Growth ranking of hybrid aspen genotypes and its linkage to leaf gas exchange.

BACKGROUND: Afforestation of non-forestland is a new measure by the European Union to enhance climate mitigation and biodiversity. Hybrid aspen (Populus tremula L. × P. tremuloides Michx.) is among the suitable tree species for afforestation to produce woody biomass. However, the best performing genotypic material for intensive biomass production and its physiological adaptation capacity is still unclear. We compared 22 hybrid aspen genotypes growth and leaf physiological characteristics (stomatal conductance, net photosynthesis, intrinsic water-use efficiency) according to their geographical north- or southward transfer (European P. tremula parent from 51° to 60° N and North American P. tremuloides parent from 45° to 54° N) to hemiboreal Estonia (58° N) in a completely randomized design progeny trial. We tested whether the growth ranking of genotypes of different geographical origin has changed from young (3-year-old) to mid-rotation age (13-year-old). The gas exchange parameters were measured in excised shoots in 2021 summer, which was characterised with warmer (+ 4 °C) and drier (17% precipitation from normal) June and July than the long-term average. RESULTS: We found that the northward transfer of hybrid aspen genotypes resulted in a significant gain in growth (two-fold greater diameter at breast height) in comparison with the southward transfer. The early selection of genotypes was generally in good accordance with the middle-aged genotype ranking, while some of the northward transferred genotypes showed improved growth at the middle-age period in comparison with their ranking during the early phase. The genotypes of southward transfer demonstrated higher stomatal conductance, which resulted in higher net photosynthesis, and lower intrinsic water-use efficiency (iWUE) compared with northward transfer genotypes. However, higher photosynthesis did not translate into higher growth rate. The higher physiological activity of southern transferred genotypes was likely related to a better water supply of smaller and consequently more shaded trees under drought. Leaf nitrogen concentration did not have any significant relation with tree growth. CONCLUSIONS: We conclude that the final selection of hybrid aspen genotypes for commercial use should be done in 10-15 years after planting. Physiological traits acquired during periods of droughty conditions may not fully capture the growth potential. Nonetheless, we advocate for a broader integration of physiological measurements alongside traditional traits (such as height and diameter) in genotype field testing to facilitate the selection of climate-adapted planting material for resilient forests.

Effects of air humidity and soil moisture on secondary metabolites in the leaves and roots of Betula pendula of different competitive status.

Plant secondary metabolites (PSMs) defend plants against abiotic stresses, including those caused by climate change and against biotic stresses, such as herbivory and competition. There is a trade-off between allocating available carbon to growth and defence in stressful environments. However, our knowledge about trade-off is limited, especially when abiotic and biotic stresses co-occur. We aimed to understand the combined effect of increasing precipitation and humidity, the tree's competitive status, and canopy position on leaf secondary metabolites (LSMs) and fine root secondary metabolites (RSMs) in Betula pendula. We sampled 8-year-old B. pendula trees growing in the free air humidity manipulation (FAHM) experimental site, where treatments included elevated relative air humidity and elevated soil moisture. A high-performance liquid chromatography-quadrupole-time of flight mass spectrometer (HPLC-qTOF-MS) was used to analyse secondary metabolites. Our results showed accumulation of LSM depends on the canopy position and competitive status. Flavonoids (FLA), dihydroxybenzoic acids (HBA), jasmonates (JA) and terpene glucosides (TG) were higher in the upper canopy, and FLA, monoaryl compounds (MAR) and sesquiterpenoids (ST) were higher in dominant trees. The FAHM treatments had a more distinct effect on RSM than on LSM. The RSMs were lower in elevated air humidity and soil moisture conditions than in control conditions. The RSM content depended on the competitive status and was higher in suppressed trees. Our study suggests that young B. pendula will allocate similar amounts of carbon to constitutive chemical leaf defence, but a lower amount to root defence (per fine root biomass) under higher humidity.

Night-time water relations and gas exchange in cut shoots of five boreal dwarf shrub species: impact of soil water availability.

