黑麦基因组DNA甲基化修饰位点的MSAP分析  

Analysis on Genomic DNA Methylation Modification of Rye by Methylation-sensitive Amplification Polymorphism

作　　者：张勇[1,2] 邓科君[1] 张韬[1] 彭金华[1] 周建平[1] 任正隆[1,2] 

机构地区：[1]电子科技大学生命科学与技术学院,四川成都610054 [2]四川农业大学植物遗传育种省级重点实验室,四川雅安625014

出　　处：《麦类作物学报》2009年第4期559-564,共6页Journal of Triticeae Crops

基　　金：国家自然科学基金重点项目(30730065);国家博士后科学基金项目(20070411158);电子科技大学青年基金项目

摘　　要：为了获得黑麦基因组DNA甲基化修饰水平、模式及位点等表观遗传信息,采用EcoRⅠ和HpaⅡ/MspⅠ双酶切建立适合于黑麦基因组的"甲基化敏感扩增多态性"(Methylation sensitive amplification polymor-phism,MSAP)分析体系,在全基因组水平检测黑麦DNA甲基化修饰位点。以12对MSAP引物进行选择性扩增,共检测到甲基化修饰位点226个,"CCGG/GGCC"位点甲基化修饰比例为51.72%。对部分黑麦基因组甲基化修饰位点进行回收,最终分离了22条存在甲基化修饰的基因组DNA序列。BLAST比对分析结果表明,黑麦基因组中包括转座子序列、散在重复序列以及单拷贝蛋白质编码序列在内的多种类型DNA序列中均存在DNA甲基化修饰现象。同时发现,在甲基化检出序列中都存在明显的"CpG"二核苷酸成簇富集现象,这些区域分布与MSAP分析结果相一致。在此基础上,对应用MSAP技术分离黑麦基因组DNA甲基化修饰位点的有效性以及黑麦基因组序列中DNA甲基化修饰潜在位点分布特征和生物意义进行了讨论。In this study, the double digestion of EcoR Ⅰ and Hpa Ⅱ/Msp Ⅰwas used to construct the high-density rye （Secale cereale） genomic methylation-sensitive amplification polymorphism fingerprint and detected the methylated sites at the whole genome level. By using 12 pairs of selective primers, a total of 226 methylated sites was detected, which represent 51. 72% ratio of methylation modification at ‘CCGG/GGCC＇ site in rye genome. Some of methylated sites were extracted and sequenced. Further sequence analysis showed the alterations of methylation pattern affected both repetitive DNA sequences, such as retrotransposons and dispersed repetitive sequences, and low-copy protein coding sequences. The region of CpG rich cluster with typical CpG island sequence features was also detected in the sequence of rye methylated sites. Based on these experimental evidences, the effectiveness to isolate rye methylated site by MSAP and the distribution features of CpG island like sequence in rye genome were discussed.

关 键 词：黑麦 表观遗传 DNA甲基化 MSAP 

分 类 号：S512.5[农业科学—作物学] S330