Clarification of pathway-specific inhibition by Fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies.
We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.
Oikawa
Akira
A
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai 599-8531, Japan.
Nakamura
Yukiko
Y
Ogura
Tomonori
T
Kimura
Atsuko
A
Suzuki
Hideyuki
H
Sakurai
Nozomu
N
Shinbo
Yoko
Y
Shibata
Daisuke
D
Kanaya
Shigehiko
S
Ohta
Daisaku
D
D017360:Arabidopsis / Q000378:metabolism*; D017357:Cyclotrons; D005583:Fourier Analysis; D018525:Germination; D006540:Herbicides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D010641:Phenotype; D036226:Seedlings / Q000378:metabolism
D006540:Herbicides