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首页> 外文期刊>Chinese science bulletin >Patterns of DNA cytosine methylation between haploids and corresponding diploids in rice
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Patterns of DNA cytosine methylation between haploids and corresponding diploids in rice

机译：水稻单倍体与相应二倍体之间DNA胞嘧啶甲基化的模式

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Eighteen pairs of diploid-haploid twin-seedlings were identified and screened out from special rice SARII-628 population. Five pairs of them were selected and randomly designated as A, B, C, D and E. Simple sequence repeats (SSR) analysis showed that there was no difference among 310 sites, which indicated that there was no base mutation on DNA primary structure. DNA methylation plays an important role in gene expression regulation during growth and development stages in eukaryot.es. A modified AFLP technique (methylation-sensitive AFLP, MSAP) was employed to detect the DNA methylation patterns in the 5'-CCGG sites of the five pairs of twin-seedlings. Although no methylation mutation was detected among the five diploids, forty-three methylation mutation sites were found from the corresponding haploids. The MSAP ratios, which were the ratios of MSAP sites to the total amplified sites, in five haploids were 18.79 percent, 19.35 percent, 18.49 percent, 18.45 percent and 18.75 percent, respectively. And corresponding full methylation levels (5'-C~mCGG in double strands) of those haploids were 10.58 percent, 11.3 percent, 10.11 percent, 10.09 percent and 10.34 percent, respectively. Both MSAP and full methylation levels in the five haploids were higher than that of their corresponding diploids, which suggested that hypermethylation occurred in some 5'-CCGG sites. Five types of MASP patterns among the five pairs of twin-seedlings were detected as follows: (1) no changes, methylation levels were the same in both haploids and diploids; (2) demethylation, diploid was methylated but no methylation in the same site in haploid; (3) hypermethylation, the methylation level in haploid was higher than those in diploid; (4) hypomethylation, methylation in haploid was lower than those in diploid; (5) undecided pattern, change trend of methylation levels in haploids was not decided. The bands of 18 sites were reclaimed, then sequenced and searched on website to determine the sites of those sequences on rice chromosomes. The result showed that the methylation mutation involved the whole rice genome and 12 pairs of chromosomes. The mutation was site-related and there were different mutation sites for different haploids. Compared to diploids, the higher methylation level in haploids might be a readjusting reaction to the decrease in ploidy for the sake of survival.
机译：从特殊水稻SARII-628群体中鉴定并筛选出18对二倍体-单倍体双苗。选择了五对，并随机分配为A，B，C，D和E。简单序列重复（SSR）分析显示310个位点之间没有差异，这表明DNA一级结构上没有碱基突变。在真核生物的生长和发育阶段，DNA甲基化在基因表达调控中起着重要作用。采用改良的AFLP技术（对甲基化敏感的AFLP，MSAP）来检测五对双苗的5'-CCGG位点的DNA甲基化模式。尽管在五个二倍体中未检测到甲基化突变，但从相应的单倍体中发现了四个甲基化突变位点。在五个单倍体中，MSAP位点（即MSAP位点与总扩增位点的比率）分别为18.79％，19.35％，18.49％，18.45％和18.75％。这些单倍体的相应全甲基化水平（双链中的5'-C〜mCGG）分别为10.58％，11.3％，10.11％，10.09％和10.34％。五个单倍体的MSAP和全甲基化水平均高于其相应的二倍体，这表明在5'-CCGG位点发生了高甲基化。在五对双子苗中检测出五种类型的MASP模式：（1）单倍体和二倍体均无变化，甲基化水平相同; （2）脱甲基，二倍体被甲基化，但在单倍体的同一位点没有甲基化; （3）高甲基化，单倍体的甲基化水平高于二倍体; （4）低甲基化，单倍体甲基化程度低于二倍体; （5）未确定的模式，未确定单倍体中甲基化水平的变化趋势。回收18个位点的条带，然后在网站上进行测序和搜索，以确定这些序列在水稻染色体上的位点。结果表明，甲基化突变涉及整个水稻基因组和12对染色体。该突变与位点相关，并且对于不同的单倍体存在不同的突变位点。与二倍体相比，单倍体中更高的甲基化水平可能是为了生存而对减少倍数的一种重新调节反应。

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