Abstract

ABSTRACTTwo direct but independent approaches were developed to identify the average δ18O value of the water fraction in the chloroplasts of transpiring leaves. In the first approach, we used the δ18O value of CO2 in isotopic equilibrium with leaf water to reconstruct the δ18O value of water in the chloroplasts. This method was based on the idea that the enzyme carbonic anhydrase facilitates isotopic equilibrium between CO2 and H2O predominantly in the chloroplasts, at a rate that is several orders of magnitude faster than the non‐catalysed exchange in other leaf water fractions. In the second approach, we measured the δ18O value of O2 from photosynthetic water oxidation in the chloroplasts of intact leaves. Since O2 is produced from chloroplast water irreversibly and without discrimination, the δ18O value of the O2 should be identical to that of chloroplast water. In intact, transpiring leaves of sunflower (Helianthus annuus cv. giant mammoth) under the experimental conditions used, the average δ18O value of chloroplasts water was displaced by 3—10 % (depending on relative humidity and atmospheric composition) below the value predicted by the conventional Craig & Gordon model. Furthermore, this δ18O value was always lower than the δ18O value that was measured for bulk leaf water. Our results have implications for a variety of environmental studies since it is the δ18O value of water in the chloroplasts that is the relevant quantity in considering terrestrial plants influence on the δ18O values of atmospheric CO2 and O2, as well as in influencing the δ18O of plant organic matter.

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