Towards improved quantification of internal diffusion resistances and CO2 isotope exchange in leaves

The oxygen isotopic composition (δ18O) of atmospheric CO2 can constrain the C cycle at a range of scales, in particular partitioning net CO2 exchanges into their component gross fluxes and providing insights to linked C and water fluxes. There are significant limitations to utilizing the δ18O of CO2 in this way, however, because of uncertainties associated with the isotopic exchange of CO2 with terrestrial water pools. Leaf water represents a critical pool with ongoing debates about its enrichment in heavy isotopes during transpiration and the hydration of CO2 and the resulting oxygen isotope exchange. Isotopic heterogeneity of the leaf water, the spatial distribution and activity of carbonic anhydrase (CA) within leaves, and resistance to diffusion of CO2 from the substomatal cavity to chloroplasts are all key components with important uncertainties. Better constraints on these would significantly improve our ability to understand and model the global C budget as well as yield insights to fundamental aspects of leaf physiology. We report results using a new measurement system that permits the simultaneous measurement of the 13C and 18O composition of CO2 and the 18O isotopic composition of leaf transpiration. As this new approach permits rapid alteration of the isotopic composition of gases interacting with the leaf, key model parameters can be derived directly and simultaneously. Hence, our approach dos not rely on separate measurements shifted in time from the gas exchange measurements or that may not quantify the relevant scale of heterogeneity (e.g., CA enzyme assays or bulk leaf water extraction and analysis). In particular, this new method explicitly distinguishes the leaf mesophyll resistance to CO2 transport relevant for photosynthesis from the resistance required for interpreting the d18O of CO2. This new measurement system and modeling approach allows a thorough assessment of key elements of our current understanding of CO2-H2O oxygen isotope exchange in terrestrial ecosystems.