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DNA methylation analysis of sterile and fertile CMS-C hybrids and their parents in maize

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Abstract

C-type cytoplasmic male sterile (CMS-C) line plays an important role in hybrid seed production in maize. However, mechanisms of pollen abortion and fertility restoration remain unclear. This study aimed to investigate the mechanisms of CMS-C pollen abortion and fertility restoration, particularly based on epigenetics, and to understand the relationship between male fertility performance and DNA methylation status. Methylation-sensitive amplification polymorphism (MSAP) technique was conducted to analyze DNA methylation levels and patterns of tassels at pollen-mother cell, tetrad, mononuclear, and binuclear stages among four half-sib hybrids and their parents with different fertility. Results showed that DNA methylation levels in fertility restored hybrids were higher than those in sterility maintained hybrids. Nine CCGG sites, which displayed methylation polymorphism between male-fertile and male-sterile hybrids, were screened. To validate MSAP results, we performed methylation-sensitive PCR (MS-PCR) and found consistent observations in cloned methylation sites. Interestingly, a specific site named 16–1 was discovered in the region of Rf5 gene, which is one of the restorer genes of CMS-C. Thus, DNA methylation may participate in regulating the expression of fertility restorer genes of maize CMS-C.

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Abbreviations

CMS:

Cytoplasmic male sterile

CMS-C:

C-type cytoplasmic male sterile

MS-PCR:

Methylation-sensitive PCR

MSAP:

Methylation-sensitive amplification polymorphism

CTAB:

Cetyltrimethyl ammonium bromide

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Acknowledgments

This work was financed by the National Natural Science Foundation of China (Grant No. 30971794).

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Authors and Affiliations

  1. Key Laboratory of Maize Biology and Genetic Breeding on Southwest of China, Ministry of Agriculture, Maize Research Institute of Sichuan Agricultural University, Chengdu, Sichuan, 611130, China

    Bo Chen, Yanli Lu, Jing Wang, Suzhi Zhang, Hai Lan, Tingzhao Rong & Moju Cao

  2. Agronomy Department of Xichang College, Xichang, Sichuan, 615013, China

    Bo Chen & Yan Zhang

Authors
  1. Bo Chen
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  2. Yan Zhang
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  3. Yanli Lu
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  4. Jing Wang
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  5. Suzhi Zhang
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  6. Hai Lan
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  7. Tingzhao Rong
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  8. Moju Cao
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Correspondence to Tingzhao Rong or Moju Cao.

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Chen, B., Zhang, Y., Lu, Y. et al. DNA methylation analysis of sterile and fertile CMS-C hybrids and their parents in maize. J. Plant Biochem. Biotechnol. 25, 3–11 (2016). https://doi.org/10.1007/s13562-015-0298-6

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  • DOI: https://doi.org/10.1007/s13562-015-0298-6

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