Estimating intraseasonal intrinsic water-use efficiency from high-resolution tree-ring delta C-13 data in boreal Scots pine forests

Intrinsic water-use efficiency (iWUE), a key index for carbon and water balance, has been widely estimated from tree-ring delta C-13 at annual resolution, but rarely at high-resolution intraseasonal scale. We estimated high-resolution iWUE from laser-ablation delta C-13 analysis of tree-rings (iWUE(iso)) and compared it with iWUE derived from gas exchange (iWUE(gas)) and eddy covariance (iWUE(EC)) data for two Pinus sylvestris forests from 2002 to 2019. By carefully timing iWUE(iso) via modeled tree-ring growth, iWUE(iso) aligned well with iWUE(gas) and iWUE(EC) at intraseasonal scale. However, year-to-year patterns of iWUE(gas), iWUE(iso), and iWUE(EC) were different, possibly due to distinct environmental drivers on iWUE across leaf, tree, and ecosystem scales. We quantified the modification of iWUE(iso) by postphotosynthetic delta C-13 enrichment from leaf sucrose to tree rings and by nonexplicit inclusion of mesophyll and photorespiration terms in photosynthetic discrimination model, which resulted in overestimation of iWUE(iso) by up to 11% and 14%, respectively. We thus extended the application of tree-ring delta C-13 for iWUE estimates to high-resolution intraseasonal scale. The comparison of iWUE(gas), iWUE(iso), and iWUE(EC) provides important insights into physiological acclimation of trees across leaf, tree, and ecosystem scales under climate change and improves the upscaling of ecological models.Peer reviewe

Estimating intra-seasonal photosynthetic discrimination and water use efficiency using delta C-13 of leaf sucrose in Scots pine

Comparison of assimilate delta C-13 values estimated from different leaf carbon pools, Picarro measurements, and modelled data demonstrates the potential for misinterpretation arising from bulk organic matter delta C-13 analysis.Sucrose has a unique role in recording environmental and physiological signals during photosynthesis in its carbon isotope composition (delta C-13) and transport of the signal to tree rings. Yet, instead of sucrose, total organic matter (TOM) or water-soluble carbohydrates (WSC) are typically analysed in studies that follow delta C-13 signals within trees. To study how the choice of organic material may bias the interpretation of delta C-13 records, we used mature field-grown Scots pine (Pinus sylvestris) to compare for the first time delta C-13 of different leaf carbon pools with delta C-13 of assimilates estimated by a chamber-Picarro system (delta C-13(A_Picarro)), and a photosynthetic discrimination model (delta C-13(A_model)). Compared with sucrose, the other tested carbon pools, such as TOM and WSC, poorly recorded the seasonal trends or absolute values of delta C-13(A_Picarro) and delta C-13(A_model). Consequently, in comparison with the other carbon pools, sucrose delta C-13 was superior for reconstructing changes in intrinsic water use efficiency (iWUE), agreeing in both absolute values and intra-seasonal variations with iWUE estimated from gas exchange. Thus, deriving iWUE and environmental signals from delta C-13 of bulk organic matter can lead to misinterpretation. Our findings underscore the advantage of using sucrose delta C-13 to understand plant physiological responses in depth.Peer reviewe

Explicitly accounting for needle sugar pool size crucial for predicting intra-seasonal dynamics of needle carbohydrates delta O-18 and delta C-13

We explore needle sugar isotopic compositions (delta O-18 and delta C-13) in boreal Scots pine (Pinus sylvestris) over two growing seasons. A leaf-level dynamic model driven by environmental conditions and based on current understanding of isotope fractionation processes was built to predict delta O-18 and delta C-13 of two hierarchical needle carbohydrate pools, accounting for the needle sugar pool size and the presence of an invariant pinitol pool. Model results agreed well with observed needle water delta O-18, delta O-18 and delta C-13 of needle water-soluble carbohydrates (sugars + pinitol), and needle sugar delta C-13 (R-2 = 0.95, 0.84, 0.60, 0.73, respectively). Relative humidity (RH) and intercellular to ambient CO2 concentration ratio (C-i/C-a) were the dominant drivers of delta O-18 and delta C-13 variability, respectively. However, the variability of needle sugar delta O-18 and delta C-13 was reduced on diel and intra-seasonal timescales, compared to predictions based on instantaneous RH and C-i/C-a, due to the large needle sugar pool, which caused the signal formation period to vary seasonally from 2 d to more than 5 d. Furthermore, accounting for a temperature-sensitive biochemical O-18-fractionation factor and mesophyll resistance in C-13-discrimination were critical. Interpreting leaf-level isotopic signals requires understanding on time integration caused by mixing in the needle sugar pool.Peer reviewe

