Molecular regulation of sulfur transport, assimilation and conversion in storage function in plants

植物储存功能中硫运输、同化和转化的分子调控

基本信息

批准号：
12138202
负责人：
SAITO Kazuki
金额：
$ 20.67万
依托单位：
CHIBA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2003
项目状态：
已结题

项目摘要

To investigate the changes in profiles of mRNA accumulation in response to sulfur deficiency, Arabidopsis thaliana ESTS corresponding to approximately 8000 genes were analyzed using DNA macroarray. Three-week-old Arabidopsis plants grown on control medium were transferred to a sulfate-free medium and grown for 48 h for the analyses of sulfur-related metabolites and global gene exression profiles. Concentrations of sulfate, O-acetyl-L-serine (OAS), a positive regulator of sulfur deficiency-responsive genes, cysteine and glutathione (GSH) were determined. Macroarray analysis revealed a number of genes, including APR1 and sultr1 ; 2, whose mRNA accumulation was increased by sulfur deficiency. Profiling was also carried with plants treated with OAS under sulfate-sufficient condition. Scatter plot analysis revealed a positive correlation between the changes of expression levels by sulfur deficiency and by OAS treatment among the clones tested, suggesting that mRNA accumulation of a number o … More f genes under sulfur deficiency is mainly controlled by OAS concentrations in tissues. It was also revealed that the sets of genes regulated under sulfur deficiency in leaves and roots differ considerably. Plants reserve nitrogen and sulfur (S) as seed storage proteins (SSPs) for germination of next generation. Composition of SSPs changes according to sulfur availability in soil to maintain nitrogen contents in seeds. We examined the difference of protein accumulation patterns in Arabidopsis seeds harvested under S-deficient and control condition by proteome analysis. Several proteins increased under S-deficient condition. They were identified forms of the 12S SSPs, CRA1 and putative cruciferin. MALDI-TOFMS analyses showed the C-terminal processing did not occurred under S-deficient condition. Other proteins that increased under S deficiency were identified as a precursor of CRA1 and alpha-subunits of CRA1 and putative cruciferin whose isoelectric points were shifted. These results suggested SSPs undergo some different post-translational cleavages and modifications under S-deficient condition. Less

为了研究响应硫缺乏的 mRNA 积累谱的变化，使用 DNA 宏阵列分析了对应于大约 8000 个基因的拟南芥 ESTS，将在对照培养基上生长的三周龄拟南芥植物转移到无硫酸盐培养基中并生长。 48 小时，用于分析硫相关代谢物和整体基因表达谱。微阵列分析发现，O-乙酰-L-丝氨酸（OAS）是硫缺乏反应基因、半胱氨酸和谷胱甘肽（GSH）的正调节因子，其中包括APR1和sultr1 2，其mRNA积累。还对硫酸盐充足条件下用 OAS 处理的植物进行了分析，结果显示这些变化之间呈正相关。在测试的克隆中，硫缺乏和 OAS 处理对表达水平的影响，表明硫缺乏下许多基因的 mRNA 积累主要受组织中 OAS 浓度的控制。还揭示了在硫缺乏下受调节的基因组。叶子和根部缺硫的情况差异很大。植物将氮和硫（S）保留为种子储存蛋白（SSP），用于下一代的萌发。SSP 的组成根据土壤中的硫可用性而变化。我们通过蛋白质组分析检查了在 S 缺陷和对照条件下收获的拟南芥种子中蛋白质积累模式的差异，它们被鉴定为 12S SSP、CRA1 和假定的十字花科素。 MALDI-TOFMS 分析表明，在 S 缺陷条件下，C 末端加工不会发生，在 S 缺陷条件下增加的其他蛋白质被鉴定为 CRA1 和 α 亚基的前体。 CRA1 和假定的十字花科素的等电点发生了变化，这些结果表明 SSP 在 S 缺陷条件下经历了一些不同的翻译后裂解和修饰。