Recent findings suggest that drought may affect plants' daytime and night-time stomatal regulation differently. However, knowledge of night-time stomatal behaviour in dwarf shrubs growing in boreal ecosystems is lacking. We sampled cut shoots from dwarf shrub species to elucidate their capacity to transpire at night and the effect of drought on stomatal regulation. The shoots' water relations and gas exchange were measured under controlled conditions in a growth chamber. The studied species demonstrated considerable differences in their diurnal water use. The night-time water use percentage of daytime water use (NWU) reached up to 90% in Andromeda polifolia and Vaccinium uliginosum. In Rhododendron tomentosum, Vaccinium myrtillus and Chamaedaphne calyculata, the NWU was 62, 27 and 26%, respectively. The shoots of C. calyculata showed a significant increase (P < 0.001) in the transpiration rate (E) during the night. However, in R. tomentosum, a decrease (P < 0.05) in nightly E was observed. The shoot conductance (g) at the end of the night was lower than daytime g in all studied species, but the difference was not significant for V. uliginosum. Across the species, NWU was negatively related (P < 0.001) to the soil volumetric water content (SWC) in the plant habitat. However, daytime E and g were positively related (P < 0.05) to the habitat SWC. Only in V. myrtillus was night-time E higher (P < 0.05) in dry conditions than in wet conditions. Our results demonstrate high variability in diurnal water relations in dwarf shrubs, which can keep stomata open in the dark even when drought limits daytime g and E.

The competitive status of trees determines their responsiveness to increasing atmospheric humidity - a climate trend predicted for northern latitudes.

The interactive effects of climate variables and tree-tree competition are still insufficiently understood drivers of forest response to global climate change. Precipitation and air humidity are predicted to rise concurrently at high latitudes of the Northern Hemisphere. We investigated whether the growth response of deciduous trees to elevated air humidity varies with their competitive status. The study was conducted in seed-originated silver birch and monoclonal hybrid aspen stands grown at the free air humidity manipulation (FAHM) experimental site in Estonia, in which manipulated stands (n = 3 for both species) are exposed to artificially elevated relative air humidity (6-7% over the ambient level). The study period included three growing seasons during which the stands had reached the competitive stage (trees were 7 years old in the final year). A significant 'treatment×competitive status' interactive effect on growth was detected in all years in birch (P < 0.01) and in one year in aspen stands (P = 0.015). Competitively advantaged trees were always more strongly affected by elevated humidity. Initially the growth of advantaged and neutral trees of both species remained significantly suppressed in humidified stands. In the following years, dominance and elevated humidity had a synergistic positive effect on the growth of birches. Aspens with different competitive status recovered more uniformly, attaining similar relative growth rates in manipulated and control stands, but preserved a significantly lower total growth yield due to severe initial growth stress. Disadvantaged trees of both species were never significantly affected by elevated humidity. Our results suggest that air humidity affects trees indirectly depending on their social status. Therefore, the response of northern temperate and boreal forests to a more humid climate in future will likely be modified by competitive relationships among trees, which may potentially affect species composition and cause a need to change forestry practices.

Rapid and long-term effects of water deficit on gas exchange and hydraulic conductance of silver birch trees grown under varying atmospheric humidity.

BACKGROUND: Effects of water deficit on plant water status, gas exchange and hydraulic conductance were investigated in Betula pendula under artificially manipulated air humidity in Eastern Estonia. The study was aimed to broaden an understanding of the ability of trees to acclimate with the increasing atmospheric humidity predicted for northern Europe. Rapidly-induced water deficit was imposed by dehydrating cut branches in open-air conditions; long-term water deficit was generated by seasonal drought. RESULTS: The rapid water deficit quantified by leaf (ΨL) and branch water potentials (ΨB) had a significant (P < 0.001) effect on gas exchange parameters, while inclusion of ΨB in models resulted in a considerably better fit than those including ΨL, which supports the idea that stomatal openness is regulated to prevent stem rather than leaf xylem dysfunction. Under moderate water deficit (ΨL≥-1.55 MPa), leaf conductance to water vapour (gL), transpiration rate and leaf hydraulic conductance (KL) were higher (P < 0.05) and leaf temperature lower in trees grown in elevated air humidity (H treatment) than in control trees (C treatment). Under severe water deficit (ΨL<-1.55 MPa), the treatments showed no difference. The humidification manipulation influenced most of the studied characteristics, while the effect was to a great extent realized through changes in soil water availability, i.e. due to higher soil water potential in H treatment. Two functional characteristics (gL, KL) exhibited higher (P < 0.05) sensitivity to water deficit in trees grown under increased air humidity. CONCLUSIONS: The experiment supported the hypothesis that physiological traits in trees acclimated to higher air humidity exhibit higher sensitivity to rapid water deficit with respect to two characteristics - leaf conductance to water vapour and leaf hydraulic conductance. Disproportionate changes in sensitivity of stomatal versus leaf hydraulic conductance to water deficit will impose greater risk of desiccation-induced hydraulic dysfunction on the plants, grown under high atmospheric humidity, in case of sudden weather fluctuations, and might represent a potential threat in hemiboreal forest ecosystems. There is no trade-off between plant hydraulic capacity and photosynthetic water-use efficiency on short time scale.