Effects of variable temperature and moisture conditions on respiration and nonstructural carbohydrate dynamics of tree roots

In warming climates, soil water content (SWC) may act as an important factor in determining belowground carbon dynamics in boreal forests. Here, we estimated the respiration and nonstructural carbohydrate (NSC) concentrations of tree roots in a mature Scots pine (Pinus sylvestris L.) stand in southern Finland during two growing seasons with contrasting weather conditions. Root respiration was estimated with four different methods: 1) incubating excised roots, 2) partitioning forest floor respirations with root exclusion, or 3) based on temperature response functions and 4) modelling with the whole-tree carbon model 'CASSIA'. In addition, we conducted a drought experiment in a greenhouse to determine the effect of reduced soil-water availability on respiration by incubating soil and roots of Scots pine saplings. We observed that the respiration of incubated roots of Scots pine saplings and soil decreased with drying after excluding the effect of temperature on respiration (RRES), soil being more sensitive to drought than roots. Similarly, RRES of incubated roots in the field was significantly decreased by lowered SWC, whereas respiration of the entire root system estimated with other methods was clearly higher in dryer and warmer than moister and cooler year. Nevertheless, incubated roots excavated from the topsoil are most affected by drying soil, which might not reflect the response of the entire root system. RRES of incubated roots was negatively associated with root fructose and glucose concentrations. At the same time, root fructose, glucose and sucrose concentrations were negatively associated with SWC due to their role in osmoregulation. Thereby it seems that RRES does not directly follow the changes in NSCs despite the apparent correlation. Our study highlights the responsive nature of root carbon dynamics in varying weather events that should be taken into account in estimating and modelling the impacts of warming climate.Peer reviewe

Tree organ growth and carbon allocation dynamics impact the magnitude and δ13C signal of stem and soil CO2 fluxes

Incomplete knowledge of carbon (C) allocation dynamics in trees hinders accurate modeling and future predictions of tree growth. We studied C allocation dynamics in a mature Pinus sylvestris L. dominated forest with a novel analytical approach, allowing the first comparison of: (i) magnitude and delta C-13 of shoot, stem and soil CO2 fluxes (A(shoot), R-stem and R-soil), (ii) concentration and delta C-13 of compound-specific and/or bulk non-structural carbohydrates (NSCs) in phloem and roots and (iii) growth of stem and fine roots. Results showed a significant effect of phloem NSC concentrations on tracheid growth, and both variables significantly impacted R-stem. Also, concentrations of root NSCs, especially starch, had a significant effect on fine root growth, although no effect of root NSC concentrations or root growth was detected on R-soil. Time series analysis between delta C-13 of A(shoot) and delta C-13 of R-stem or delta C-13 of R-soil revealed strengthened C allocation to stem or roots under high C demands. Furthermore, we detected a significant correlation between delta C-13 of R-stem and delta C-13 of phloem sucrose and glucose, but not for starch or water-soluble carbohydrates. Our results indicate the need to include C allocation dynamics into tree growth models. We recommend using compound-specific concentration and delta C-13 analysis to reveal C allocation processes that may not be detected by the conventional approach that utilizes bulk organic matter.Peer reviewe

Fast recovery of suppressed Norway spruce trees after selection harvesting on a drained peatland forest site

Continuous cover forestry (CCF) has been promoted as an environmentally sustainable option for drained peatlands. The CCF management has been challenged due to potentially lower tree growth compared to tradi-tional even-aged management, especially with suppressed trees that are released during a selection harvesting under CCF management. Our objective was to quantify the time lag of stem diameter growth response of suppressed Norway spruce trees (Picea abies Karst.) after a selection harvesting compared to that of dominant trees. We also tested if the carbon assimilation of the trees increased immediately after selection harvesting. We used radial increment cores from suppressed Norway spruce trees to estimate the impact of selection harvesting on the diameter growth and intrinsic water use efficiency (iWUE). We measured carbon isotope composition (delta 13C) of wood, to quantify how the reduced competition between trees altered iWUE and its components, the photosynthetic rate (A) and sto-matal conductance (g). The study was conducted in the Lettosuo experimental site on fertile forestry drained peatland area in southern Finland. Approximately 70 % of the initial stand area (18.5 ha) was harvested according to CCF principles by applying selection harvesting, and the rest of the area was divided to intact control area and to clear-cut area. In the study site, by selection harvest, trees were removed from multiple age classes, but especially mature trees individually or in a small groups were taken away to maintain uneven-aged structure of the forest. All the target trees grew in the similar competitive position before selection harvesting. Our results show that there was a delay with the diameter growth of the suppressed trees to selection har-vesting, whereas the most significant growth-enhancing effect occurred three-four years after selection har-vesting. In contrast to the delay in the increment, the photosynthetic rate relative to stomatal conductance increased immediately after selection harvesting, as shown by the instant 2.5 parts per thousand increase in delta 13C to a post-harvest level. Our results show that carbon uptake increased immediately for suppressed Norway spruce trees after selection harvesting, but the harvest did not induce a clear increase in stem diameter growth during the first years after the harvest.Peer reviewe

