Comparative cryogenic extraction rehydration experiments reveal isotope fractionation during root water uptake in...

Determining whether isotope fractionation occurs during root water uptake is a prerequisite for using stem or xylem water isotopes to trace water sources. However, it is unclear whether isotope fractionation occurs during root water uptake in gramineous crops. We conducted prevalidation experiments to estimate the isotope measurement bias associated with cryogenic vacuum distillation (CVD). Next, we assessed isotope fractionation during root water uptake in two common agronomic crops, wheat (Triticum aestivum L.) and maize (Zea mays L.), under flooding after postdrought stress conditions. Cryogenic vacuum distillation caused significant depletion of ² H but negligible effects on ¹⁸ O for both soil and stem water. Surprisingly CVD caused depletion of ² H and enrichment of ¹⁸ O in root water. Stem and root water δ¹⁸ O were more than soil water δ¹⁸ O, even considering the uncertainty of CVD. Soil water ¹⁸ O was depleted compared with irrigation water ¹⁸ O in the pots with plants but enriched relative to irrigation water ¹⁸ O in the pots without plants. These results indicate that isotope fractionation occurred during wheat and maize root water uptake after full irrigation and led to a heavy isotope enrichment in stem water. Therefore, the xylem/stem water isotope approach widely used to trace water sources should be carefully evaluated.