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Patterns of DNA cytosine methylation between haploids and corresponding diploids in rice

ISSN号：1002-0470
期刊名称：《高技术通讯》
时间：0
分类：N[自然科学总论]
作者机构：[1]Rice Research Institute of Sichuan Agricultural University, Wenjiang 611130, Sichuan, [2]Key Laboratory of Southwest Crop Genetic Resources and Improvement, Ministry of Education, Sichuan Agricultural University, Ya'an625014, China
相关基金：Acknowledgements This work was supported by the Program for Changjiang Scholars and Innovative Research Team in University （Grant No. IRT0453）.

作者： ZHANG Hongyu[1,2] PENG Hai[1,2] LI Yun[1,2] XU Peizhou[1,2] WANG Xudong[1,2] WU Xianjun[1,2]

关键词：米饭, 成双幼苗, 双, HAPLOID, DNA, 甲基化, MSAP, rice, twin-seedling, diploid, haploid, DNA methylation,MSAP.

中文摘要：

十八 diploid-haploid 成双幼苗从特殊米饭 SARII-628 外面被识别并且屏蔽人口。五他们被选择并且随机指定了为 A， B， C， D 和 E。简单顺序重复(SSR ) 分析证明在 310 个地点之中没有差别，它显示 DNA 主要结构上没有基础变化。DNA 甲基化在 eukaryot.es 在生长和开发阶段期间在基因表达式规定起一个重要作用。一种修改 AFLP 技术(methylation 敏感的 AFLP， MSAP ) 被采用在五对 oftwin 幼苗的 5''-CCGG 地点检测 DNA 甲基化模式。尽管没有甲基化变化在五 diploids 之中被检测， forty-threemethylation 变化地点从相应 haploids 被发现。在五 haploids 的 MSAP 比率，是到全部的放大地点的 MSAP 地点的比率，是 18.79% ， 19.35percent， 18.49% ， 18.45% 和 18.75% 分别地。并且那些 haploids 的相应 fullmethylation 层次(在两倍海滨的 5''-C ~ mCGG ) 分别地是 10.58% ， 11.3% ， 10.11% ， 10.09% 和 10.34% 。在五 haploids 的 MSAP 和完整的甲基化层次比他们的相应 diploids 的高，它建议 thathypermethylation 发生在一些 5''-CCGG 地点。在五对 oftwin 幼苗之中的 MASP 模式的五种类型如下被检测：(1 ) 不变化，甲基化层次是在 bothhaploids 和 diploids 的一样；(2 ) 脱甲基作用，双在在 haploid 的一样的地点是 methylated 而是没有甲基化；(3 ) hypermethylation，在 haploid 的甲基化水平比那些在高双；(4 ) hypomethylation，在 haploid 的甲基化是比那些在低的双；(5 ) 未定的模式，在 haploids 的甲基化层次的变化趋势没被决定。18 个地点的乐队在决定米饭染色体上的那些序列的地点的网站上被回收，然后定序并且寻找。结果证明甲基化变化包含了整个米饭染色体和染色体的 12pairs。变化是地点相关的，为不同 haploids 有不同变化地点。比作 diploids，在 haploids 的更高的甲基化水平可能是对在为幸存的缘故的 ploidy 的减少的再整理的反应。

英文摘要：

Eighteen pairs of diploid-haploid twinseedlings 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 eukaryotes. 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%, 19.35%, 18.49%, 18.45% and 18.75%, respectively. And corresponding full methylation levels （5＇-crnCGG in double strands） of those haploids were 10.58%, 11.3%, 10.11%, 10.09% and 10.34%, 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 muta

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