Resin acid δ13C and δ18O as indicators of intra‐seasonal physiological and environmental variability

Understanding the dynamics of δ13C and δ18O in modern resin is crucial for interpreting (sub)fossilized resin records and resin production dynamics. We measured the δ13C and δ18O offsets between resin acids and their precursor molecules in the top-canopy twigs and breast-height stems of mature Pinus sylvestris trees. We also investigated the physiological and environmental signals imprinted in resin δ13C and δ18O at an intra-seasonal scale. Resin δ13C was c. 2‰ lower than sucrose δ13C, in both twigs and stems, likely due to the loss of 13C-enriched C-1 atoms of pyruvate during isoprene formation and kinetic isotope effects during diterpene synthesis. Resin δ18O was c. 20‰ higher than xylem water δ18O and c. 20‰ lower than δ18O of water-soluble carbohydrates, possibly caused by discrimination against 18O during O2-based diterpene oxidation and 35%–50% oxygen atom exchange with water. Resin δ13C and δ18O recorded a strong signal of soil water potential; however, their overall capacity to infer intraseasonal environmental changes was limited by their temporal, within-tree and among-tree variations. Future studies should validate the potential isotope fractionation mechanisms associated with resin synthesis and explore the use of resin δ13C and δ18O as a long-term proxy for physiological and environmental changes

The unknown third – Hydrogen isotopes in tree-ring cellulose across Europe

This is the first Europe-wide comprehensive assessment of the climatological and physiological information recorded by hydrogen isotope ratios in tree-ring cellulose (δ2Hc) based on a unique collection of annually resolved 100-year tree-ring records of two genera (Pinus and Quercus) from 17 sites (36°N to 68°N). We observed that the high-frequency climate signals in the δ2Hc chronologies were weaker than those recorded in carbon (δ13Cc) and oxygen isotope signals (δ18Oc) but similar to the tree-ring width ones (TRW). The δ2Hc climate signal strength varied across the continent and was stronger and more consistent for Pinus than for Quercus. For both genera, years with extremely dry summer conditions caused a significant 2H-enrichment in tree-ring cellulose. The δ2Hc inter-annual variability was strongly site-specific, as a result of the imprinting of climate and hydrology, but also physiological mechanisms and tree growth. To differentiate between environmental and physiological signals in δ2Hc, we investigated its relationships with δ18Oc and TRW. We found significant negative relationships between δ2Hc and TRW (7 sites), and positive ones between δ2Hc and δ18Oc (10 sites). The strength of these relationships was nonlinearly related to temperature and precipitation. Mechanistic δ2Hc models performed well for both genera at continental scale simulating average values, but they failed on capturing year-to-year δ2Hc variations. Our results suggest that the information recorded by δ2Hc is significantly different from that of δ18Oc, and has a stronger physiological component independent from climate, possibly related to the use of carbohydrate reserves for growth. Advancements in the understanding of 2H-fractionations and their relationships with climate, physiology, and species-specific traits are needed to improve the modelling and interpretation accuracy of δ2Hc. Such advancements could lead to new insights into trees' carbon allocation mechanisms, and responses to abiotic and biotic stress conditions

Newer Developments in Tree-Ring Stable Isotope Methods

The tree-ring stable C, O and H isotope compositions have proven valuable for examining past changes in the environment and predicting forest responses to environmental change. However, we have not yet recovered the full potential of this archive, partly due to a lack understanding of fractionation processes resulting from methodological constraints. With better understanding of the biochemical and tree physiological processes that lead to differences between the isotopic compositions of primary photosynthates and the isotopic compositions of substrates deposited in stem xylem, more reliable and accurate reconstructions could be obtained. Furthermore, by extending isotopic analysis of tree-ring cellulose to intra-molecular level, more information could be obtained on changing climate, tree metabolism or ecophysiology. This chapter presents newer methods in isotope research that have become available or show high future potential for fully utilising the wealth of information available in tree-rings. These include compound-specific analysis of sugars and cyclitols, high spatial resolution analysis of tree rings with UV-laser, and position-specific isotope analysis of cellulose. The aim is to provide the reader with understanding of the advantages and of the current challenges connected with the use of these methods for stable isotope tree-ring research