Making the Genotypic Variation Visible: Hyperspectral Phenotyping in Scots Pine Seedlings.

Hyperspectral reflectance contains valuable information about leaf functional traits, which can indicate a plant's physiological status. Therefore, using hyperspectral reflectance for high-throughput phenotyping of foliar traits could be a powerful tool for tree breeders and nursery practitioners to distinguish and select seedlings with desired adaptation potential to local environments. We evaluated the use of 2 nondestructive methods (i.e., leaf and proximal/canopy) measuring hyperspectral reflectance in the 350- to 2,500-nm range for phenotyping on 1,788 individual Scots pine seedlings belonging to lowland and upland ecotypes of 3 different local populations from the Czech Republic. Leaf-level measurements were collected using a spectroradiometer and a contact probe with an internal light source to measure the biconical reflectance factor of a sample of needles placed on a black background in the contact probe field of view. The proximal canopy measurements were collected under natural solar light, using the same spectroradiometer with fiber optical cable to collect data on individual seedlings' hemispherical conical reflectance factor. The latter method was highly susceptible to changes in incoming radiation. Both spectral datasets showed statistically significant differences among Scots pine populations in the whole spectral range. Moreover, using random forest and support vector machine learning algorithms, the proximal data obtained from the top of the seedlings offered up to 83% accuracy in predicting 3 different Scots pine populations. We conclude that both approaches are viable for hyperspectral phenotyping to disentangle the phenotypic and the underlying genetic variation within Scots pine seedlings.

Derivative-based time-adjusted analysis of diurnal and within-tree variation in the OJIP fluorescence transient of silver birch.

The JIP test, based on fast chlorophyll fluorescence (ChlF) kinetics and derived parameters, is a dependable tool for studying photosynthetic efficiency under varying environmental conditions. We extracted additional information from the whole OJIP and the normalized variable fluorescence (Vt) transient curve using first and second-order derivatives to visualize and localize points of landmark events. To account for light-induced variations in the fluorescence transient, we present a time-adjusted JIP test approach in which the derivatives of the transient curve are used to determine the exact timing of the J and I steps instead of fixed time points. We compared the traditional JIP test method with the time-adjusted method in analyzing fast ChlF measurements of silver birch (Betula pendula) in field conditions studying diurnal and within-crown variation. The time-adjusted JIP test method showed potential for studying ChlF dynamics, as it takes into account potential time shifts in the occurrence of J and I steps. The exact occurrence times of J and I steps and other landmark events coincided with the times of significant differences in fluorescence intensity. Chlorophyll fluorescence parameters were linearly related to photosynthetic photon flux density (PPFD) at different times of day, and the values obtained by the time-adjusted JIP test showed a stronger linear regression than the traditional JIP test. For fluorescence parameters having significant differences among different times of day and crown layers, the time-adjusted JIP test resulted in more clear differences than the traditional JIP test. Diurnal ChlF intensity data indicated that differences between the southern and northern provenance were only evident under low light conditions. Taken together, our results emphasize the potential relevance of considering the time domain in the analysis of the fast ChlF induction.

Polar day syndrome: differences in growth, photosynthetic traits and sink-size patterns between northern and southern Finnish silver birch (Betula pendula Roth) provenances in native and non-native photoperiods.

Continuous light (CL) is available throughout the polar day for plants in the Arctic during the growing season, whereas provenances of the same species experience a very different environment with non-CL (NCL) just a few latitudes to the south. Both provenances need to acclimate to climate warming, yet we lack comprehensive understanding of how their growth, photosynthesis and leaf traits differ. Further, the provenances presumably have morphological and physiological adaptations to their native environments and therefore differ in response to photoperiod. We tested the height growth, leaf longevity, biomass accumulation, biomass allocation and rates of gas exchange of northern (67°N) and southern (61°N) Finnish silver birch (Betula pendula Roth) origins in CL- and NCL-treatments in a 4-month chamber experiment. Irrespective of photoperiod, 67°N had higher area-based photosynthetic rate (Anet), stomatal conductance (gs) and relative height growth rate (RGR), but lower stomatal density and fewer branches and leaves than 61°N. Photoperiod affected height growth cessation, biomass and photosynthetic traits, whereas leaf longevity and many leaf functional traits remained unchanged. In CL, both provenances had lower gs, higher RGR, increased shoot:root ratio and increased sink sizes (more branching, more leaves, increased total plant dry weight) compared with NCL. In NCL, 67°N ceased height growth earlier than in CL, which altered biomass accumulation and distribution patterns. Northern conditions impose challenges for plant growth and physiology. Whether a provenance inhabits and is adapted to an area with or without CL can also affect its response to the changing climate. Northern birches may have adapted to CL and the short growing season with a 'polar day syndrome' of traits, including relatively high gas exchange rates with low leaf biomass and growth traits that are mainly limited by the environment and the earlier growth cessation (to avoid frost damage).

