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Gas Chromatography–Mass Spectrometry-Based 13C-Labeling Studies in Plant Metabolomics

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Plant Metabolomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1778))

Abstract

Stable-isotope labeling analysis has been used to discover new metabolic pathways and their key regulatory points in a wide range of organisms. Given the complexity of the plant metabolic network, this analysis provides information complementary to that obtained from metabolite profiling that can be used to understand how plants cope with adverse conditions, and how metabolism varies between different cells, tissues, and organs. Here we describe the experimental procedures from sample harvesting and extraction to mass spectral analysis and interpretation that allow the researcher to perform 13C-labeling experiments. A wide range of plant material, from single cells to whole plants, can be used to investigate the metabolic fate of the 13C from a predefined tracer. Thus, a key point of this analysis is to choose the correct biological system, the substrate and the condition to be investigated; all of which implicitly relies on the biological question to be investigated. Rapid sample quenching and a careful data analysis are also critical points in such studies. By contrast to other metabolomic approaches, stable-isotope labeling can provide information concerning the fluxes through metabolic networks, which is essential for understanding and manipulating metabolic phenotypes and therefore of pivotal importance for both systems biology and plant metabolic engineering.

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Acknowledgments

Scholarships granted by the Coordination for the Improvement of Higher Level Personnel (CAPES) to V.F.L. and the National Council for Scientific and Technological Development (CNPq-Brazil) to L.P.S. are gratefully acknowledged. The Foundation for Support of Scientific and Technological Development of Ceará (FUNCAP) grant number AEP-0128-00092.01.00/17 to D.M.D. is gratefully acknowledged. 

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Authors and Affiliations

  1. Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Fortaleza, CE, Brazil

    Valéria F. Lima & Danilo M. Daloso

  2. Max Planck Institute of Molecular Plant Physiology, Potsdam-Golm, Germany

    Leonardo Perez de Souza & Alisdair R. Fernie

  3. Departamento de Botânica, Universidade de Brasília, Brasília, DF, Brazil

    Thomas C. R. Williams

Authors
  1. Valéria F. Lima
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  2. Leonardo Perez de Souza
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  3. Thomas C. R. Williams
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  4. Alisdair R. Fernie
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  5. Danilo M. Daloso
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Corresponding author

Correspondence to Danilo M. Daloso .

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Editors and Affiliations

  1. Plant Metabolomics Laboratory, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa (ITQB NOVA), Oeiras, Portugal

    Carla António

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Lima, V.F., de Souza, L.P., Williams, T.C.R., Fernie, A.R., Daloso, D.M. (2018). Gas Chromatography–Mass Spectrometry-Based 13C-Labeling Studies in Plant Metabolomics. In: António, C. (eds) Plant Metabolomics. Methods in Molecular Biology, vol 1778. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7819-9_4

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  • DOI: https://doi.org/10.1007/978-1-4939-7819-9_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7818-2

  • Online ISBN: 978-1-4939-7819-9

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