| Metabolite Profiling of Mature Arabidopsis thaliana Seeds Using Gas Chromatography-Mass Spectrometry (GC-MS)
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Author:
Date:
2016-11-05
[Abstract] Metabolite profiling using gas chromatography-mass spectrometry (GC-MS) permits the annotation and quantification of a relatively wide variety of metabolites, covering a wide range of biochemical groups of metabolites. Lisec et al. (2006) established a method for GC-MS profiling in plants. Based on this protocol, we provide here a detailed GC-MS-based metabolite profiling protocol to identify compounds belonging to several biochemical groups in the primary metabolism of mature Arabidopsis thaliana seeds (Cohen et al., 2014). The protocol uses methoxyamine hydrochloride and N-methyl-N-trimethylsilyltriflouroacetamide (MSTFA) as derivatization reagents, as previous studies indicated these are the most appropriate compounds for profiling of plant ...
[摘要] 使用气相色谱 - 质谱（GC-MS）的代谢物分析允许注释和量化相对多种代谢物，涵盖广泛的代谢物的生物化学组。 Lisec等人。 （2006）建立了一种在植物中进行GC-MS分析的方法。基于该方案，我们在这里提供详细的基于GC-MS的代谢物分析方案以鉴定属于成熟拟南芥种子的主要代谢中的几个生物化学基团的化合物（Cohen等人。，2014）。该方案使用甲氧基胺盐酸盐和N，N-甲基-N，N-三甲基甲硅烷基三氟乙酰胺（MSTFA）作为衍生化试剂，如先前的研究所表明的，这些是用于分析植物代谢物的最合适的化合物。该方案相对快速，提供可重复的结果，并且可以用于简化许多其它类型的植物组织的代谢物，只有很少的修改。在这种情况下，开发种子可以作为研究代谢调节的优秀系统，因为在它们的开发期间，储备化合物的大规模合成在严格的转录调节下发生控制并且与时间上不同的代谢开关相关。
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| Seed Storage Reserve Analysis
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Author:
Date:
2014-10-20
[Abstract] One of the major goals of plant research is to improve crop yield, by for instance increasing seed oil or protein content. Besides this, extensive research is done to change seed fatty acid (FA) composition in order to make vegetable oils more suitable for specific purposes. To determine the effect of genetic changes on seed FA composition, oil, protein and sugar content it’s important to use standardised protocols to compare results between different research groups. Here we describe standardised methods for the analysis of seed FA composition, oil, protein and sugar content.
[摘要] 植物研究的主要目标之一是通过例如增加种子油或蛋白质含量来改善作物产量。 除此之外，还进行了广泛的研究以改变种子脂肪酸（FA）组成，以使植物油更适合于特定目的。 为了确定遗传变化对种子FA组成，油，蛋白质和糖含量的影响，使用标准化方案来比较不同研究组之间的结果是重要的。 在这里我们描述标准化的方法，用于分析种子FA组成，油，蛋白质和糖含量。
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| Metabolite and Fatty Acid Analysis of Yeast Cells and Culture Supernatants
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Author:
Date:
2014-09-05
[Abstract] Metabolite and fatty acid analysis play important roles in evaluating the metabolic state of microorganisms. To examine the growth state and metabolism response of cells to environmental stress or genetic modification, intracellular and extracellular metabolites including fatty acids are usually analyzed to help understand the cellular biochemical changes in microorganisms. In this protocol, gas chromatography-mass spectrometry based analysis was employed to investigate the fatty acids and other metabolites in yeast cells.
[摘要] 代谢物和脂肪酸分析在评估微生物的代谢状态中起重要作用。 为了检查细胞对环境胁迫或遗传修饰的生长状态和代谢反应，通常分析细胞内和细胞外代谢物（包括脂肪酸）以帮助理解微生物中的细胞生物化学变化。 在本协议中，采用基于气相色谱 - 质谱法的分析来研究酵母细胞中的脂肪酸和其他代谢物。
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