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Sample Set Information

ID TSE1344
Title Rice-Arabidopsis FOX line screening with FT-NIR-based fingerprinting for GC-TOF/MS-based metabolite profiling
Description The full-length cDNA over-expressing (FOX) gene hunting system is useful for genome-wide gain-of-function analysis. The screening of FOX lines requires a high-throughput metabolomic method that can detect a wide range of metabolites. Fourier transform-near-infrared (FT-NIR) spectroscopy in combination with the chemometric approach has been used to analyze metabolite fingerprints. Since FT-NIR spectroscopy can be used to analyze a solid sample without destructive extraction, this technique enables untargeted analysis and high-throughput screening focusing on the alteration of metabolite composition. We performed non-destructive FT-NIR-based fingerprinting to screen seed samples of 3000 rice-Arabidopsis FOX lines; the samples were obtained from transgenic Arabidopsis thaliana lines that overexpressed rice full-length cDNA. Subsequently, the candidate lines exhibiting alteration in their metabolite fingerprints were analyzed by gas chromatography-time-of-flight/mass spectrometry (GC-TOF/MS) in order to assess their metabolite profiles. Finally, multivariate regression using orthogonal projections to latent structures (O2PLS) was used to elucidate the predictive metabolites obtained in FT-NIR analysis by integration of the datasets obtained from FT-NIR and GC-TOF/MS analyses. FT-NIR-based fingerprinting is a technically efficient method in that it facilitates non-destructive analysis in a high-throughput manner. Furthermore, with the integrated analysis used here, we were able to discover unique metabotypes in rice-Arabidopsis FOX lines; thus, this approach is beneficial for investigating the function of rice genes related to metabolism.
Authors Suzuki, M., Kusano, M., Takahashi, H., Nakamura, Y., Hayashi, N., Kobayashi, M., Ichikawa, T., Matsui, M., Hirochika, H. and Saito, K.
Reference Metabolomics, March 2010, Volume 6, Issue 1, pp 137–145

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Analytical Method Details Information

Title GC-TOF/MS analysis
Instrument GC:Agilent 6890N gas chromatograph (Agilent Technologies, Wilmingston, USA)
MS:Pegasus III TOF mass spectrometer (LECO, St. Joseph, MI, USA)
Instrument Type
Ionization EI
Ion Mode Positive
Description To assess the metabolite profiles of the candidate lines that showed altered their metabolite fingerprints in FT-NIR analysis, 200 seeds of each of the candidate lines were extracted at a concentration of 10 mg/ml, derivatized, and then analyzed by GC-TOF/MS as described in Kusano et al (2007). A total of 266 metabolite peaks were extracted for each seed sample. Of them, 67 peaks were identified or annotated as known metabolites, 186 peaks were of unknown metabolites, and 13 peaks were annotated as mass spectral tags (MSTs) (Schauer et al. 2005).

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