Field-based assessment of transpiration phenology in boreal tree species is a significant challenge. Here we develop an objective approach that uses stem radius change and its correlation with sapwood temperature to determine the timing of phenological changes in transpiration in mixed evergreen species. We test the stem-temp approach using a five year stem-radius dataset from black spruce (Picea mariana) and jack pine (Pinus banksiana) trees in Saskatchewan (2016–2020). We further compare transpiration phenological transition dates from this approach with tower-based phenological assessment from green chromatic coordinate derived from phenocam images, eddy-covariance-derived evapotranspiration and carbon uptake, tower-based measurements of solar-induced chlorophyll fluorescence and snowmelt timing. The stem-temp approach identified the start and end of four key transpiration phenological phases: (i) the end of temperature-driven cycles indicating the start of biological activity, (ii) the onset of stem rehydration, (iii) the onset of transpiration, and (iv) the end of transpiration-driven cycles. The proposed method is thus useful for characterizing the timing of changes in transpiration phenology and provides information about distinct processes that cannot be assessed with canopy-level phenological measurements alone.

Studies of tree water source partitioning have primarily focused on the growing season. However, little is yet known about the source of transpiration before, during, and after snowmelt when trees rehydrate and recommence transpiration in the spring. This study investigates tree water use during spring snowmelt following tree's winter stem shrinkage. We document the source of transpiration of three boreal forest tree species—Pinus banksiana, Picea mariana, and Larix laricina—by combining observations of weekly isotopic signatures (δ18O and δ2H) of xylem, soil water, rainfall and snowmelt with measurements of soil moisture dynamics, snow depth and high-resolution temporal measurements of stem radius changes and sap flow. Our data shows that the onset of stem rehydration and transpiration overlaps with snowmelt for evergreens. During rehydration and transpiration onset, xylem water at the canopy reflected a constant pre-melt isotopic signature likely showing late fall conditions. As snowmelt infiltrates the soil and recharges the soil matrix, soil water shows a rapid isotopic shift to depleted-snowmelt water values. While there was an overlap between snowmelt and transpiration timing, xylem and soil water isotopic values did not overlap during transpiration onset. Our data showed 1–2-week delay in the shift in xylem water from pre-melt to clear snowmelt-depleted water signatures in evergreen species. This delay appears to be controlled by tree water transit time that was in the order of 9–18 days. Our study shows that snowmelt is a key source for stem rehydration and transpiration in the boreal forest during spring onset.

Studies of tree water source partitioning have primarily focused on the growing season. However, little is yet known about the source of transpiration before, during, and after snowmelt when trees rehydrate and recommence transpiration in the spring. This study investigates tree water use during spring snowmelt following tree's winter stem shrinkage. We document the source of transpiration of three boreal forest tree species—Pinus banksiana, Picea mariana, and Larix laricina—by combining observations of weekly isotopic signatures (δ18O and δ2H) of xylem, soil water, rainfall and snowmelt with measurements of soil moisture dynamics, snow depth and high-resolution temporal measurements of stem radius changes and sap flow. Our data shows that the onset of stem rehydration and transpiration overlaps with snowmelt for evergreens. During rehydration and transpiration onset, xylem water at the canopy reflected a constant pre-melt isotopic signature likely showing late fall conditions. As snowmelt infiltrates the soil and recharges the soil matrix, soil water shows a rapid isotopic shift to depleted-snowmelt water values. While there was an overlap between snowmelt and transpiration timing, xylem and soil water isotopic values did not overlap during transpiration onset. Our data showed 1–2-week delay in the shift in xylem water from pre-melt to clear snowmelt-depleted water signatures in evergreen species. This delay appears to be controlled by tree water transit time that was in the order of 9–18 days. Our study shows that snowmelt is a key source for stem rehydration and transpiration in the boreal forest during spring onset.

