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Metabolite fingerprinting of pennycress (Thlaspi arvense L.) embryos to assess active pathways during oil synthesis.

Tsogtbaatar E, Cocuron JC, Sonera MC, Alonso AP - J. Exp. Bot. (2015)

Bottom Line: Secondly, these intermediates were quantified in developing pennycress embryos by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring mode.Finally, partitional clustering analysis grouped the intracellular metabolites that shared a similar pattern of accumulation over time into eight clusters.This study underlined that: (i) sucrose might be stored rather than cleaved into hexoses; (ii) glucose and glutamine would be the main sources of carbon and nitrogen, respectively; and (iii) glycolysis, the oxidative pentose phosphate pathway, the tricarboxylic acid cycle, and the Calvin cycle were active in developing pennycress embryos.

View Article: PubMed Central - PubMed

Affiliation: The Ohio State University, Department of Molecular Genetics, Columbus, OH 43210, USA.

No MeSH data available.


Metabolite profiling of pennycress embryos at 17 DAP. (A) GC-MS chromatogram of 17 DAP pennycress embryos obtained after MSTFA derivatization. Enlarged chromatogram areas depicting the main classes of compounds, (B) amino acids, (C) organic acids, and (D) sugars found in pennycress embryos. The NIST 11 library was used to assign the different peaks.
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Figure 2: Metabolite profiling of pennycress embryos at 17 DAP. (A) GC-MS chromatogram of 17 DAP pennycress embryos obtained after MSTFA derivatization. Enlarged chromatogram areas depicting the main classes of compounds, (B) amino acids, (C) organic acids, and (D) sugars found in pennycress embryos. The NIST 11 library was used to assign the different peaks.

Mentions: During the developmental process, embryos produce a wide variety of metabolites in a temporal fashion as a result of changes in their metabolism. Metabolite profiling, also known as untargeted metabolomics, enables the detection of intracellular compounds at a given time. Through this approach one can gain qualitative (rather than quantitative) information about specific classes of intermediates accumulating at the same time as the synthesis of a product of interest. In this study, metabolite profiling was used to characterize all the compounds that were present during fatty acid synthesis. For this purpose, intracellular metabolites were extracted from 17 DAP pennycress embryos with cold methanol:chloroform:water (MCW 2.5:1:1, v:v:v) (Fiehn, 2006) or boiling water (Alonso et al., 2010b) and then were chemically modified with MSTFA+1% TMCS (Koek et al., 2006). Through the comparison between GC-MS profiles of the derivatized metabolites, boiling water was shown to be the most suitable method, enabling the detection of 385 peaks versus 344 for MCW (data not shown). A total of 112 peaks out of 385 were assigned with a probability ≥50% using the NIST 11 library (Fig. 2; Supplementary Table S1 at JXB online). The identification of the detected peaks qualitatively showed the presence of three main classes of metabolites (sugars, amino acids, and organic acids), and to a lesser extent, alkaloids, polyamines, phosphorylated metabolites, and free fatty acids (Fig. 2; Supplementary Table S1).


Metabolite fingerprinting of pennycress (Thlaspi arvense L.) embryos to assess active pathways during oil synthesis.

Tsogtbaatar E, Cocuron JC, Sonera MC, Alonso AP - J. Exp. Bot. (2015)

Metabolite profiling of pennycress embryos at 17 DAP. (A) GC-MS chromatogram of 17 DAP pennycress embryos obtained after MSTFA derivatization. Enlarged chromatogram areas depicting the main classes of compounds, (B) amino acids, (C) organic acids, and (D) sugars found in pennycress embryos. The NIST 11 library was used to assign the different peaks.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493779&req=5

Figure 2: Metabolite profiling of pennycress embryos at 17 DAP. (A) GC-MS chromatogram of 17 DAP pennycress embryos obtained after MSTFA derivatization. Enlarged chromatogram areas depicting the main classes of compounds, (B) amino acids, (C) organic acids, and (D) sugars found in pennycress embryos. The NIST 11 library was used to assign the different peaks.
Mentions: During the developmental process, embryos produce a wide variety of metabolites in a temporal fashion as a result of changes in their metabolism. Metabolite profiling, also known as untargeted metabolomics, enables the detection of intracellular compounds at a given time. Through this approach one can gain qualitative (rather than quantitative) information about specific classes of intermediates accumulating at the same time as the synthesis of a product of interest. In this study, metabolite profiling was used to characterize all the compounds that were present during fatty acid synthesis. For this purpose, intracellular metabolites were extracted from 17 DAP pennycress embryos with cold methanol:chloroform:water (MCW 2.5:1:1, v:v:v) (Fiehn, 2006) or boiling water (Alonso et al., 2010b) and then were chemically modified with MSTFA+1% TMCS (Koek et al., 2006). Through the comparison between GC-MS profiles of the derivatized metabolites, boiling water was shown to be the most suitable method, enabling the detection of 385 peaks versus 344 for MCW (data not shown). A total of 112 peaks out of 385 were assigned with a probability ≥50% using the NIST 11 library (Fig. 2; Supplementary Table S1 at JXB online). The identification of the detected peaks qualitatively showed the presence of three main classes of metabolites (sugars, amino acids, and organic acids), and to a lesser extent, alkaloids, polyamines, phosphorylated metabolites, and free fatty acids (Fig. 2; Supplementary Table S1).

Bottom Line: Secondly, these intermediates were quantified in developing pennycress embryos by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in multiple reaction monitoring mode.Finally, partitional clustering analysis grouped the intracellular metabolites that shared a similar pattern of accumulation over time into eight clusters.This study underlined that: (i) sucrose might be stored rather than cleaved into hexoses; (ii) glucose and glutamine would be the main sources of carbon and nitrogen, respectively; and (iii) glycolysis, the oxidative pentose phosphate pathway, the tricarboxylic acid cycle, and the Calvin cycle were active in developing pennycress embryos.

View Article: PubMed Central - PubMed

Affiliation: The Ohio State University, Department of Molecular Genetics, Columbus, OH 43210, USA.

No MeSH data available.