Dry and wet periods determine stem and soil greenhouse gas fluxes in a northern drained peatland forest.

Greenhouse gas (GHG) fluxes from peatland soils are relatively well studied, whereas tree stem fluxes have received far less attention. Simultaneous year-long measurements of soil and tree stem GHG fluxes in northern peatland forests are scarce, as previous studies have primarily focused on the growing season. We determined the seasonal dynamics of tree stem and soil CH4, N2O and CO2 fluxes in a hemiboreal drained peatland forest. Gas samples for flux calculations were manually collected from chambers at different heights on Downy Birch (Betula pubescens) and Norway Spruce (Picea abies) trees (November 2020-December 2021) and analysed using gas chromatography. Environmental parameters were measured simultaneously with fluxes and xylem sap flow was recorded during the growing season. Birch stems played a greater role in the annual GHG dynamics than spruce stems. Birch stems were net annual CH4, N2O and CO2 sources, while spruce stems constituted a CH4 and CO2 source but a N2O sink. Soil was a net CO2 and N2O source, but a sink of CH4. Temporal dynamics of stem CH4 and N2O fluxes were driven by isolated emissions' peaks that contributed significantly to net annual fluxes. Stem CO2 efflux followed a seasonal trend coinciding with tree growth phenology. Stem CH4 dynamics were significantly affected by the changes between wetter and drier periods, while N2O was more influenced by short-term changes in soil hydrologic conditions. We showed that CH4 emitted from tree stems during the wetter period can offset nearly half of the soil sink capacity. We presented for the first time the relationship between tree stem GHG fluxes and sap flow in a peatland forest. The net CH4 flux was likely an aggregate of soil-derived and stem-produced CH4. A dominating soil source was more evident for stem N2O fluxes.

Responses of fine root exudation, respiration and morphology in three early successional tree species to increased air humidity and different soil nitrogen sources.

Global climate change scenarios predict an increase in air temperature, precipitation and air humidity for northern latitudes. Elevated air humidity may significantly reduce the water flux through forest canopies and affect interactions between water and nutrient uptake. However, we have limited understanding of how altered transpiration would affect root respiration and carbon (C) exudation as fine root morphology acclimates to different water flux. We investigated the effects of elevated air relative humidity (eRH) and different inorganic nitrogen sources (NO3- and NH4+) on above and belowground traits in hybrid aspen (Populus × wettsteinii Hämet-Ahti), silver birch (Betula pendula Roth.) and Scots pine (Pinus sylvestris L.) grown under controlled climate chamber conditions. The eRH significantly decreased the transpiration flux in all species, decreased root mass-specific exudation in pine, and increased root respiration in aspen. eRH also affected fine root morphology, with specific root area increasing for birch but decreasing in pine. The species comparison revealed that pine had the highest C exudation, whereas birch had the highest root respiration rate. Both humidity and nitrogen treatments affected the share of absorptive and pioneer roots within fine roots; however, the response was species-specific. The proportion of absorptive roots was highest in birch and aspen, the share of pioneer roots was greatest in aspen and the share of transport roots was greatest in pine. Fine roots with lower root tissue density were associated with pioneer root tips and had a higher C exudation rate. Our findings underline the importance of considering species-specific differences in relation to air humidity and soil nitrogen availability that interactively affect the C input-output balance. We highlight the role of changes in the fine root functional distribution as an important acclimation mechanism of trees in response to environmental change.

Predawn leaf conductance depends on previous day irradiance but is not related to growth in aspen saplings grown under artificially manipulated air humidity.

