Application of GC-MS for the detection of lipophilic compounds in diverse plant tissues
1 Max-Planck Institute of Molecular Plant Physiology, Am Muehlenberg 1, 14476 Potsdam-Golm, Germany
2 CRA Cereal Research Center, S.S. 16, km 675, 71100 Foggia, Italy
3 Department of Plant Biology, Cornell University, 331 Emerson Hall, Ithaca, New York 14853, USA
4 Institute of Biology/Applied Genetics, Free University of Berlin, Albrecht-Thaer-Weg 6, 14195 Berlin, Germany
5 School of Biological Sciences, Washington State University, PO Box 644236 Pullman, WA 99164-4236, USA
6 Present address: De Ruiter Seeds, Leeuwenhoekweg 52, 2661CZ Bergschenhoek, the Netherlands
Plant Methods 2009, 5:4 doi:10.1186/1746-4811-5-4Published: 24 April 2009
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.