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Development of a new system for detecting mutation in higher plants and specificity of mutational spectra

高等植物突变检测新系统及突变谱特异性的开发

基本信息

批准号：
16510036
负责人：
TAKIMOTO Koichi
金额：
$ 2.37万
依托单位：
Yamaguchi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16510036/
关键词：
mutation base change plant eukaryote DNA sequence base chang higher plan

项目摘要

Reactive oxygen species, ionizing radiation, UV and various chemicals damage DNA and induce retardation of gene expression, replication and mutation. In plants continuously exposed to UV contained in solar radiation, various types of damages should be produced in genetic substances. These damages has been known to induce mutation. A new system was developed for detection and analysis of mutations occurring on chromosomal DNA in Arabidopsis. The plasmid pML4 carrying the Escherichia coli rpsL gene, a target gene for mutagenesis, was inserted into a shuttle vector to construct a plasmid which could be used for the transformation of plants. pML4 sequences were introduced into Arabidopsis thaliana mediated by Agrobacterium. The pML4 DNA was rescued from transgenic Arabidopsis plant DNA exposed to mutagens, and the plasmids were introduced into Escherichia coli DH10B to detect mutant clones. In this system, any form of inactivation mutation in the rpsL gene can be positively selected since it makes the E.coli cells resistant to streptomycin. We successfully established the assay system of mutation occurring in higher plants. We investigated the mutational specificity in Arabidopsis treated with ethtylmethanesulfonate (EMS).[Results]Mutant frequency of EMS treated plants was 20-fold higher than background mutation. Frameshifts, deletions and sequence substitutions were observed in background mutation. Sequence substitutions have not detected in mouse and fish. GC to AT transitions dominated in EMS-induced mutation. Almost all 3'-purine flanking to target G was A, which was different from animals.

活性氧、电离辐射、紫外线和各种化学物质会损伤 DNA，并导致基因表达、复制和突变迟缓。植物持续暴露于太阳辐射中的紫外线下，其遗传物质会产生各种类型的损害。已知这些损伤会诱发突变。开发了一种新系统，用于检测和分析拟南芥染色体 DNA 上发生的突变。将携带诱变靶基因大肠杆菌rpsL基因的质粒pML4插入穿梭载体中，构建可用于植物转化的质粒。 pML4 序列通过农杆菌介导引入拟南芥。从暴露于诱变剂的转基因拟南芥植物DNA中拯救出pML4 DNA，并将该质粒导入大肠杆菌DH10B中以检测突变克隆。在该系统中，rpsL 基因中任何形式的失活突变都可以被积极选择，因为它使大肠杆菌细胞对链霉素产生抗性。我们成功建立了高等植物突变检测系统。我们研究了乙基甲磺酸(EMS)处理的拟南芥的突变特异性。[结果]EMS处理的植物的突变频率比背景突变高20倍。在背景突变中观察到移码、缺失和序列替换。尚未在小鼠和鱼中检测到序列替换。 GC 到 AT 的转变在 EMS 诱导的突变中占主导地位。目标 G 侧翼的几乎所有 3'-嘌呤都是 A，这与动物不同。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Protection against photooxidative injury of tobacco leaves by 2-alkenal reductase.: Detoxication of lipid peroxide-derived reactive carbonyls

DOI：
10.1104/pp.105.070391
发表时间：
2005-12-01
期刊：
PLANT PHYSIOLOGY
影响因子：
7.4
作者：
Mano, J;Belles-Boix, E;Kushnir, S
通讯作者：
Kushnir, S

Seed deterioration due to high humidity and high temperature is suppressed by extremely low frequency magnetic fields

极低频磁场可抑制因高湿度和高温导致的种子劣化

DOI：
发表时间：
2006
期刊：
Seed Science ＆ Technology 34・1
影响因子：
0
作者：
Jun'ichi Mano;Takeshi Nakahara;Yoshimitsu Torii;Hideki Hirose;Junji Miyakoshi;Koichi Takimoto;Jun'ichi Mano
通讯作者：
Jun'ichi Mano

Actual study for the analysis of the spectrum of mutation occurring in higher plants

高等植物突变谱分析的实际研究

DOI：
发表时间：
2006
期刊：
Radiation Biology Research Communications 41(1)
影响因子：
0
作者：
Yamaguchi Y;Yamada K;Suzuki T;Wu YP;Kita K;Takahashi S;Ichinose M;Suzuki N;自然環境研究センター;Ryouhei Yoshihara
通讯作者：
Ryouhei Yoshihara

Protection against photooxidative injury of tobacco leaves by 2-alkenal reductase. Detoxication of lipidS peroxide-derived reactive carbonyls

2-烯醛还原酶对烟叶光氧化损伤的保护作用。