Recent studies have suggested that predawn stomatal opening may enhance early-morning photosynthesis (A) and improve the relative growth rate of trees. However, the causality between night-time stomatal conductance, A, and tree growth is disputable because stomatal opening in darkness can be mediated by previous day photosynthate loads and might be a consequence of growth-related processes like dark respiration (R). To identify linkages between night-time leaf conductance (gl_night), A, R, and tree growth, we conducted an experiment in hybrid aspen saplings grown under different air relative humidity (RH) conditions and previous day irradiance level (IR_pday). Predawn leaf conductance (gl_predawn) depended on RH, IR_pday and R (P < 0.05), whereas early-morning gross A (Agross_PAR500) depended on IR_pday and gl_predawn (P < 0.001). Daytime net A was positively related to Agross_PAR500 and leaf [N] (P < 0.05). Tree diameter and height increment correlated positively with gl at the beginning and middle of the night (P < 0.05) but not before dawn. Although our results demonstrate that gl_night was related to tree growth, the relationship was not determined by R. The linkage between gl_predawn and Agross_PAR500 was modified by IR_pday, indicating that daily CO2 assimilation probably provides feedback for stomatal opening before dawn.

Effects of light intensity and duration on leaf hydraulic conductance and distribution of resistance in shoots of silver birch (Betula pendula).

Variation in leaf hydraulic conductance (K(L)) and distribution of resistance in response to light intensity and duration were examined in shoots of silver birch (Betula pendula Roth). K(L) was determined on detached shoots using the evaporative flux method (transpiration was measured with a porometer and water potential drop with a pressure chamber). Although K(L) depended on light duration per se, its dynamics was largely determined by leaf temperature (T(L)). Both upper-crown leaves and branches developed in well-illuminated environment exhibited higher hydraulic efficiency compared with the lower crown, accounting for vertical trends of apparent soil-to-leaf hydraulic conductance in canopy of silver birch revealed in our previous studies. K(L) varied significantly with light intensity, the highest values for both shade and sun foliage were recorded at photosynthetic photon flux density of 330 micromol m(-2) s(-1). Light responses of K(L) were associated evidently with an irradiance-mediated effect on extravascular tissues involving regulation of cell membrane aquaporins. Effects of irradiance on K(L) resulted in changes of Psi(L), bringing about considerable alteration in partitioning of the resistance between leaves and branch (leafless shoot stem): the contribution of leaves to the shoot total resistance decreased from 94% at -1.0 MPa to 75% at -0.2 MPa. Treatment with HgCl2 decreased hydraulic conductance of both leaves and branches, implying that condition of bordered pit membranes or shoot living tissues may be involved in responses of xylem conductance to Hg2+.

Temperature, light and leaf hydraulic conductance of little-leaf linden (Tilia cordata) in a mixed forest canopy.

Response of whole-leaf hydraulic conductance (G(L)) in little-leaf linden (Tilia cordata Mill.) to temperature and photosynthetic photon flux (Q(P)) was estimated by the evaporative flux method under natural conditions in a mixed forest canopy. Mean midday G(L) in the lower- and upper-crown foliage was 1.14 and 3.06 mmol m(-2) s(-1) MPa(-1), respectively. Over the study period, leaf temperature (T(L)) explained about 67% of the variation in G(L), and Q(P) explained about 10%. Leaf water potential and crown position also affected G(L) significantly. About a third of the temperature effect was attributable to changes in the viscosity of water, and two thirds to changes in protoplast permeability (i.e., symplastic conductance). Leaf hydraulic conductance was highly sensitive to changes in Q(P) when Q(P) was less than 200 micromol m(-2) s(-1), and G(L) sensitivity decreased with increasing irradiance. Sensitivity of G(L) to variation in T(L) increased consistently with increasing temperature in the range of 16 to 29 degrees C. There were positive interactions between temperature and light in their effects on G(L): the light response was more pronounced at higher leaf temperatures. Because of frequent rains during the study period, the trees experienced no soil water deficit, and, within the range experienced, soil water potential had no effect on G(L). Leaf hydraulic conductance exhibited a seasonal pattern that could be explained primarily by temporal variability in mean air temperature and irradiance, in addition to which an age-related trend (P<0.001) of increasing G(L) from the end of June to the beginning of August was observed.