Abstract The stable isotopes of hydrogen and oxygen in xylem water are often used to investigate tree water sources. But this traditional approach does not acknowledge the contribution of water stored in the phloem to transpiration and how this may affect xylem water and source water interpretations. Additionally, there is a prevailing assumption that there is no isotope fractionation during tree water transport. Here, we systematically sampled xylem and phloem water at daily and subdaily resolutions in a large lysimeter planted with Salix viminalis . Stem diurnal change in phloem water storage and transpiration rates were also measured. Our results show that phloem water is significantly less enriched in heavy isotopes than xylem water. At subdaily resolution, we observed a larger isotopic difference between xylem and phloem during phloem water refilling and under periods of tree water deficit. These findings contrast with the expectation of heavy‐isotope enriched water in phloem due to downward transport of enriched leaf water isotopic signatures. Because of previous evidence of aquaporin mediated phloem and xylem water transport and higher osmotic permeability of lighter hydrogen isotopologues across aquaporins, we propose that radial water transport across the xylem–phloem boundary may drive the relative depletion of heavy isotopes in phloem and their relative enrichment in xylem.

Abstract Ecohydrological investigations commonly use the stable isotopes of water (hydrogen and oxygen) as conservative ecosystem tracers. This approach requires accessing and analysing water from plant and soil matrices. Generally, there are six steps involved to retrieve hydrogen and oxygen isotope values from these matrices: (1) sampling, (2) sample storage and transport, (3) extraction, (4) pre‐analysis processing, (5) isotopic analysis, and (6) post‐processing and correction. At each step, cumulative errors can be introduced which sum to non‐trivial magnitudes. These can impact subsequent interpretations about water cycling and partitioning through the soil–plant‐atmosphere continuum. At each of these steps, there are multiple possible options to select from resulting in tens of thousands of possible combinations used by researchers to go from plant and soil samples to isotopic data. In a newly emerging field, so many options can create interpretive confusion and major issues with data comparability. This points to the need for development of shared standardized approaches. Here we critically examine the state of the process chain, reflecting on the issues associated with each step, and provide suggestions to move our community towards standardization. Assessing this shared ‘process chain’ will help us see the problem in its entirety and facilitate community action towards agreed upon standardized approaches.

Source water apportionment studies using the dual isotopes of oxygen and hydrogen have revolutionized our understanding of ecohydrology. But despite these developments—mostly over the past decade—many technical problems still exist in terms of linking xylem water to its soil water and groundwater sources. This is mainly due to sampling issues and possible fractionation of xylem water. Here we explore whether or not leaf water alone can be used to quantify the blend of rainfall event inputs from which the leaf water originates. Leaf water has historically been avoided in plant water uptake studies due to the extreme fractionation processes at the leaf surface. In our proof of concept work we embrace those processes and use the well-known Craig and Gordon model to map leaf water back to its individual precipitation event water sources. We also employ a Bayesian uncertainty estimation approach to quantify source apportionment uncertainties. We show this using a controlled, vegetated lysimeter experiment where we were able to use leaf water to correctly identify the mean seasonal rainfall that was taken up by the plant, with an uncertainty typically within ±1‰ for δ18O. While not appropriate for all source water studies, this work shows that leaf water isotope composition may provide a new, relatively un-intrusive method for addressing questions about the plant water source.

Closure of the soil water balance is fundamental to ecohydrology. But closing the soil water balance with hydrometric information offers no insight into the age distribution of water transiting the soil column via deep drainage or the combination of soil evaporation and transpiration. This is a major challenge in our discipline currently; tracing the water balance is the needed next step. Here we report results from a controlled tracer experiment aimed at both closing the soil water balance and tracing its individual components. This was carried out on a 2.5 m3 lysimeter planted with a willow tree. We applied 25 mm of isotopically enriched water on top of the lysimeter and tracked it for 43 days through the soil water, the bottom drainage, and the plant xylem. We then destructively sampled the system to quantify the remaining isotope mass. More than 900 water samples were collected for stable isotope analysis to trace the labeled irrigation. We then used these data to quantify when and where the labeled irrigation became the source of plant uptake or deep percolation. Evapotranspiration dominated the water balance outflow (88%). Tracing the transpiration flux showed further that transpiration was soil water that had fallen as precipitation 1–2 months prior. The tracer breakthrough in transpiration was complex and different from the breakthrough curves observed within the soil or in the bottom drainage. Given the lack of direct experimental data on travel time to transpiration, these results provide a first balance closure where all the relevant outflows are traced.

