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          Institute: MPI für molekulare Pflanzenphysiologie     Collection: Publikationen Pflanzenphysiologie     Display Documents



  history
ID: 228605.0, MPI für molekulare Pflanzenphysiologie / Publikationen Pflanzenphysiologie
Kinetics of labelling of organic and amino acids in potato tubers by gas chromatography-mass spectrometry following incubation in C-13 labelled isotopes
Authors:Roessner-Tunali, U.; Liu, J. L.; Leisse, A.; Balbo, I.; Perez-Melis, A.; Willmitzer, L.; Fernie, A. R.
Language:English
Date of Publication (YYYY-MM-DD):2004-08
Title of Journal:Plant Journal
Journal Abbrev.:Plant J
Volume:39
Issue / Number:4
Start Page:668
End Page:679
Review Status:not specified
Audience:Not Specified
Abstract / Description:Metabolic pathways of primary metabolism of discs isolated from potato tubers were evaluated by the use of a gas chromatography-mass spectrometry (GC-MS) method generated specifically for this purpose. After testing several possible methods including chemical ionization, it was decided for reasons of sensitivity, reproducibility and speed to use electron impact ionization-based GC-MS analysis. The specific labelling and label accumulation of over 30 metabolites including a broad number of sugars, organic and amino acids was analysed following the incubation of tuber discs in [U-C-13]glucose. The reproducibility of this method was similar to that found for other GC-MS-based analyses and comparison of flux estimates from this method with those obtained from parallel, yet less comprehensive, radiolabel experiments revealed close agreement. Therefore, the novel method allows quantitatively evaluation of a broad range of metabolic pathways without the need for laborious (and potentially inaccurate), chemical fractionation procedures commonly used in the estimation of fluxes following incubation in radiolabelled substrates. As a first experiment the GC-MS method has been applied to compare the metabolism of wild type and wel-characterized transgenic potato tubers exhibiting an enhanced sucrose mobilization. The fact that this method is able to rapidly yield further comprehensive information into primary metabolism illustrates its power as a further phenotyping tool for the analysis of plant metabolism.
Free Keywords:high-throughput metabolic analysis
; gas chromatography-mass spectrometry
; potato tuber
; stable isotope labelling
; low internal oxygen
; starch synthesis
; metabolic flux
; plastidial phosphoglucomutase
; sucrose phosphorylase
; solanum-tuberosum
; transgenic plants
; yeast invertase
; krebs cycle
; expression
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für molekulare Pflanzenphysiologie/Molekulare Physiologie/AG Willmitzer/Fernie
External Affiliations:Max Planck Inst Mol Pflanzenphysiol, Dept Mol Physiol, Muhlenberg 1, D-14476 Golm, Germany
Max Planck Inst Mol Pflanzenphysiol, Dept Mol Physiol, D-14476 Golm, Germany
Scottish Crop Res Inst, Computat Biol Programme, Dundee DD2 5DA, Scotland
Identifiers:ISI:000222944600014 [ID No:1]
ISI:000222944600014 [ID No:2]
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