Genome-wide, a total of 652 CCGG sites were detected by MSAP. The total methylation level was 57.67 and 47.39 % in the control and drought treatment, respectively, indicating a decrease of 10.28 % due to drought exposure. Fifteen differentially displayed DNA fragments in MSAP profiles were cloned for sequencing analysis. The results showed that most of the genes involved in stress responses. The relative expression levels revealed that three demethylated fragments were up-regulated. The expression of a predicted retrotransposon increased significantly, changing from hypermethylation to non-methylation. Although the extent of methylation in two other genes decreased, the sites of methylation remained, and the expression increased only slightly. All of these results suggested that drought stress decreased the total DNA methylation level in perennial ryegrass and demethylation up-regulated related gene expressions and that the extent of methylation was negatively correlated with expression. Overall, the induced epigenetic changes in genome probably are an important regulatory mechanism for acclimating perennial ryegrass to drought and possibly other environmental stresses.

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Acknowledgments

We thank the anonymous reviewers for their constructive comments and suggestions for improving the manuscript. This research was supported by the National High Technology Research and Development Program of China (863 Program) (2009AA10Z108, 2008AA10Z409) and the West Light Foundation of The Chinese Academy of Sciences (Y3C4011100).

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Affiliations

  1. Chengdu Institute of Biology, Chinese Academy of Sciences, No 9, Section 4, Renmin South Road, Chengdu, 610041, China

    Xiao-Mei Tang, Xiang Tao, Yan Wang, Dan Li, Hong Yang & Xin-Rong Ma

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Xiao-Mei Tang

  3. Department of Information Engineering, Jilin Business and Technology College, Changchun, 130062, China

    Dong-Wei Ma

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  1. Xiao-Mei Tang
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  2. Xiang Tao
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  3. Yan Wang
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Correspondence to Xin-Rong Ma.

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Communicated by S. Hohmann.

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Tang, XM., Tao, X., Wang, Y. et al. Analysis of DNA methylation of perennial ryegrass under drought using the methylation-sensitive amplification polymorphism (MSAP) technique. Mol Genet Genomics 289, 1075–1084 (2014). https://doi.org/10.1007/s00438-014-0869-6

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  • DOI: https://doi.org/10.1007/s00438-014-0869-6

Keywords

  • Perennial ryegrass
  • MSAP
  • DNA methylation/demethylation
  • Drought
  • Gene expression