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Intra- and extracellular carbohydrates in plant cell cultures investigated by 1H-NMR

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Abstract

With the aim of quantifying intra- and extracellular carbohydrates media and cell-extracts from a Tabernaemontana divaricata plant cell-suspension culture were investigated with 1H-NMR.

For suppression of the solvent peak the Meiboom-Gill modification of the Carr-Purcell (CPMG) spin-echo sequence was used after addition of a paramagnetic relaxation agent (Mn2+) to the sample. Several aspects of this method were optimized (the manganese concentration, the interpulse delay and the number of spin-echo cycles) so as to obtain a rapid and easy method in which no pretreatment of media or cell-extracts was needed. Besides the speed and ease of the method, also the direct identification of carbohydrates and other main components is an advantage.

The exhaustion of extracellular carbohydrates was found to coincide with the maximum amount of intracellular carbohydrates. The intracellular carbohydrates, i.e. glucose and fructose, were consumed at a low rate, during several weeks.

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Abbreviations

1H-NMR:

proton nuclear magnetic resonance

2,4-D:

2,4-dichlorophenoxyacetic acid

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Authors and Affiliations

  1. Division of Pharmacognosy, BDL Project group Plant Cell Biotechnology, Center for Bio-Pharmaceutical Sciences, Leiden University, Gorlaeus Laboratories, P.O. Box 9502, 2300, RA Leiden, The Netherlands

    J. Schripsema & R. Verpoorte

  2. NMR division, Department of Chemistry, Leiden University, Gorlaeus Laboratories, P.O. Box 9502, 2300, RA Leiden, The Netherlands

    C. Erkelens

Authors
  1. J. Schripsema
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  2. C. Erkelens
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  3. R. Verpoorte
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Communicated by W. Barz

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Schripsema, J., Erkelens, C. & Verpoorte, R. Intra- and extracellular carbohydrates in plant cell cultures investigated by 1H-NMR. Plant Cell Reports 9, 527–530 (1991). https://doi.org/10.1007/BF00232112

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  • DOI: https://doi.org/10.1007/BF00232112

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