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林业科学 ›› 2014, Vol. 50 ›› Issue (3): 99-108.doi: 10.11707/j.1001-7488.20140314

• 论文与研究报告 • 上一篇    下一篇

甲基磺酸甲酯对毛泡桐丛枝病苗DNA甲基化的影响

曹喜兵, 赵振利, 范国强, 邓敏捷, 董焱鹏   

  1. 河南农业大学泡桐研究所 郑州 450002
  • 收稿日期:2013-08-14 修回日期:2013-11-29 出版日期:2014-03-25 发布日期:2014-04-16
  • 基金资助:

    国家自然科学基金项目(30271082;30571496);高等学校博士学科点基金项目(20050466003);中国博士后基金项目(20070410874)。

Effect of Methyl Methanesulphonate on DNA Methylation of Witches’Broom Seedlings of Paulownia tomentosa

Cao Xibing, Zhao Zhenli, Fan Guoqiang, Deng Minjie, Dong Yanpeng   

  1. Institute of Paulownia, Henan Agricultural University Zhengzhou 450002
  • Received:2013-08-14 Revised:2013-11-29 Online:2014-03-25 Published:2014-04-16
  • Contact: 范国强

摘要:

分别采用扩增片段长度多态性(AFLP)和甲基化敏感扩增多态性(MSAP)技术,研究不同浓度甲基磺酸甲酯(MMS)处理毛泡桐丛枝病幼苗后DNA碱基序列及其甲基化的变化,并对发病相关特异DNA片段对应蛋白的功能进行预测。结果表明:健康和丛枝病幼苗及MMS处理后丛枝病幼苗的DNA碱基序列在AFLP水平上相同。随着MMS浓度的升高,处理后丛枝病幼苗的总DNA甲基化水平逐渐升高,但最大浓度(100 mg·L-1)MMS处理后的DNA甲基化水平仍低于健康苗。与毛泡桐丛枝病幼苗相比,MMS处理后丛枝病幼苗的DNA去甲基化多态性皆低于DNA甲基化多态性。序列同源性功能分析结果表明特异DNA甲基化片段对应蛋白功能可能涉及物质和能量代谢、转录和转运、信号转导及致病相关。毛泡桐丛枝病发生可能与其DNA甲基化水平降低和DNA甲基化模式变化密切相关。

关键词: 毛泡桐, 丛枝病, 甲基磺酸甲酯, DNA甲基化, AFLP, MSAP

Abstract:

Changes of the genomic DNA sequences and DNA methylation of P. tomentosa witches' broom seedlings treated with MMS and the specific DNA methylation fragments were analyzed using amplified fragment length polymorphism (AFLP) and methylation sensitive amplification polymorphism (MSAP), respectively. The results showed that there was no difference in DNA base sequences between the healthy seedlings and MMS-treated seedlings at AFLP level. The DNA methylation level of the witches' broom seedlings gradually increased with the increase of MMS concentration, but DNA methylation level of the seedlings treated with the highest concentration (100 mg·L-1)of MMS was still lower than that of the healthy one. Compared with the witches' broom seedlings, DNA demethylation polymorphisms of the MMS-treated seedlings were lower than those of DNA methylation polymorphisms. The specific fragments caused by DNA methylation would be involved in substance and energy metabolism,transcription and transportation, signal transduction and pathogenic factor. Thus the witches' broom was resulted from the decreases of DNA methylation levels and DNA methylation pattern changes of P. tomentosa.

Key words: Paulownia tomentosa, witches' broom, methyl methanesulphonate(MMS), DNA methylation, AFLP, MSAP

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