Emissions of biogenic volatile organic compounds from adjacent boreal fen and bog as impacted by vegetation composition.

Peatland ecosystems emit biogenic volatile organic compounds (BVOC), which have a net cooling impact on the climate. However, the quality and quantity of BVOC emissions, and how they are regulated by vegetation and peatland type remain poorly understood. Here we measured BVOC emissions with dynamic enclosures from two major boreal peatland types, a minerotrophic fen and an ombrotrophic bog situated in Siikaneva, southern Finland and experimentally assessed the role of vegetation by removing vascular vegetation with or without the moss layer. Our measurements from four campaigns during growing seasons in 2017 and 2018 identified emissions of 59 compounds from nine different chemical groups. Isoprene accounted for 81 % of BVOC emissions. Measurements also revealed uptake of dichloromethane. Total BVOC emissions and the emissions of isoprene, monoterpenoids, sesquiterpenes, homoterpenes, and green leaf volatiles were tightly connected to vascular plants. Isoprene and sesquiterpene emissions were associated with sedges, whereas monoterpenoids and homoterpenes were associated with shrubs. Additionally, isoprene and alkane emissions were higher in the fen than in the bog and they significantly contributed to the higher BVOC emissions from intact vegetation in the fen. During an extreme drought event in 2018, emissions of organic halides were absent. Our results indicate that climate change with an increase in shrub cover and increased frequency of extreme weather events may have a negative impact on total BVOC emissions that otherwise are predicted to increase in warmer temperatures. However, these changes also accompanied a change in BVOC emission quality. As different compounds differ in their capacity to form secondary organic aerosols, the ultimate climate impact of peatland BVOC emissions may be altered.

Metabolite Profiling of Paraquat Tolerant Arabidopsis thaliana Radical-induced Cell Death1 (rcd1)-A Mediator of Antioxidant Defence Mechanisms.

RADICAL-INDUCED CELL DEATH1 (RCD1) is an Arabidopsis thaliana nuclear protein that is disrupted during oxidative stress. RCD1 is considered an important integrative node in development and stress responses, and the rcd1 plants have several phenotypes and altered resistance to a variety of abiotic and biotic stresses. One of the phenotypes of rcd1 is resistance to the herbicide paraquat, but the mechanisms behind it are unknown. Paraquat causes a rapid burst of reactive oxygen species (ROS) initially in the chloroplast. We performed multi-platform metabolomic analyses in wild type Col-0 and paraquat resistant rcd1 plants to identify pathways conveying resistance and the function of RCD1 in this respect. Wild type and rcd1 plants were clearly distinguished by their abundance of antioxidants and specialized metabolites and their responses to paraquat. The lack of response in rcd1 suggested constitutively active defense against ROS via elevated flavonoid, glutathione, ß-carotene, and tocopherol levels, whereas its ascorbic acid levels were compromised under non-stressed control conditions when compared to Col-0. We propose that RCD1 acts as a hub that maintains basal antioxidant system, and its inactivation induces defense responses by enhancing the biosynthesis and redox cycling of low molecular weight antioxidants and specialized metabolites with profound antioxidant activities alleviating oxidative stress.

Mass Spectrometry Imaging of Arabidopsis thaliana Leaves at the Single-Cell Level by Infrared Laser Ablation Atmospheric Pressure Photoionization (LAAPPI).

