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Lehmann, Marco M.; Egli, Melanie; Brinkmann, Nadine; Werner, Roland A.; Saurer, Matthias; Kahmen, Ansgar

Improving the extraction and purification of leaf and phloem sugars for oxygen isotope analyses

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  • Media type: E-Article
  • Title: Improving the extraction and purification of leaf and phloem sugars for oxygen isotope analyses
  • Contributor: Lehmann, Marco M.; Egli, Melanie; Brinkmann, Nadine; Werner, Roland A.; Saurer, Matthias; Kahmen, Ansgar
  • imprint: Wiley, 2020
  • Published in: Rapid Communications in Mass Spectrometry
  • Language: English
  • DOI: 10.1002/rcm.8854
  • ISSN: 0951-4198; 1097-0231
  • Origination:
  • Footnote:
  • Description: <jats:sec><jats:title>Rationale</jats:title><jats:p>The oxygen isotopic composition (here shown as the <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O value) of soluble sugars in leaves and phloem tissue holds valuable information about plant functions in response to climatic changes. However, <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O analysis of sugars is prone to error, and thoroughly tested methods are lacking.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>We performed three experiments to test if sample preparation modifies the <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values of sugars. In experiment 1, we tested the effects of oven‐drying versus freeze‐drying, whereas in experiment 2 we focused on the extraction and purification of leaf sugars. In experiment 3, we investigated the exudation and purification of twig phloem sugars as a function of exudation time and different ethylenediaminetetraacetic acid (EDTA) exudation media.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Freeze‐drying produced more consistent <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values than oven‐drying for sucrose but not for phloem sugars. The extraction and purification of leaf sugars can be performed without a significant modification of their <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values; yet the purified leaf and phloem sugars possessed higher <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values than the fraction of water‐soluble compounds. Moreover, the exudation time significantly modulated the <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values of phloem sugars, which is probably related to changes in the sugar composition. The addition of EDTA did not improve the determination of the <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values of phloem sugars.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>We show that the sample preparation of plant sugars for the reliable determination of <jats:italic>δ</jats:italic><jats:sup>18</jats:sup>O values requires a strict protocol, which is described in this paper. For phloem sugar, we recommend a maximum exudation time of 1 h to reduce the degradation of sucrose and minimise oxygen isotope exchange reactions between the resulting hexoses and water.</jats:p></jats:sec>