Increasing air humidity influences hydraulic efficiency but not functional vulnerability of xylem in hybrid aspen.

Climate models predict greater increases in the frequency than in the amount of precipitation and a consequent rise in atmospheric humidity at high latitudes by the end of the century. We investigated the responses of hydraulic and relevant anatomical traits of xylem to elevated relative humidity of air on a 1-yr-old coppice of hybrid aspen (Populus×wettsteinii) growing in the experimental stand at the Free Air Humidity Manipulation site in Eastern Estonia. The hydraulic conductivity of stems was measured with a high pressure flow meter; artificial cavitation in the stem segments was induced by the air injection method. Specific conductivity of xylem decreased from 4.42 in the control to 3.94kgm-1s-1MPa-1 in the humidification treatment, while the trend was well correlated with increasing wood density. Humidified trees exhibited smaller leaf area at the same xylem cross-sectional area, resulting in 34% higher average Huber values compared to the control. Control and humidity-treated trees differed by neither native embolism level nor susceptibility to dehydration-induced cavitation. Increasing atmospheric humidity reduces the hydraulic efficiency of hybrid aspen trees expressed on a xylem area basis and causes substantial changes in resource allocation between photosynthetic and water transport tissues. This climate trend does not influence stem vulnerability to cavitation.

Elevated Air Humidity Changes Soil Bacterial Community Structure in the Silver Birch Stand.

Soil microbes play a fundamental role in forest ecosystems and respond rapidly to changes in the environment. Simultaneously with the temperature increase the climate change scenarios also predict an intensified hydrological cycle for the Baltic Sea runoff region. The aim of this study was to assess the effect of elevated air humidity on the top soil microbial community structure of a silver birch (Betula pendula Roth.) stand by using a free air humidity manipulation facility (FAHM). The bacterial community structures of bulk soil and birch rhizosphere were analyzed using high-throughput sequencing of bacteria-specific16S rRNA gene fragments and quantification of denitrification related genes. The increased air humidity altered both bulk soil and rhizosphere bacterial community structures, and changes in the bacterial communities initiated by elevated air humidity were related to modified soil abiotic and biotic variables. Network analysis revealed that variation in soil bacterial community structural units is explained by altered abiotic conditions such as increased pH value in bulk soil, while in rhizosphere the change in absorptive root morphology had a higher effect. Among root morphological traits, the absorptive root diameter was strongest related to the bacterial community structure. The changes in bacterial community structures under elevated air humidity are associated with shifts in C, N, and P turnover as well as mineral weathering processes in soil. Increased air humidity decreased the nir and nosZ gene abundance in the rhizosphere bacterial community. The potential contribution of the denitrification to the N2O emission was not affected by the elevated air humidity in birch stand soil. In addition, the study revealed a strong link between the bacterial community structure, abundance of denitrification related genes, and birch absorptive root morphology in the ecosystem system adaptation to elevated air humidity.

Responses of sap flow, leaf gas exchange and growth of hybrid aspen to elevated atmospheric humidity under field conditions.

An increase in average air temperature and frequency of rain events is predicted for higher latitudes by the end of the 21st century, accompanied by a probable rise in air humidity. We currently lack knowledge on how forest trees acclimate to rising air humidity in temperate climates. We analysed the leaf gas exchange, sap flow and growth characteristics of hybrid aspen (Populus tremula × P. tremuloides) trees growing at ambient and artificially elevated air humidity in an experimental forest plantation situated in the hemiboreal vegetation zone. Humidification manipulation did not affect the photosynthetic capacity of plants, but did affect stomatal responses: trees growing at elevated air humidity had higher stomatal conductance at saturating photosynthetically active radiation (gs sat) and lower intrinsic water-use efficiency (IWUE). Reduced stomatal limitation of photosynthesis in trees grown at elevated air humidity allowed slightly higher net photosynthesis and relative current-year height increments than in trees at ambient air humidity. Tree responses suggest a mitigating effect of higher air humidity on trees under mild water stress. At the same time, trees at higher air humidity demonstrated a reduced sensitivity of IWUE to factors inducing stomatal closure and a steeper decline in canopy conductance in response to water deficit, implying higher dehydration risk. Despite the mitigating impact of increased air humidity under moderate drought, a future rise in atmospheric humidity at high latitudes may be disadvantageous for trees during weather extremes and represents a potential threat in hemiboreal forest ecosystems.

