Analysis of DNA methylation in Arabidopsis thaliana based on methylation-sensitive AFLP markers
Analysis of DNA methylation in Arabidopsis thaliana based on methylation-sensitive AFLP markers
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DOI：
10.1007/s00438-002-0772-4
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发表时间：
2002-12-01
影响因子：
3.1
通讯作者：
Martínez-Zapater, JM
中科院分区：
文献类型：
--
作者：
Cervera, MT;Ruiz-García, L;Martínez-Zapater, JM
使用对其识别位点甲基化敏感性不同的限制酶同裂酶进行 AFLP 分析，已用于分析拟南芥中匿名 CCGG 序列的甲基化状态。该技术经过修改以提高指纹的质量并可视化大量可评分的片段。扩增片段的测序表明检测通常与同裂酶敏感的胞嘧啶的非甲基化有关。不同生态型中 EcoRI/HpaII 和 EcoRI/MspI 模式的比较表明，35-43% 的 CCGG 位点被同裂酶差异消化。有趣的是，同一生态型不同植物之间的消化模式高度保守，生态型内甲基化敏感多态性比例小于1%。然而，对属于不同生态型的样品之间的甲基化模式进行两两比较，发现高达 34% 的甲基化敏感多态性存在差异。基于甲基化不敏感或甲基化敏感多态性的生态型间相似性矩阵之间缺乏相关性表明，无论调节甲基化的机制是什么，它们与核苷酸序列变异无关。
AFLP analysis using restriction enzyme isoschizomers that differ in their sensitivity to methylation of their recognition sites has been used to analyse the methylation state of anonymous CCGG sequences in Arabidopsis thaliana. The technique was modified to improve the quality of fingerprints and to visualise larger numbers of scorable fragments. Sequencing of amplified fragments indicated that detection was generally associated with non-methylation of the cytosine to which the isoschizomer is sensitive. Comparison of EcoRI/HpaII and EcoRI/MspI patterns in different ecotypes revealed that 35-43% of CCGG sites were differentially digested by the isoschizomers. Interestingly, the pattern of digestion among different plants belonging to the same ecotype is highly conserved, with the rate of intra-ecotype methylation-sensitive polymorphisms being less than 1%. However, pairwise comparisons of methylation patterns between samples belonging to different ecotypes revealed differences in up to 34% of the methylation-sensitive polymorphisms. The lack of correlation between interecotype similarity matrices based on methylation-insensitive or methylation-sensitive polymorphisms suggests that whatever the mechanisms regulating methylation may be, they are not related to nucleotide sequence variation.