Max-Planck Institute of Molecular Plant Physiology, Am Muehlenberg 1, 14476 Potsdam-Golm, Germany.
Plant Methods. 2009 Apr 24;5:4. doi: 10.1186/1746-4811-5-4.
The concept of metabolite profiling has been around for decades and technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. However, studies are generally confined to polar compounds alone. Here we describe a simple method for lipophilic compounds analysis in various plant tissues.
We choose the same preparative and instrumental platform for lipophilic profiling as that we routinely use for polar metabolites measurements. The method was validated in terms of linearity, carryover, reproducibility and recovery rates, as well as using various plant tissues.As a first case study we present metabolic profiling of Arabidopsis root and shoot tissue of wild type (C24) and mutant (rsr4-1) plants deficient on vitamin B6. We found significant alterations in lipid constituent contents, especially in the roots, which were characterised by dramatic increases in several fatty acids, thus providing further hint for the role of pyridoxine in oxidative stress and lipid peroxidation.The second example is the lipophilic profiling of red and green tomato fruit cuticles of wild type (Alisa Craig) and the DFD (delayed fruit deterioration) mutant, which we compared and contrasted with the more focused wax analysis of these plants reported before.
We can rapidly and reliably detect and quantify over 40 lipophilic metabolites including fatty acids, fatty alcohols, alkanes, sterols and tocopherols. The method presented here affords a simple and rapid, yet robust complement to previously validated methods of polar metabolite profiling by gas-chromatography mass-spectrometry.
代谢物分析的概念已经存在了几十年,现在的技术创新使其能够在数量上对大量代谢物进行大规模分析,无论是测量的代谢物数量还是进行的实验数量。然而,这些研究通常仅限于极性化合物。在这里,我们描述了一种简单的方法,用于分析各种植物组织中的亲脂性化合物。
我们选择了与我们常规用于测量极性代谢物的相同的制备和仪器平台来进行亲脂性分析。该方法在线性、携带、重现性和回收率方面进行了验证,同时还使用了各种植物组织。作为第一个案例研究,我们展示了野生型(C24)和突变型(rsr4-1)植物的拟南芥根和茎组织的代谢物分析,这些植物缺乏维生素 B6。我们发现脂质成分含量有显著变化,特别是在根部,几种脂肪酸的含量显著增加,这进一步提示吡哆醇在氧化应激和脂质过氧化中的作用。第二个例子是野生型(Alisa Craig)和 DFD(延迟果实腐烂)突变体的红番茄和绿番茄果皮的亲脂性分析,我们对这些植物进行了比较,并与之前报道的这些植物的更集中的蜡分析进行了对比。
我们可以快速、可靠地检测和定量超过 40 种亲脂性代谢物,包括脂肪酸、脂肪醇、烷烃、甾醇和生育酚。这里介绍的方法为气相色谱-质谱法验证的极性代谢物分析提供了一种简单、快速但稳健的补充方法。