Biosynthesis and distribution of ABA in relation to cellularization, differentiation and storage product accumulation in the barley endosperm

ABA 的生物合成和分布与大麦胚乳细胞化、分化和储存产物积累的关系

• 批准号: 148133760
• 负责人: Professorin Dr. Bettina Hause
• 金额: --
• 依托单位: Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung (IPK)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2009
• 资助国家: 德国
• 起止时间: 2008-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/148133760?language=en
• 关键词: Biosynthesis; distribution; ABA; relation; cellularization

In the previous project, we showed that ABA influences both, cellularization and storage product accumulation of the developing barley endosperm. In this application, we plan to identify (1) tissues producing ABA to regulate seed development, (2) gradients of ABA distribution in developing seeds and (3) regulators of seed development as related to ABA, based on visualization of regions that differ in ABA distribution between wild type and mutant/transgenic seeds, and on subsequent analysis of the respective tissues by micro-methods. To identify ABA-producing tissues, transcripts of ABA biosynthesis will be localized. ABA amounts in different seed parts will be quantified by GC analysis of cryo-microdissectates. Immuno-localization and MALDI-imaging mass spectrometry will be used to show spatial ABA distribution. pH gradients in developing grains will be visualized and correlated to ABA distribution. Bioinformatics tools will be further developed and used to visualize differences in ABA distribution and pH gradients between wild type and mutant/transgenic seeds. To identify new sequences related to ABA distribution and endosperm differentiation, 454 sequencing of the transcriptome of the young endosperm epidermis is planned. Using the sequences, a 44K AGILENT chip will be produced. Regions with different ABA distribution between WT and mutant/transgenic seeds will be micro-dissected and analyzed by transcript and metabolite profiling to identify ABA-related regulatory components and processes.

在之前的研究中，我们发现ABA对发育中的大麦胚乳的细胞化和贮藏产物积累都有影响。在本应用中，我们计划通过可视化野生型和突变/转基因种子之间ABA分布的不同区域，并随后通过显微方法对各自组织进行分析，确定(1)产生ABA调节种子发育的组织，(2)ABA在发育种子中的分布梯度，以及(3)与ABA相关的种子发育调控因子。为了鉴定产生ABA的组织，需要定位ABA生物合成的转录本。种子不同部位的ABA含量将通过气相色谱分析进行定量。免疫定位和maldi成像质谱法将用于显示ABA的空间分布。发育中的颗粒的pH梯度将可视化并与ABA分布相关。生物信息学工具将进一步开发，并用于可视化野生型和突变/转基因种子之间ABA分布和pH梯度的差异。为了确定与ABA分布和胚乳分化相关的新序列，我们计划对幼嫩胚乳表皮的转录组进行454测序。利用这些序列，将生产出44K的AGILENT芯片。通过转录物和代谢物谱分析，对WT和突变/转基因种子之间ABA分布不同的区域进行微观解剖和分析，以确定ABA相关的调控成分和过程。