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Genetic factors responsible for the mobility of mPing in the rice genome

导致水稻基因组中 mPing 迁移的遗传因素

基本信息

批准号：
15380006
负责人：
OKUMOTO Yutaka
金额：
$ 9.09万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380006/
关键词：
Rice Transposon MITEs mechanism of transposition Mutation Rurm1 mPing MITE 転移

项目摘要

In a mutant line IM294 (slender glume mutant) which harbors Rurm1 (Rice Ubiquitine Related Modifier-1) allele inactivated by mPing insertion shows high excision frequency (ca.1%) of mPing from mutant Rurm1 allele (Rurm1^m). Expression levels of two ORFs of Ping were also high in IM294 comparing to the original variety Gimbozu. To clarify the relation between the function of Rurm1 and Ping activity under the same genetic background, the mutant allele of Rurm1 was introduced into Gimbozu background through recurrent backcrossing. Also, a pair of non-slender glume line (Rurm1^+/Rrum1^+) and slender glume line (Rurm1^m/Rurm1^m) derived from offspring of the non-slender glume plant (Rurm1^+/Rurm1^m) segregated among IM294 line were used. Comparing the Gimbozu and its isogenic line and sib lines, inactivation of Rurm1 with mPing insertion enhanced the expression levels of two ORFs of Ping. On the other hand, significant change in the expression level of Pong was not observed. These indicate that the inactivation of Rurm1 promotes the mPing excision by enhancing the expression of Ping's ORFs. Though, the inactivation of transposon is generally regulated by methylation, it is quite unlikely that RURM1 protein directly regulate the methylation of DNA. Effect of the inactivation of Rurm1 allele was also analyzed using 22k-micro array (Agilent Co.,). Inactivation of Rurm1 upregulate the expression of protein biosynthesis related genes in ribosome and nucleus. It also downregulated the expression of genes related to metabolism and physiological process. These indicate that the function of Rurm1 is related to transcription of vast number of genes related to the physiological process. Upregulation of protein synthesis related genes indicates the compensation to growth retardation of plants. Upregulation of genes related to DNA-repair process was also observed indicating the frequent DNA double strands break induced by mPing excision in IM294.

在含有Rurm 1（Rice Ubiquitine Related Modifier-1）等位基因的突变系IM 294（细长颖突变体）中，mPing插入失活的突变体Rurm 1等位基因（Rurm1^m）的mPing切除频率高（约1%）。Ping的两个ORF在IM 294中的表达量也高于原品种金宝莲。为了阐明Rurm 1基因的功能与Ping活性在相同遗传背景下的关系，通过轮回回交将Rurm 1基因的突变等位基因导入金保津背景。此外，还使用了一对非细长颖片系（Rurm 1 ^+/Rrum 1 ^+）和细长颖片系（Rurm 1 ^m/Rurm 1 ^m），它们来自在IM 294系中分离的非细长颖片植株（Rurm 1 ^+/Rurm 1 ^m）的后代。比较金保津及其同基因系和同胞系，失活的Rurm 1与mPing插入提高了Ping的两个ORF的表达水平。另一方面，未观察到Pong表达水平的显著变化。这表明Rurm 1的失活通过增强Ping's ORF的表达来促进mPing的切除。虽然转座子的失活通常受甲基化的调控，但RURM 1蛋白不太可能直接调控DNA的甲基化。还使用22 k-微阵列（Agilent Co.，）分析Rurm 1等位基因失活的影响。Rurm 1的失活可上调核糖体和细胞核中蛋白质生物合成相关基因的表达。它还下调与代谢和生理过程相关的基因的表达。这表明Rurm 1的功能与大量与生理过程相关的基因的转录有关。蛋白质合成相关基因的上调表明植物对生长迟缓的补偿作用。还观察到与DNA修复过程相关的基因的上调，表明在IM 294中由mPing切除诱导的频繁的DNA双链断裂。