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Capture of transposons utilizing mutable chlorophyll mutant in rice and tomato

利用水稻和番茄中可变叶绿素突变体捕获转座子

基本信息

批准号：
10556002
负责人：
MAEKAWA Masahiko
金额：
$ 7.94万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10556002/
关键词：
mutability variegation chlorophyll mutant transposition transposon rice tomato

项目摘要

For isolating a new transposon in higher plants, the selection of mutants resistant to hydroxyproline (a proline analog) was conducted, because some mutants resistant to an amino acid analog have been controlled by curious genetic behavior. However, through the study no evidence for transposable elements was obtained. (Hasegawa).Unlike most other cut-and-paste type transposons, the transposition frequency of Tam3 in Antirrhinum is tightly controlled by temperature : Tam3 transposes rarely at 25 ℃, but much more frequently at 15 ℃. We found that temperature shift induced a remarkable change of the methylation state of the Tam3 end regions : higher temperature resulted in hyper-methylation, while lower temperature resulted in reduced methylation (kishima).We carried out PCR with primers specific to the TIRs of a 1.3-kb transposon-like element Lyt1 from tomato to search for new members of the Lyt family. Among the amplified products, one fragment had a 2.7-kb insertion in a sequence corre … More sponding to Lyt1 and the putative Lyt element containing this sequence was termed Lyt2. The copy number of Lyt2-related sequences in the tomato genome was estimated to be approximately 20 by Southern blot analysis (Ogura and Masuda).The mutable slender glume gene (slg), a recessive mutant gene induced from rice variety Gimbozu, was assumed to be controlled by a newly found transposon (named Sairyu, meaning slender glume in Japanese) belongs to MITEs family in rice, we suggest that slg is inactivated Rurm1 due to the insertion of Sairyu and that reverse mutation of slg is ascribed to the reactivation of Rurm1 caused by the deletion of Sairyu (Okumoto).An Arabidopsis variegated mutant was isolated by T-DNA tagging. Cloning and molecular characterization of the VAR2 locus revealed that it potentially encodes a chloroplastic homologue of FtsH, an ATP-dependent metalloprotease that belongs to a large protein family involved in various cellular functions (Sakamoto). A mutable chlorophyll mutant was induced from stable chlorophyll mutants by carbon ion beam irradiation. This mutability of the induced mutant was inherited, suggesting that cryptic silent transposon might be reactivated by ion beam irradiation (Maekawa and Tanaka). Less

为了在高等植物中分离新的转座子，对羟脯氨酸（脯氨酸类似物）的抗性突变体进行了选择，因为一些对氨基酸类似物的抗性突变体受到奇怪的遗传行为的控制。然而，通过研究没有获得转座因子存在的证据。(长谷川)。与大多数其他剪切-粘贴型转座子不同，Antirrhinum中Tam3的转座子频率受到温度的严格控制：Tam3在25℃时很少转座子，但在15℃时转座子频率高得多。我们发现温度变化导致Tam3端区的甲基化状态发生显著变化：温度升高导致超甲基化，而温度降低导致甲基化降低（kishima）。我们利用番茄中一个1.3 kb转座子样元件Lyt1的TIRs特异引物进行PCR，寻找Lyt家族的新成员。在扩增产物中，有一个片段在序列中心插入了2.7 kb的片段，更多的是与Lyt1应答，含有该序列的Lyt元件被称为Lyt2。通过Southern blot分析估计番茄基因组中lyt2相关序列的拷贝数约为20个（Ogura和Masuda）。可变细长颖片基因（slg）是水稻品种金波津诱导的一种隐性突变基因，被认为是由一个新发现的水稻螨属转座子（Sairyu，日语中意为细长颖片）控制，我们认为slg是由于Sairyu的插入而使Rurm1失活，而slg的反向突变则是由于Sairyu （Okumoto）的缺失而使Rurm1重新激活。利用T-DNA标记技术分离到一个拟南芥杂交种突变体。VAR2基因座的克隆和分子表征表明，它可能编码FtsH的叶绿体同源物，FtsH是一种atp依赖性金属蛋白酶，属于一个参与多种细胞功能的大蛋白家族（Sakamoto）。以稳定的叶绿素突变体为基础，用碳离子束辐照诱导出一个突变体。诱导突变体的这种易变性是遗传的，这表明隐蔽性沉默转座子可能被离子束照射重新激活（Maekawa和Tanaka）。少