中国农业科学 ›› 2005, Vol. 38 ›› Issue (11): 2301-2307 .

• 园艺 • 上一篇    下一篇

应用MSAP技术对脐橙品种进行DNA甲基化分析

洪 柳,邓秀新   

  • 收稿日期:2005-06-27 修回日期:2005-08-17 出版日期:2005-11-10 发布日期:2005-11-10

Analysis of DNA Methylation in Navel Oranges Based on MSAP Marker

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  • Received:2005-06-27 Revised:2005-08-17 Online:2005-11-10 Published:2005-11-10

摘要: 应用甲基化敏感扩增多态性(MSAP)技术对24个脐橙品种胞嘧啶甲基化模式和程度进行评估。结果表明,脐橙基因组的CCGG序列中检测到4.7%~15.0%的DNA发生甲基化;18对扩增引物有10对显示有多态性,总共得到639条带,其中43条带为甲基化多态性带,平均甲基化多态性(P)比率达6.7%。结果显示DNA甲基化在脐橙中发生频繁,且品种之间的甲基化模式存在较大差异,脐橙CCGG序列中胞嘧啶甲基化外部(15.0%)多于内部(4.7%)。本文首次运用MSAP技术对脐橙品种进行胞嘧啶甲基化分析表明,使用MSAP检测脐橙品种多态性非常有效。

关键词: 脐橙, DNA甲基化, MSAP

Abstract: The extent and pattern of cytosine methylation in 24 navel orange cultivars,genomes were assessed by using the technique of methylation-sensitive amplified polymorphism (MSAP). 4.7%-15.0% DNA methylation events were detected in the CCGG sequence of navel orange genomes. In all, 639 fragments were amplified using 18 pairs of selective primers, and 10 of the 18 primers showed 43 DNA methylation polymorphisms. The average proportion of polymorphism(P)was 6.7%. The results demonstrate DNA methylation events occur frequently in navel oranges;methylation patterns vary among the cultivars, and the external cytosine methylation of the CCGG sequence(15.0%) is more than internal one (4.7%). This paper is the first time to analyse cytosine methylation in navel oranges using MSAP, indicating MSAP technique is efficient for detection of cytosine methylation in navel orange genomes.

Key words: Navel oranges [Citrus sinensis(L.)Osbeck], DNA methylation, MSAP