二斑叶螨不同发育时期基因组DNA甲基化的F-MSAP分析
Using F\|MSAP to investigate genomic DNA methylation in the different developmental stages of Tetranychus urticae
徐敏，杨思霞，赵婧妤，孙荆涛，洪晓月
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作者单位：南京农业大学昆虫学系南京210095
中文关键词：DNA甲基化， 二斑叶螨， F-MSAP
英文关键词：DNA methylation, Tetranychus urticae, FMSAP
中文摘要：DNA甲基化是表观遗传调控的重要机制，在真核生物基因表达调控中发挥重要作用。本研究通过荧光标记的甲基化敏感扩增多态性技术（FMSAP, fluorescencelabeled methylationsensitive amplified polymorphism）对二斑叶螨Tetranychus urticae Koch 4个龄期（卵、幼螨、若螨、成螨）基因组DNA中CCGG位点的胞嘧啶甲基化水平和模式进行分析。研究结果显示3种甲基化模式：无甲基化（Type Ⅰ），半甲基化（Type Ⅱ），全甲基化（Type Ⅲ）在4个龄期均有出现，扩增的总甲基化位点共有641个，其中半甲基化率（Type Ⅱ）均高于全甲基化率（Type Ⅲ），各个龄期的平均总甲基化率（Type Ⅱ + Type Ⅲ）为1601%，平均半甲基化率为1024%，平均全甲基化率为577%。FMSAP分析结果表明不同发育时期的二斑叶螨基因组DNA的甲基化水平和模式存在差异。
英文摘要：
DNA methylation is a key mechanism underlying epigenetic regulation and plays a very important role in regulating gene expression. The level and pattern of cytosine methylation in genomic DNA CCGG sites of the four stages (egg, larva, nymph, adult) of Tetranychus urticae Koch were assessed using the FluorescenceLabeled methylationsensitive amplified polymorphism (FMSAP) technique. The results indicate that all three types of methylation patterns (Type Ⅰ: unmethylated, Type Ⅱ: hemimethylated, Type Ⅲ: full methylated) were observed in all four stages. The total number of amplification methylation sites was 641, in which the hemimethylated ratio was higher than the full methylation ratio across all four stages. The average total methylated ratio (Type Ⅱ + Type Ⅲ), hemimethylated methylated ratio (Type Ⅱ), and full methylation ratio (Type Ⅲ) were 16.01%, 10.24% and 5.77%, respectively. The results of FMSAP analysis demonstrate that the genomic DNA methylation level and pattern were different across the four different developmental stages of T. urticae.