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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.


Metabolic map of pennycress embryos at different stages of development. Values are expressed in pmol per embryo and are the average ±SD of three biological replicates from embryos harvested at 11, 13, 15, 17, 19, and 21 DAP. SUC, sucrose; FRU, fructose; GLC, glucose; INO, inositol; GLY, glycerol; Ery/Thr, erythritol/threitol; Ala, alanine; Arg, arginine; Asn, asparagine; Asp, aspartate; Cys, cysteine; Lys, lysine; Gln, glutamine; Glu, glutamate; Gly, glycine; His, histidine; OHPro, hydroxyproline; Leu, leucine; Ile, isoleucine; Met, methionine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; GABA, 4-aminobutyric acid; Orn, ornithine; Citru, citrulline; T6P, trehalose 6-phosphate; UDPG, UDP-glucose; SUCP, sucrose 6-phosphate; G1P, glucose 1-phosphate; M1P/G1P, mannose 1-phosphate/glucose 1-phosphate; F6P, fructose 6-phosphate; G6P, glucose 6-phosphate; 6PG, 6-phosphogluconic acid; P5P, pentose 5-phosphate; R1,5-bP, ribulose 1,5-bisphosphate; S7P, sedoheptulose 7-phosphate; E4P, eryhtrose 4-phosphate; F1,6bP, fructose 1,6-bisphosphate; GLYP, glycerol-phosphates; TP, triose phosphates; PGA, 2–3 phosphoglycerates; dX5P, deoxyxylulose 5-phosphate; PEP, phosphoenolpyruvate; SHI, shikimate; PYR, pyruvate; AcCoA, acetyl-CoA; CIT, citrate; cisACO, cis-aconitate; isoCIT, isocitrate; AKG, α-ketoglutarate; SUCC, succinate; FUM, fumarate; MAL, malate; OAA, oxaloacetate. Metabolites coloured in red and green correspond to glycolysis and the TCA cycle, respectively.
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Figure 3: Metabolic map of pennycress embryos at different stages of development. Values are expressed in pmol per embryo and are the average ±SD of three biological replicates from embryos harvested at 11, 13, 15, 17, 19, and 21 DAP. SUC, sucrose; FRU, fructose; GLC, glucose; INO, inositol; GLY, glycerol; Ery/Thr, erythritol/threitol; Ala, alanine; Arg, arginine; Asn, asparagine; Asp, aspartate; Cys, cysteine; Lys, lysine; Gln, glutamine; Glu, glutamate; Gly, glycine; His, histidine; OHPro, hydroxyproline; Leu, leucine; Ile, isoleucine; Met, methionine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; GABA, 4-aminobutyric acid; Orn, ornithine; Citru, citrulline; T6P, trehalose 6-phosphate; UDPG, UDP-glucose; SUCP, sucrose 6-phosphate; G1P, glucose 1-phosphate; M1P/G1P, mannose 1-phosphate/glucose 1-phosphate; F6P, fructose 6-phosphate; G6P, glucose 6-phosphate; 6PG, 6-phosphogluconic acid; P5P, pentose 5-phosphate; R1,5-bP, ribulose 1,5-bisphosphate; S7P, sedoheptulose 7-phosphate; E4P, eryhtrose 4-phosphate; F1,6bP, fructose 1,6-bisphosphate; GLYP, glycerol-phosphates; TP, triose phosphates; PGA, 2–3 phosphoglycerates; dX5P, deoxyxylulose 5-phosphate; PEP, phosphoenolpyruvate; SHI, shikimate; PYR, pyruvate; AcCoA, acetyl-CoA; CIT, citrate; cisACO, cis-aconitate; isoCIT, isocitrate; AKG, α-ketoglutarate; SUCC, succinate; FUM, fumarate; MAL, malate; OAA, oxaloacetate. Metabolites coloured in red and green correspond to glycolysis and the TCA cycle, respectively.

Mentions: For the purpose of quantifying the compounds characterized by metabolite profiling, boiling water extraction was performed on pennycress embryos harvested at different stages of development. Extracted compounds were then analysed by LC-MS/MS and quantified according to 13C-labelled internal standards as well as standard curves generated for each metabolite. The percentage recovery of this method was previously determined in plant tissues for each metabolite (Cocuron et al., 2014). Intermediates from glycolysis, the OPPP, the TCA cycle, and the Calvin cycle were measured by LC-MS/MS, indicating that all these pathways are active in developing pennycress embryos (Fig. 3; Supplementary Table S2 at JXB online).