In this study, we show that infrared laser ablation atmospheric pressure photoionization mass spectrometry (LAAPPI-MS) imaging with 70 µm lateral resolution allows for the analysis of Arabidopsis thaliana (A. thaliana) leaf substructures ranging from single-cell trichomes and the interveinal leaf lamina to primary, secondary, and tertiary veins. The method also showed its potential for depth profiling analysis for the first time by mapping analytes at the different depths of the leaf and spatially resolving the topmost trichomes and cuticular wax layer from the underlying tissues. Negative ion LAAPPI-MS detected many different flavonol glycosides, fatty acids, fatty acid esters, galactolipids, and glycosphingolipids, whose distributions varied significantly between the different substructures of A. thaliana leaves. The results show that LAAPPI-MS provides a highly promising new tool to study the role of metabolites in plants.

Assessment of a portable UV-Vis spectrophotometer's performance in remote areas: Stream water DOC, Fe content and spectral data.

This paper presents data for the assessment of a portable UV-Vis spectrophotometer's performance on predicting stream water DOC and Fe content. The dataset contains DOC and Fe concentrations by laboratory methods, in-situ and ex-situ spectral absorbances, monitoring environmental indexes such as water depth, temperature, turbidity and voltage. The records in Yli-Nuortti river (Cold station, Finland) took place during the hydrological year 2018-2019 and in Krycklan (C4 and C5, Sweden) during the hydrological years 2016-2019. The data analyses were conducted with 'pls' and 'caret' package in R. The correlation coefficient (R), root-mean-square deviation (RMSD), standard deviation (STD) and bias were used to check the performance of the models. This dataset can be combined with datasets from other regions around the world to build more universal models. For discussion and more information of the dataset creation, please refer to the full-length article "Assessment of a portable UV-Vis spectrophotometer's performance for stream water DOC and Fe content monitoring in remote areas" [1].

Assessment of a portable UV-Vis spectrophotometer's performance for stream water DOC and Fe content monitoring in remote areas.

Quantification of dissolved organic carbon (DOC) and iron (Fe) in surface waters is critical for understanding the water quality dynamics, brownification and carbon balance in the northern hemisphere. Especially in the remote areas, sampling and laboratory analysis of DOC and Fe content at a sufficient temporal frequency is difficult. Ultraviolet-visible (UV-Vis) spectrophotometry is a promising tool for water quality monitoring to increase the sampling frequency and applications in remote regions. The aim of this study was (1) to investigate the performance of an in-situ UV-Vis spectrophotometer for detecting spectral absorbances in comparison with a laboratory benchtop instrument; (2) to analyse the stability of DOC and Fe estimates from UV-Vis spectrophotometers among different rivers using multivariate methods; (3) to compare site-specific calibration of models to pooled models and investigate the extrapolation of DOC and Fe predictions from one catchment to another. This study indicates that absorbances that were measured by UV-Vis sensor explained 96% of the absorbance data from the laboratory benchtop instrument. Among the three tested multivariate methods, multiple stepwise regression (MSR) was the best model for both DOC and Fe predictions. Accurate and unbiased models for multiple watersheds for DOC were built successfully, and these models could be extrapolated from one watershed to another even without site-specific calibration for DOC. However, for Fe the combination of different datasets was not possible.

Strategy by latitude? Higher photosynthetic capacity and root mass fraction in northern than southern silver birch (Betula pendula Roth) in uniform growing conditions.

