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Analysis of ovary DNA methylation during delayed fertilization in hazel using the methylation-sensitive amplification technique

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Abstract

Key message

Preferential gene-body methylation is a key feature of the mechanism that initiates ovary and ovule development after pollination in hazel.

Abstract

Hazel (Corylus avellana L.) produces edible nuts of great economic value and has unique delayed fertilization characteristics. The ovary in hazel starts to develop only after pollination, and a growth signal from pollen tubes is thought to be the key factor that triggers this process. DNA methylation may play an important role in the epigenetic activation of ovary development in hazel; however, changes in DNA methylation during the delayed fertilization process remain unknown. In this study, we assessed DNA methylation changes during pre- and post-pollination stages using the methylation-sensitive amplification polymorphism (MSAP) technique. A total of 760 5′-CCGG-3′ amplified sites were detected by MSAP assay. The total methylation ratio of hazel was relatively high, ranging from 44.61 to 48.68 %, while after pollination, it decreased slightly by approximately 4 %. MSAP bands were significantly different between pre- and post-pollination stages, and a total of 15 differentially amplified bands (L01–L15) were cloned for sequence analysis. Most of the fragments were homologous to gene exons or mitochondrial sequences. Quantitative reverse transcription polymerase chain reaction showed that the gene-body methylation of three differentially amplified bands (L08, L10, and L13) was closely associated with their gene expression level. Bisulfite sequencing of L08 showed that cytosine methylation and demethylation changes varied in different pre- and post-pollination stages. In conclusion, epigenetic changes probably represent an important mechanism that initiates ovary and ovule development after pollination in hazel in which preferential gene-body methylation is a key feature.

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Acknowledgments

This work was funded by the National Natural Science Foundation of China (31370683 and 31070610) and the Science and Technology Project of Jilin Province (20130102052JC and 20140101145JC).

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Authors and Affiliations

  1. College of Life Sciences, Jilin Normal University, Siping, 136000, Jilin Province, China

    Yunqing Cheng, Ju Wang, Jianfeng Liu, Yixin Zhao, Wanting Geng & Huidi Zhang

Authors
  1. Yunqing Cheng
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  2. Ju Wang
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  3. Jianfeng Liu
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  4. Yixin Zhao
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  5. Wanting Geng
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  6. Huidi Zhang
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Corresponding author

Correspondence to Jianfeng Liu.

Additional information

Communicated by T. Moriguchi.

Electronic supplementary material

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Supplementary material 1

Fig. S1. Methylation-sensitive amplification polymorphism (MSAP) analysis of DNA methylation changes during pre- and post-pollination in hazel (Part 1) (jpg 21 kb)

Supplementary material 2

Fig. S2. Methylation-sensitive amplification polymorphism (MSAP) analysis of DNA methylation changes during pre- and post-pollination in hazel (Part 2) (jpg 19 kb)

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Fig. S3. Methylation-sensitive amplification polymorphism (MSAP) analysis of DNA methylation changes during pre- and post-pollination in hazel (Part 3) (jpg 20 kb)

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Figs. S4–18. Sequencing data of differentially amplified bands (L01–L15) derived from methylation-sensitive amplification polymorphism (MSAP) assay (seq 1 kb)

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Cheng, Y., Wang, J., Liu, J. et al. Analysis of ovary DNA methylation during delayed fertilization in hazel using the methylation-sensitive amplification technique. Acta Physiol Plant 37, 231 (2015). https://doi.org/10.1007/s11738-015-1984-7

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  • DOI: https://doi.org/10.1007/s11738-015-1984-7

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