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Understanding leaf wax regeneration and recycling is crucial for plant physiology and paleoclimate studies. However, our recent isotope labeling experiments on a grass species (Phleum pratense) yielded different conclusions from published data on a tree species (Populus trichocarpa), with the former showing rapid regeneration and the latter little regeneration in mature leaves. It is therefore important to determine if the discrepancies in published results were due to differing dynamics of leaf wax regeneration and/or caveats in experimental methods.

Leaves from a native New England tree species (Fraxinus americana) were collected at 1 to 3 h intervals over a 2‐day experimental period, and, subsequently, the leaf wax δ²H isotopic ratios were measured using gas chromatography/isotope ratio mass spectrometry.

It was necessary to irrigate the tree using water with …