SE151:/S1

Sample Set Information

ID	TSE1306
Title	Plant lipidomics based on hydrophilic interaction chromatography coupled to ion trap time-of-flight mass spectrometry.
Description	Plants synthesize a wide range of hydrophobic compounds, generally known as lipids. Here, we report an application of liquid chromatography ion trap time-of-flight mass spectrometry (LC-IT-TOF-MS) for plant lipidomics. Using hydrophilic interaction chromatography (HILIC) for class separation, typical membrane lipids including glycerolipids, steryl glucosides and glucosylceramides, and hydrophobic plant secondary metabolites such as saponins were analyzed simultaneously. By this method, we annotated approximately 100 molecules from Arabidopsis thaliana. To demonstrate the application of this method to biological study, we analyzed Arabidopsis mutant trigalactosyldiacylglycerol3 (tgd3), which has a complex metabolic phenotype including the accumulation of unusual forms of galactolipids. Lipid profiling by LC-MS revealed that tgd3 accumulated an unusual form of digalactosyldiacylglycerol, annotated as Gal(β1 → 6)βGalDG. The compositional difference between normal and unusual forms of digalactosyldiacylglycerol was detected by this method. In addition, we analyzed well-known Arabidopsis mutants ats1-1, fad6-1, and fad7-2, which are also disrupted in lipid metabolic genes. Untargeted lipidome analysis coupled with multivariate analysis clearly discriminated the mutants and their distinctive metabolites. These results indicated that HILIC-MS is an efficient method for plant lipidomics.
Authors	Okazaki Y, Kamide Y, Hirai MY, Saito K.
Reference	Metabolomics. 2013 Mar;9(Suppl 1):121-131. Epub 2011 May 31.
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PubMed

Sample Information

ID	S1
Title	Arabidopsis thaliana
Organism - Scientific Name	Arabidopsis thaliana
Organism - ID	NCBI taxonomy:3702
Compound - ID
Compound - Source
Preparation	Seeds of A. thaliana mutants (SALK_040335 for tgd3, CS200 for ats1-1, CS207 for fad6-1, and CS8042 for fad7-2) were obtained from the ABRC (Ohio, USA). The T-DNA insertion site in tgd3 was confirmed by sequencing of PCR fragments at the left border of the T-DNA amplified using LBa1 (5′-TGGTTCACGTAGTGGGCCATCG-3′) and gene-specific primer (5′-TGCTTCAGTGGTATGTCATGTGAAAGTAT-3′).
Sample Preparation Details ID	SS1
Comment

Sample Preparation Details Information

ID	SS1
Title	Growth conditions
Description	Arabidopsis plants were grown on agar-solidified Murashige-Skoog medium containing 1% (w/v) sucrose and MS vitamin at 22°C under a 16-h-light/8-h-dark cycle for 18 days. Plant tissues were harvested 6 h after the onset of the light phase, frozen in liquid nitrogen, and stored at −80°C until lipid extraction.
Comment_of_details