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，我们使用ABA结合抑制剂进行了新的筛选，以分离已改变ABA受体特异性的突变体。我们使用ABA类似物PBI-51作为ABA结合抑制剂，因为据报道该化学物质在甘蓝乳剂中起作用为ABA结合抑制剂。我们试图通过两种类型的筛选策略隔离突变体。一种是寻找似乎识别PB151是ABA结合抑制剂的突变体。我们从EMS处理的植物中分离了500,000平方米种子的几个候选物。但是，这些候选人原来是ABA不敏感的突变体。另一种策略是隔离似乎将PB151识别为ABA的突变体。在此屏幕中，我们从EMS处理的植物中分离出400,000平方米种子的8条独立线。这些候选者表现出ABA - 高敏感性，尽管这些候选者并非与已知的ABA - 甲状腺敏感突变体或EIN。这些突变基因的l固定肯定会为我们对ABA Siganling途径的理解做出贡献。在隔离ABA相关的突变体的过程中，我们将一种新型的乙醇超敏突变体Gekol与拟南芥分离出来。我们通过映射鉴定了Geko I基因，并发现它编码了一种新型蛋白质，该蛋白在植物和古细菌中保守。 Geko I可能会在植物SP ECIFIC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。