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Coupling Radiotracer Experiments with Chemical Fractionation for the Estimation of Respiratory Fluxes

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Plant Respiration and Internal Oxygen

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1670))

Abstract

Carbohydrates catabolized via respiratory processes are not only used for energy production but also for biosynthesis of cellular components including soluble molecules (sugars, amino acids, organic acids, and their derivatives) and insoluble macromolecules (proteins, starch, and cell wall). Radiotracer experiments using 14C-labeled glucose provide a global picture of the fate of respired carbon in the metabolic network. This method is based on a chemical fractionation of biomolecules in 14C-glucose fed plant materials and the subsequent determination of radioactivity in each fraction. Metabolic flux into each fraction can be estimated from the specific activity of the hexose phosphate pool. Here, we describe the procedure for glucose metabolism in potato tuber but similar protocols can be adopted for various plant organs and substrates.

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Acknowledgment

This study was supported by DFG (OB438 to T.O.), the European Commission’s Directorate General for Research within the 7th Framework Program (FP7/2007-2013 grant no. 270089 [MULTIBIOPRO] to T.O. and A.R.F.), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq to L.R.S), and Max-Planck Society (to A.R.F.). We thank Saleh Alseekh for useful comments.

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Authors and Affiliations

  1. Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Potsdam-Golm, Germany

    Toshihiro Obata, Laise Rosado-Souza & Alisdair R. Fernie

  2. Department of Biochemistry, University of Nebraska Lincoln, 1901 Vine St, 68588, Lincoln, NE, USA

    Toshihiro Obata

Authors
  1. Toshihiro Obata
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  2. Laise Rosado-Souza
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  3. Alisdair R. Fernie
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Corresponding author

Correspondence to Alisdair R. Fernie .

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Editors and Affiliations

  1. National Institute of Plant Genome Research, New Delhi, Delhi, India

    Kapuganti Jagadis Gupta

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Obata, T., Rosado-Souza, L., Fernie, A.R. (2017). Coupling Radiotracer Experiments with Chemical Fractionation for the Estimation of Respiratory Fluxes. In: Jagadis Gupta, K. (eds) Plant Respiration and Internal Oxygen. Methods in Molecular Biology, vol 1670. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7292-0_2

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  • DOI: https://doi.org/10.1007/978-1-4939-7292-0_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7291-3

  • Online ISBN: 978-1-4939-7292-0

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