Abstract
BackgroundThe 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.ResultsWe 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.ConclusionWe 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.
Highlights
The concept of metabolite profiling has been around for decades and technical innovations are enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out
We choose the same preparative and instrumental platform for lipophilic profiling as that we routinely use for polar metabolites measurements
As a first case study we present metabolic profiling of Arabidopsis root and shoot tissue of wild type (C24) and mutant plants deficient on vitamin B6
Summary
The concept of metabolite profiling has been around for decades and technical innovations are enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. Until relatively recently only a limited number of plant research laboratories had access to gaschromatography mass-spectrometry instrumentation, such machines are increasingly becoming more commonplace. In the medicinal field the majority of studies have arguably been focussed on development of metabolite profiling as a diagnostic tool. In this field impressive examples have been provided by the discovery of markers for coronary heart disease and atherosclerosis [7,11]. Early plant studies focussed in this area (see for example [12,13]), a great deal of research is currently carried out at a more mechanistic level often encompassing other post-genomic tools. Recent examples of note in this direction are the development of combined transcript-metabolite networks for aiding functional gene annotation [14,15] and studies aimed at uncovering the genetic basis of metabolic regulation [16,17,18,19,20]
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