Analysis of DNA methylation variation in wheat genetic background after alien chromatin introduction based on methylation-sensitive amplification polymorphism
Analysis of DNA methylation variation in wheat genetic background after alien chromatin introduction based on methylation-sensitive amplification polymorphism
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基于甲基化敏感扩增多态性分析小麦遗传背景外源染色质引入后DNA甲基化变异
DOI：
10.1007/s11434-008-0049-3
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发表时间：
2008-01-01
影响因子：
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通讯作者：
Ren ZhengLong
中科院分区：
文献类型：
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作者：
Zhang Yong;Liu ZhaoHui;Ren ZhengLong
利用染色体工程技术将外源种质导入小麦基因组的过程中，会引起受体基因组结构和基因表达的广泛遗传变异。本研究通过对小麦-黑麦染色体易位系及其亲本进行GISH（基因组原位杂交）和AFLP（扩增片段长度多态性）检测不同易位系基因组结构的同一性。结果表明，除了相同的1RS染色体易位外，不同易位系的基因组一级结构差异不明显。基因组DNA甲基化敏感扩增多态性（MSAP）分析表明，易位系的全甲基化位点比例显著增加（CN12, 20.15%; CN17, 20.91%; CN18, 22.42%），而半甲基化位点比例显著降低（CN12, 21.41%; CN17, 23.43%; CN18, 22.42%），而小麦亲本的全甲基化位点比例为16.37%，半甲基化位点比例为25.44%。对小麦-黑麦易位系与小麦亲本的胞嘧啶甲基化模式进行比较分析，鉴定出29种甲基化模式，其中高甲基化模式13种（33.74%），去甲基化模式9种（22.76%），不确定模式7种（4.07%）。在进一步的序列分析中，甲基化模式的改变既影响重复DNA序列，如反转录转座子和串联重复序列，也影响低拷贝DNA。
During the process of alien germplasm introduced into wheat genome by chromosome engineering, extensive genetic variations of genome structure and gene expression in recipient could be induced. In this study, we performed GISH (genome in situ hybridization) and AFLP (amplified fragment length polymorphism) on wheat-rye chromosome translocation lines and their parents to detect the identity in genomic structure of different translocation lines. The results showed that the genome primary structure variations were not obviously detected in different translocation lines except the same 1RS chromosome translocation. Methylation sensitive amplification polymorphism (MSAP) analyses on genomic DNA showed that the ratios of fully-methylated sites were significantly increased in translocation lines (CN12, 20.15%; CN17, 20.91%; CN18, 22.42%), but the ratios of hemimethylated sites were significantly lowered (CN12, 21.41%; CN17, 23.43%; CN18, 22.42%), whereas 16.37% were fully-methylated and 25.44% were hemimethylated in case of their wheat parent. Twenty-nine classes of methylation patterns were identified in a comparative assay of cytosine methylation patterns between wheat-rye translocation lines and their wheat parent, including 13 hypermethylation patterns (33.74%), 9 demethylation patterns (22.76%) and 7 uncertain patterns (4.07%). In further sequence analysis, the alterations of methylation pattern affected both repetitive DNA sequences, such as retrotransposons and tandem repetitive sequences, and low-copy DNA.