Comparing plant water extraction methods for isotopic analyses: the Scholander pressure chamber vs. the cryogenic vacuum distillation

Recent studies applying stable isotopes of hydrogen and oxygen showed that different methods for extracting water from plant tissues can return different isotopic composition. One of the most used methods to extract plant water is the cryogenic vacuum distillation (CVD), which tends to extract total plant water. Conversely, the Scholander pressure chamber (SPC), which is commonly used by tree physiologists to measure water potential in plant tissues, likely accesses only the mobile plant water (i.e., xylem and inter-cellular water). However, only few studies reported the application of SPC to extract plant water for isotopic analyses, and therefore, an inter-method comparison between SPC and CVD is needed.In this study, we analyzed the variability in the isotopic signature of plant water extracted by SPC and CVD. Furthermore, we considered the potential variability in the isotopic composition of the plant water extracted from various tissues by CVD (i.e., leaves, twig without bark, twig with bark, twig close to the trunk of the tree, and wood core), and from different tree species (i.e., alder, apple, chestnut and beech) located in three different study areas in northern Italy.Results indicate that plant waters extracted by SPC and CVD were significantly different, likely due to the extraction of different plant water domains. The difference in the isotopic composition obtained by the two extraction methods was smaller in the beech samples compared to alder, apple and chestnut samples. The signature of alder, apple and chestnut plant water extracted by SPC was more enriched in heavy isotopes than the samples obtained by CVD (except for the leaf water obtained by CVD, which also had a marked evaporative signature). We conclude that plant water extraction by SPC does not represent an alternative for CVD, as SPC likely extracts mostly the mobile plant water, whereas CVD tends to retrieve all water stored in the sampled tissues. However, studies aiming to quantify the relative contribution of the water sources to transpiration should rely more on the isotopic composition of xylem water transpiring during the sampling day (theoretically sampled by SPC), than the isotopic composition of total plant water (sampled by CVD), which also contains a fraction of water that could be stored in plant tissues for a long time. Keywords: stable isotopes of hydrogen and oxygen; cryogenic vacuum distillation; Scholander pressure chamber; plant water; xylem water.