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High-Throughput Extraction and Enzymatic Determination of Sugars and Fructans in Fructan-Accumulating Plants

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Plant Circadian Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2398))

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Abstract

Fructans are carbohydrates present in more than 15% of flowering plants. They represent the major pool of carbohydrates in some species, especially when facing cold or drought. However, the functions of fructans with high or low degrees of polymerization (DP), their diurnal use, and the regulation of their synthesis and degradation in response to stresses still remain unclear. Here we present an enzymatic protocol adapted to 96-well microplates that simultaneously allows the determination of fructans and glucose, fructose, and sucrose. Moreover, the protocol allows to estimate the average DP of the fructans in the samples. The protocol is based on the enzymatic degradation of fructans into glucose and fructose and their subsequent conversion into gluconate 6-phosphate concomitant with the formation of NADH in the presence of ATP.

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Acknowledgments

Kallyne Barros was supported by a postgraduate studentship funded by CNPQ (ID. 233005/2014-5) and Auxiliadora Oliveira Martins by CNPq (ID. 210299/2015-0). Ronan Sulpice and Masami Inaba were supported by a Research Stimulus Grant (VICCI—Grant No: 14/S/81) funded by the Irish Department of Agriculture, Food and the Marine (DAFM).

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Authors and Affiliations

  1. NUI Galway, Plant Systems Biology Laboratory, Ryan Institute, School of Natural Sciences, Galway, Ireland

    Kallyne A. Barros, Masami Inaba, Auxiliadora Oliveira Martins & Ronan Sulpice

  2. Departamento de Biologia Vegetal, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil

    Auxiliadora Oliveira Martins

Authors
  1. Kallyne A. Barros
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  2. Masami Inaba
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  4. Ronan Sulpice
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Correspondence to Ronan Sulpice .

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Editors and Affiliations

  1. Department of Biology, University of Bielefeld, Bielefeld, Nordrhein-Westfalen, Germany

    Dorothee Staiger

  2. Department of Biology, University of York, Heslington, UK

    Seth Davis

  3. Department of Biology, University of York, Heslington, UK

    Amanda Melaragno Davis

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Barros, K.A., Inaba, M., Martins, A.O., Sulpice, R. (2022). High-Throughput Extraction and Enzymatic Determination of Sugars and Fructans in Fructan-Accumulating Plants. In: Staiger, D., Davis, S., Davis, A.M. (eds) Plant Circadian Networks. Methods in Molecular Biology, vol 2398. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1912-4_10

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  • DOI: https://doi.org/10.1007/978-1-0716-1912-4_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1911-7

  • Online ISBN: 978-1-0716-1912-4

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