Rationale Hydrogen and oxygen stable isotope ratios (δ2H, δ17O, and δ18O values) are commonly used tracers of water. These ratios can be measured by isotope ratio infrared spectroscopy (IRIS). However, IRIS approaches are prone to errors induced by organic compounds present in plant, soil, and natural water samples. A novel approach using 17O-excess values has shown promise for flagging spectrally contaminated plant samples during IRIS analysis. A systematic assessment of this flagging system is needed to prove it useful. Methods Errors induced by methanol and ethanol water mixtures on measured IRIS and isotope ratio mass spectrometry (IRMS) results were evaluated. For IRIS analyses both liquid- and vapour-mode (via direct vapour equilibration) methods are used. The δ2H, δ17O, and δ18O values were measured and compared with known reference values to determine the errors induced by methanol and ethanol contamination. In addition, the 17O-excess contamination detection approach was tested. This is a post-processing detection tool for both liquid and vapour IRIS triple-isotope analyses, utilizing calculated 17O-excess values to flag contaminated samples. Results Organic contamination induced significant errors in IRIS results, not seen in IRMS results. Methanol caused larger errors than ethanol. Results from vapour-IRIS analyses had larger errors than those from liquid-IRIS analyses. The 17O-excess approach identified methanol driven error in liquid- and vapour-mode IRIS samples at levels where isotope results became unacceptably erroneous. For ethanol contaminated samples, a mix of erroneous and correct flagging occurred with the 17O-excess method. Our results indicate that methanol is the more problematic contaminant for data corruption. The 17O-excess method was therefore useful for data quality control. Conclusions Organic contamination caused significant errors in IRIS stable isotope results. These errors were larger during vapour analyses than during liquid IRIS analyses, and larger for methanol than ethanol contamination. The 17O-excess method is highly sensitive for detecting narrowband (methanol) contamination error in vapour and liquid analysis modes in IRIS.

The boreal forest is a major contributor to the global climate system, therefore, reducing uncertainties in how the forest will respond to a changing climate is critical. One source of uncertainty is the timing and drivers of the spring transition. Remote sensing can provide important information on this transition, but persistent foliage greenness, seasonal snow cover, and a high prevalence of mixed forest stands (both deciduous and evergreen species) complicate interpretation of these signals. We collected tower-based remotely sensed data (reflectance-based vegetation indices and Solar-Induced Chlorophyll Fluorescence [SIF]), stem radius measurements, gross primary productivity, and environmental conditions in a boreal mixed forest stand. Evaluation of this data set shows a two-phased spring transition. The first phase is the reactivation of photosynthesis and transpiration in evergreens, marked by an increase in relative SIF, and is triggered by thawed stems, warm air temperatures, and increased available soil moisture. The second phase is a reduction in bulk photoprotective pigments in evergreens, marked by an increase in the Chlorophyll-Carotenoid Index. Deciduous leaf-out occurs during this phase, marked by an increase in all remotely sensed metrics. The second phase is controlled by soil thaw. Our results demonstrate that remote sensing metrics can be used to detect specific physiological changes in boreal tree species during the spring transition. The two-phased transition explains inconsistencies in remote sensing estimates of the timing and drivers of spring recovery. Our results imply that satellite-based observations will improve by using a combination of vegetation indices and SIF, along with species distribution information.

The stable isotopes of hydrogen and oxygen (δ2H and δ18O, respectively) have been widely used to investigate tree water source partitioning. These tracers have shed new light on patterns of tree water use in time and space. However, there are several limiting factors to this methodology (e.g., the difficult assessment of isotope fractionation in trees, and the labor‐intensity associated with the collection of significant sample sizes) and the use of isotopes alone has not been enough to provide a mechanistic understanding of source water partitioning. Here, we combine isotope data in xylem and soil water with measurements of tree's physiological information including tree water deficit (TWD), fine root distribution, and soil matric potential, to investigate the mechanism driving tree water source partitioning. We used a 2 m3 lysimeter with willow trees (Salix viminalis) planted within, to conduct a high spatial–temporal resolution experiment. TWD provided an integrated response of plant water status to water supply and demand. The combined isotopic and TWD measurement showed that short‐term variation (within days) in source water partitioning is determined mainly by plant hydraulic response to changes in soil matric potential. We observed changes in the relationship between soil matric potential and TWD that are matched by shifts in source water partitioning. Our results show that tree water use is a dynamic process on the time scale of days. These findings demonstrate tree's plasticity to water supply over days can be identified with high‐resolution measurements of plant water status. Our results further support that root distribution alone is not an indicator of water uptake dynamics. Overall, we show that combining physiological measurements with traditional isotope tracing can reveal mechanistic insights into plant responses to changing environmental conditions.