Climate change at northern latitudes: rising atmospheric humidity decreases transpiration, N-uptake and growth rate of hybrid aspen.

At northern latitudes a rise in atmospheric humidity and precipitation is predicted as a consequence of global climate change. We studied several growth and functional traits of hybrid aspen (Populus tremula L.×P. tremuloides Michx.) in response to elevated atmospheric humidity (on average 7% over the ambient level) in a free air experimental facility during three growing seasons (2008-2010) in Estonia, which represents northern temperate climate (boreo-nemoral zone). Data were collected from three humidified (H) and three control (C) plots, and analysed using nested linear models. Elevated air humidity significantly reduced height, stem diameter and stem volume increments and transpiration of the trees whereas these effects remained highly significant also after considering the side effects from soil-related confounders within the 2.7 ha study area. Tree leaves were smaller, lighter and had lower leaf mass per area (LMA) in H plots. The magnitude and significance of the humidity treatment effect--inhibition of above-ground growth rate--was more pronounced in larger trees. The lower growth rate in the humidified plots can be partly explained by a decrease in transpiration-driven mass flow of NO(3) (-) in soil, resulting in a significant reduction in the measured uptake of N to foliage in the H plots. The results suggest that the potential growth improvement of fast-growing trees like aspens, due to increasing temperature and atmospheric CO(2) concentration, might be smaller than expected at high latitudes if a rise in atmospheric humidity simultaneously takes place.

Do stomata operate at the same relative opening range along a canopy profile of Betula pendula?

Stomatal density and size were measured along the light gradient of a Betula pendula Roth. canopy in relation to microclimatic conditions. The theoretical stomatal conductance was calculated using stomatal density and dimensions to predict to what degree stomatal conductance is related to anatomical properties and relative stomatal opening. Stomatal density was higher and leaf area smaller in the upper canopy, whereas epidermal cell density did not change significantly along the canopy light gradient, indicating that stomatal initiation is responsible for differences in stomatal density. Stomatal dimensions - the length of guard cell on the dorsal side and the guard cell width - decreased with declining light availability. Maximum measured stomatal conductance and modelled stomatal conductance were higher at the top of the crown. The stomata operate closer to their maximum openness and stomatal morphology is a more important determinant of stomatal conductance in the top leaves than in leaves of lower canopy. As stomata usually limit photosynthesis more in upper than in lower canopy, it was concluded that stomatal morphology can principally be important for photosynthesis limitation in upper canopy.

Effects of light availability versus hydraulic constraints on stomatal responses within a crown of silver birch.

Responses of leaf conductance (gL) to variation in photosynthetic photon flux density (QP), leaf-to-air vapour pressure difference (VPD), bulk leaf water potential (Psi(x)), and total hydraulic conductance (GT) were examined in silver birch (Betula pendula Roth) with respect to leaf position in the crown. To reduce limitations caused by insufficient water supply or low light availability, experiments were also performed with branchlets cut from two different canopy layers. The intact upper-canopy leaves demonstrated 1.8-2.0 times higher (P<0.001) daily maxima of gL compared with the lower-canopy leaves growing in the shadow of upper branches. In the morning, gL in the shade foliage was primarily constrained by low light availability, in the afternoon, by limited water supply. Leaf conductance decreased when Psi(x) fell below certain values around midday, while the sun foliage experienced greater negative water potentials than the shade foliage. Midday stomatal openness was controlled by leaf water status and temperature, rather than by transpiration rate (E) via the feedforward mechanism. Mean GT was 1.7 times higher (P<0.001) for the upper-canopy foliage compared to that of the lower canopy. At least 34-39% of the total resistance to the water flow from soil up to the shade foliage, and 54% up to the sun foliage, resided in 30-cm distal parts of the branches. Artificial reduction of hydraulic constraints raised Psi(x) and made gL less sensitive to changes in both atmospheric and plant factors. Improved water supply increased gL and E in the lower-canopy foliage, but not in the upper-canopy foliage. The results support the idea that leaves in the lower canopy are hydraulically more constrained than in the upper canopy.