Isolation and analysis of mutants with altered ABA recognition specificity using an ABA antagonist.

使用 ABA 拮抗剂分离和分析 ABA 识别特异性发生改变的突变体。

• 批准号: 12640642
• 负责人: HIRAYAMA Takashi
• 金额: $ 2.62万
• 依托单位: RIKEN (The Institute of Physical and Chemical Reseach)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12640642/
• 关键词: Abscisic acid; mutant; ABA analogues; ABA hypersensitive mutants; HSP90; ethanol hypersensitive mutants; archaea; シロイヌナズナ

In order to identify the ABA receptors, we employed a novel screening using an ABA binding inhibitor to isolate mutants that have altered binding specificity of ABA receptors. We used a ABA analogue, PBI-51, as a ABA binding inhibitor since this chemical has been reported to work as ABA binding inhibitor in Brassica napus. We tried to isolate mutants with two type of screening strategies. One is to search for mutants that seem to recognize PB151 as an ABA binding inhibitor. We isolated several candidates from 500,000 M2 seeds from EMS treated plants. However, these candidates turned out to be ABA insensitive mutants. The other strategy was to isolate mutants that seems recognize PB151 as ABA. In this screen, we isolated 8 independent lines from 400,000 M2 seeds from EMS treated plants. Those candidates show ABA-hypersensitivity, although these are not allelic to known ABA-hypersensitive mutants, eral or ein2.To increase the efficiency to isolate ABA receptor mutants, we thought out the strategy to use aba2 mutants, and isolated several dozens of candidates. ldentification of those mutated genes will surely contribute in our understanding on ABA siganling pathway. In the course to isolate ABA related mutants, we isolated a novel ethanol hypersensitiv mutant, gekol, from Arabidopsis. We identified the GEKO I gene by mapping and found it encoded a novel protein that conserved among plants and archea bacteria. GEKO I may play a pivotal role for plant sp ecific functions.

为了鉴定 ABA 受体，我们采用了一种新的筛选方法，使用 ABA 结合抑制剂来分离改变 ABA 受体结合特异性的突变体。我们使用 ABA 类似物 PBI-51 作为 ABA 结合抑制剂，因为据报道这种化学物质在甘蓝型油菜中可作为 ABA 结合抑制剂。我们尝试用两种筛选策略来分离突变体。一是寻找似乎将 PB151 识别为 ABA 结合抑制剂的突变体。我们从来自 EMS 处理植物的 500,000 M2 种子中分离出了几种候选物。然而，这些候选者被证明是 ABA 不敏感突变体。另一种策略是分离似乎将 PB151 识别为 ABA 的突变体。在此筛选中，我们从 EMS 处理植物的 400,000 M2 种子中分离出 8 个独立品系。这些候选者表现出 ABA 超敏性，尽管它们与已知的 ABA 超敏突变体 eral 或 ein2 并不等位。为了提高分离 ABA 受体突变体的效率，我们想出了使用 aba2 突变体的策略，并分离了几十个候选者。这些突变基因的鉴定必将有助于我们对 ABA 信号通路的理解。在分离ABA相关突变体的过程中，我们从拟南芥中分离出了一种新型的乙醇过敏突变体gekol。我们通过作图鉴定了 GEKO I 基因，并发现它编码了一种在植物和古细菌中保守的新型蛋白质。 GEKO I 可能在植物特定功能中发挥关键作用。

项目成果

Hirayama, T., Alonso, J.M.: "Ethylene Captures a Metal! Metal ions are involved in ethylene perception and signal transduction"Plant & Cell Physiology. 41. 548-555 (2000)

Hirayyama, T., Alonso, J.M.：“乙烯捕获金属！金属离子参与乙烯感知和信号转导”植物

Takahashi, S., Katagiri, T., Hirayama, T., Yamagushi-Shinozaki, K., Shinozaki, K.: "Hyperosmotic stress induces a rapid and transient increase in inositol 1,4,5-trisphosphate independent of abscisic acid in Arabidopsis cell culture"Plant & Cell Physiology

Takahashi, S.、Katagiri, T.、Hirayama, T.、Yamagushi-Shinozaki, K.、Shinozaki, K.：“高渗应激会导致肌醇 1,4,5-三磷酸快速而短暂地增加，与脱落酸无关。

Hirayama, T., Alonso, J.M.: "Ethylene Captures a Metal! Metal ions are involved in ethylene perception and signal transduction"Plant Cell and Physiology. 41. 548-555 (2000)

Hirayama, T.，Alonso, J.M.：“乙烯捕获金属！金属离子参与乙烯感知和信号转导”植物细胞和生理学。

Hirayama, T., & Alonso, J. M: "Ethylene Captures a Metal!Metal ions are involved in ethylene perception and signal transduction"Plant Cell Physiol.,. Vol.41. 548-555 (2000)

平山，T.，

Takahashi, S., Katagiri, K., Hirayama, T., YamaguchiShinozaki, K., Shinozaki, K: "Hyperosmotic stress induces a rapid and transient increase in inositol 1, 4, 5-trisphospate independent of abscisic acid in Arabidopsis cell culture."Plant Cell Physiol.,. V

Takahashi, S.、Katagiri, K.、Hirayama, T.、YamaguchiShinozaki, K.、Shinozaki, K：“在拟南芥细胞培养物中，高渗应激会导致肌醇 1,4,5-三磷酸快速、短暂地增加，与脱落酸无关。