Effects of grass leaf anatomy, development and light/dark alternation on the triple oxygen isotope signature of leaf water and phytoliths: insights for a new proxy of continental atmospheric humidity

Abstract. Continental relative humidity (RH) is a key-climate parameter. However, there is a lack of quantitative RH proxies suitable for climate model-data comparisons. Recently, a combination of climate chamber and natural transect calibrations laid the groundwork for examining the robustness of the triple oxygen isotope composition (δ18O, δ17O) of phytoliths as a new proxy for past changes in RH. However, it was recommended that besides RH, additional factors that may impact δ18O and δ17O of plant water and phytoliths be examined. Here, the effects of leaf anatomy, leaf development stage and day/night alternations are addressed from the growth of the grass species F. arundinacea in climate chambers. Plant water and phytoliths are analyzed in δ18O and δ17O. Silicification patterns are examined using light and scanning electron observation of phytoliths. The isotope data show the increasing contribution of evaporated epidermal water to the bulk leaf water, from sheath to proximal and apical leaf blade. However, despite this isotope heterogeneity, δ18O and δ17O of the bulk leaf water can be predicted by the Craig and Gordon model, in the given experimental conditions (high RH). Regarding phytoliths, their forming water (mainly epidermal) is, as expected, more impacted by evaporation than the bulk leaf water. This discrepancy increases from sheath to proximal and apical blade and can be explained by the steepening of the radial concentration gradient of evaporated water along the leaf. However, we show that because most of silica polymerizes in epidermal long cells of the apical blade of the leaves, the δ18O and δ17O of bulk grass phytoliths should not be impacted by the diversity in grass anatomy. The data additionally show that most of silica polymerizes at the end of the leaf elongation stage and at the transition towards leaf senescence. Thus, climate conditions at that time should be considered when interpreting δ18O and δ17O of phytoliths from the natural environment. At least, no light/dark effect was detected on the δ18O and δ17O signature of plant water and phytoliths of F. arundinacea. However, when day/night alternations are characterized by significant changes in RH, the lowest RH conditions favoring evaporation and silica polymerization should be considered when calibrating the phytolith proxy. This study contributes to the identification of the parameters driving the δ18O and δ17O of bulk grass phytoliths. It additionally brings elements to further understand and model the δ18O and δ17O of grass leaf water, which influences the isotope signal of several processes at the soil/plant/atmosphere interface.