Rationale
The oxygen isotopic composition (here shown as the δ18O value) of soluble sugars in leaves and phloem tissue holds valuable information about plant functions in response to climatic changes. However, δ18O analysis of sugars is prone to error, and thoroughly tested methods are lacking.
Methods
We performed three experiments to test if sample preparation modifies the δ18O 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.
Results
Freeze‐drying produced more consistent δ18O 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 δ18O values; yet the purified leaf and phloem sugars possessed higher δ18O values than the fraction of water‐soluble compounds. Moreover, the exudation time significantly modulated the δ18O 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 δ18O values of phloem sugars.
Conclusions
We show that the sample preparation of plant sugars for the reliable determination of δ18O 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.