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)

Metabolic map of pennycress embryos at different stages of development. Values are expressed in pmol per embryo and are the average ±SD of three biological replicates from embryos harvested at 11, 13, 15, 17, 19, and 21 DAP. SUC, sucrose; FRU, fructose; GLC, glucose; INO, inositol; GLY, glycerol; Ery/Thr, erythritol/threitol; Ala, alanine; Arg, arginine; Asn, asparagine; Asp, aspartate; Cys, cysteine; Lys, lysine; Gln, glutamine; Glu, glutamate; Gly, glycine; His, histidine; OHPro, hydroxyproline; Leu, leucine; Ile, isoleucine; Met, methionine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; GABA, 4-aminobutyric acid; Orn, ornithine; Citru, citrulline; T6P, trehalose 6-phosphate; UDPG, UDP-glucose; SUCP, sucrose 6-phosphate; G1P, glucose 1-phosphate; M1P/G1P, mannose 1-phosphate/glucose 1-phosphate; F6P, fructose 6-phosphate; G6P, glucose 6-phosphate; 6PG, 6-phosphogluconic acid; P5P, pentose 5-phosphate; R1,5-bP, ribulose 1,5-bisphosphate; S7P, sedoheptulose 7-phosphate; E4P, eryhtrose 4-phosphate; F1,6bP, fructose 1,6-bisphosphate; GLYP, glycerol-phosphates; TP, triose phosphates; PGA, 2–3 phosphoglycerates; dX5P, deoxyxylulose 5-phosphate; PEP, phosphoenolpyruvate; SHI, shikimate; PYR, pyruvate; AcCoA, acetyl-CoA; CIT, citrate; cisACO, cis-aconitate; isoCIT, isocitrate; AKG, α-ketoglutarate; SUCC, succinate; FUM, fumarate; MAL, malate; OAA, oxaloacetate. Metabolites coloured in red and green correspond to glycolysis and the TCA cycle, respectively.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 3: Metabolic map of pennycress embryos at different stages of development. Values are expressed in pmol per embryo and are the average ±SD of three biological replicates from embryos harvested at 11, 13, 15, 17, 19, and 21 DAP. SUC, sucrose; FRU, fructose; GLC, glucose; INO, inositol; GLY, glycerol; Ery/Thr, erythritol/threitol; Ala, alanine; Arg, arginine; Asn, asparagine; Asp, aspartate; Cys, cysteine; Lys, lysine; Gln, glutamine; Glu, glutamate; Gly, glycine; His, histidine; OHPro, hydroxyproline; Leu, leucine; Ile, isoleucine; Met, methionine; Phe, phenylalanine; Pro, proline; Ser, serine; Thr, threonine; Tyr, tyrosine; Val, valine; GABA, 4-aminobutyric acid; Orn, ornithine; Citru, citrulline; T6P, trehalose 6-phosphate; UDPG, UDP-glucose; SUCP, sucrose 6-phosphate; G1P, glucose 1-phosphate; M1P/G1P, mannose 1-phosphate/glucose 1-phosphate; F6P, fructose 6-phosphate; G6P, glucose 6-phosphate; 6PG, 6-phosphogluconic acid; P5P, pentose 5-phosphate; R1,5-bP, ribulose 1,5-bisphosphate; S7P, sedoheptulose 7-phosphate; E4P, eryhtrose 4-phosphate; F1,6bP, fructose 1,6-bisphosphate; GLYP, glycerol-phosphates; TP, triose phosphates; PGA, 2–3 phosphoglycerates; dX5P, deoxyxylulose 5-phosphate; PEP, phosphoenolpyruvate; SHI, shikimate; PYR, pyruvate; AcCoA, acetyl-CoA; CIT, citrate; cisACO, cis-aconitate; isoCIT, isocitrate; AKG, α-ketoglutarate; SUCC, succinate; FUM, fumarate; MAL, malate; OAA, oxaloacetate. Metabolites coloured in red and green correspond to glycolysis and the TCA cycle, respectively.
Mentions: For the purpose of quantifying the compounds characterized by metabolite profiling, boiling water extraction was performed on pennycress embryos harvested at different stages of development. Extracted compounds were then analysed by LC-MS/MS and quantified according to 13C-labelled internal standards as well as standard curves generated for each metabolite. The percentage recovery of this method was previously determined in plant tissues for each metabolite (Cocuron et al., 2014). Intermediates from glycolysis, the OPPP, the TCA cycle, and the Calvin cycle were measured by LC-MS/MS, indicating that all these pathways are active in developing pennycress embryos (Fig. 3; Supplementary Table S2 at JXB online).

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.