Journal of Integrative Agriculture ›› 2016, Vol. 15 ›› Issue (06): 1228-1238.DOI: 10.1016/S2095-3119(15)61277-1

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Analysis of cytosine methylation in early generations of resynthesized Brassica napus

RAN Li-ping, FANG Ting-ting, RONG Hao, JIANG Jin-jin, FANG Yu-jie, WANG You-ping   

  1. Jiangsu Provincial Key Laboratory of Crop Genetics and Physiology, Yangzhou University, Yangzhou 225009, P.R.China
  • 收稿日期:2015-07-01 出版日期:2016-06-01 发布日期:2016-06-06
  • 通讯作者: WANG You-ping, Tel: +86-514-87997303, Fax: +86-514-87991747, E-mail: wangyp@yzu.edu.cn

Analysis of cytosine methylation in early generations of resynthesized Brassica napus

RAN Li-ping, FANG Ting-ting, RONG Hao, JIANG Jin-jin, FANG Yu-jie, WANG You-ping   

  1. Jiangsu Provincial Key Laboratory of Crop Genetics and Physiology, Yangzhou University, Yangzhou 225009, P.R.China
  • Received:2015-07-01 Online:2016-06-01 Published:2016-06-06
  • Contact: WANG You-ping, Tel: +86-514-87997303, Fax: +86-514-87991747, E-mail: wangyp@yzu.edu.cn
  • Supported by:

    This study was supported by the National Key Basic Research Program of China (2015CB150201), the National Natural Science Foundation of China (NSFC, 31330057, 31401414), the Priority Academic Program Development of Jiangsu Higher Education Institutions of China, the Jiangsu Province Science Foundation, China (BK20140478, 14KJB210008), and the Jiangsu Province Graduate Innovation Fund, China (KYZZ15_0367).

Abstract:     DNA methylation, an important epigenetic modification, serves as a key function in the polyploidization of numerous crops. In this study, early generations of resynthesized Brassica napus (F1, S1–S3), ancestral parents B. rapa and B. oleracea were analyzed to characterize their DNA methylation status during polyploidization, applying DNA methylation-sensitive amplification polymorphism (MSAP) and high-performance liquid chromatography methods. In F1, 53.4% fragments were inherited from both A- and C-genomes. Besides, 5.04 and 8.87% fragments in F1 were inherited from A- and C- genome, respectively. 5.85 and 0.8% fragments were newly appeared and disappeared in resynthesized B. napus, respectively. 13.1% of these gene sites were identified with methylation changes in F1, namely, hypermethylation (7.86%) and hypomethylation (5.24%). The lowest methylation status was detected in F1 (38.7%) compared with in S1–S3. In S3, 40.32% genes were methylated according to MSAP analysis. Sequencing of methylated fragments indicated that genes involved in multiple biological processes were modified, including transcription factors, protein modification, and transporters. Expression ananlysis of DNA methyltransferase 1 and DNA methyltransferase chromomethylase 3 in different materials was consistent to the DNA methylation status. These results can generally facilitate dissection of how DNA methylation contributes to genetic stability and improvement of B. napus during polyploidization.

Key words: resynthesized Brassica napus ,  DNA methylation ,  epigenetics ,  polyploidization