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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

为了研究硫缺乏对拟南芥mRNA积累谱的影响，利用DNA宏阵列分析了拟南芥约8000个基因的ESTS序列。将生长在对照培养基上的3周大的拟南芥植株转移到无硫酸盐培养基上，培养48小时，分析硫相关代谢物和全球基因表达谱。测定硫酸盐、o -乙酰- l-丝氨酸（OAS）、半胱氨酸和谷胱甘肽（GSH）的浓度，OAS是硫缺乏反应基因的正调节因子。Macroarray分析发现了APR1和sultr1等多个基因；2，缺硫使其mRNA积累量增加。在硫酸盐充足的条件下，用OAS处理的植株也进行了剖面分析。散点图分析结果显示，硫缺乏与OAS处理后的基因表达量变化呈正相关，表明在硫缺乏条件下，多个基因的mRNA积累主要受组织中OAS浓度的控制。叶片和根系缺硫调控的基因组也存在较大差异。植物将氮和硫(S)储存为种子储存蛋白（ssp），以备下一代萌发。ssp的组成根据土壤中硫的有效性而变化，以维持种子中的氮含量。通过蛋白质组学分析，研究了s缺乏和对照条件下拟南芥籽粒蛋白质积累模式的差异。缺s条件下，几种蛋白增加。它们被鉴定为12S ssp、CRA1和推测的十字花素的形式。MALDI-TOFMS分析显示，缺s条件下c端加工未发生。其他在S缺乏下增加的蛋白质被鉴定为CRA1的前体和CRA1的α亚基，以及推定的等电点移位的十字花素。这些结果表明，在s缺乏条件下，ssp经历了一些不同的翻译后裂解和修饰。少