Growth of northern trees is limited by short growing seasons. In multi-year trials, northern trees usually grow less than southern ones but can have higher gas exchange, whereas differences in biomass allocation and its relation to photosynthesis are less known. We characterized silver birch (Betula pendula Roth) provenances from southern (latitude 61°) and northern (latitude 67°) Finland in uniform chamber conditions. In a time-series experiment, we measured traits related to growth, biomass allocation and photosynthesis, and determined gas exchange responses to temperature and light. We found provenance differences in photosynthetic capacity and growth. The northern provenance allocated relatively more to roots, having a higher root mass fraction and lower shoot:root ratio than the southern provenance. On the other hand, the northern provenance had fewer leaves and lower total leaf dry weight (DW) than the southern provenance. The northern provenance attained higher rates of net photosynthesis (Anet) and higher stomatal conductance (gs) in all measured temperatures and higher photosynthesis at the optimum temperature (Aopt) than the southern provenance, but there was no difference in the optimum temperature of photosynthesis (Topt, 18.3 °C for the southern provenance vs 18.9 °C for the northern one). Photosynthetic light response curves showed no between-provenance differences. In a time-series, the northern provenance had higher Anet than the southern provenance, but gs was similar. The northern provenance had higher maximum quantum yield of photosystem II photochemistry (Fv/Fm) than the southern provenance. There were no differences between provenances in height, total plant DW, shoot DW, root DW or shoot mass fraction. Our results suggest that the provenances occupy a common thermal niche, or can at least relatively quickly acclimate to a common growth temperature. Thus, carbon assimilation of these northern trees may not be significantly affected by rising temperatures alone. In an equal photoperiod and optimal conditions, we found different one-season biomass accumulation strategies: southern trees grow with more leaves, while northern trees reach similar total assimilation (total DW, height) with more efficient photosynthetic capacity per leaf area (higher gas exchange, higher Fv/Fm) and relatively more investment in the below-ground fraction of the plant.

Retene, pyrene and phenanthrene cause distinct molecular-level changes in the cardiac tissue of rainbow trout (Oncorhynchus mykiss) larvae, part 2 - Proteomics and metabolomics.

Polycyclic aromatic hydrocarbons (PAHs) are global contaminants of concern. Despite several decades of research, their mechanisms of toxicity are not very well understood. Early life stages of fish are particularly sensitive with the developing cardiac tissue being a main target of PAHs toxicity. The mechanisms of cardiotoxicity of the three widespread model polycyclic aromatic hydrocarbons (PAHs) retene, pyrene and phenanthrene were explored in rainbow trout (Oncorhynchus mykiss) early life stages. Newly hatched larvae were exposed to sublethal doses of each individual PAH causing no detectable morphometric alterations. Changes in the cardiac proteome and metabolome were assessed after 7 or 14 days of exposure to each PAH. Phase I and II enzymes regulated by the aryl hydrocarbon receptor were significantly induced by all PAHs, with retene being the most potent compound. Retene significantly altered the level of several proteins involved in key cardiac functions such as muscle contraction, cellular tight junctions or calcium homeostasis. Those findings were quite consistent with previous reports regarding the effects of retene on the cardiac transcriptome. Significant changes in proteins linked to iron and heme metabolism were observed following exposure to pyrene. While phenanthrene also altered the levels of several proteins in the cardiac tissue, no clear mechanisms or pathways could be highlighted. Due to high variability between samples, very few significant changes were detected in the cardiac metabolome overall. Slight but significant changes were still observed for pyrene and phenanthrene, suggesting possible effects on several energetic or signaling pathways. This study shows that early exposure to different PAHs can alter the expression of key proteins involved in the cardiac function, which could potentially affect negatively the fitness of the larvae and later of the juvenile fish.

Primary Metabolite Responses to Oxidative Stress in Early-Senescing and Paraquat Resistant Arabidopsis thaliana rcd1 (Radical-Induced Cell Death1).

Rcd1 (radical-induced cell death1) is an Arabidopsis thaliana mutant, which exhibits high tolerance to paraquat [methyl viologen (MV)], herbicide that interrupts photosynthetic electron transport chain causing the formation of superoxide and inhibiting NADPH production in the chloroplast. To understand the biochemical mechanisms of MV-resistance and the role of RCD1 in oxidative stress responses, we performed metabolite profiling of wild type (Col-0) and rcd1 plants in light, after MV exposure and after prolonged darkness. The function of RCD1 has been extensively studied at transcriptomic and biochemical level, but comprehensive metabolite profiling of rcd1 mutant has not been conducted until now. The mutant plants exhibited very different metabolic features from the wild type under light conditions implying enhanced glycolytic activity, altered nitrogen and nucleotide metabolism. In light conditions, superoxide production was elevated in rcd1, but no metabolic markers of oxidative stress were detected. Elevated senescence-associated metabolite marker levels in rcd1 at early developmental stage were in line with its early-senescing phenotype and possible mitochondrial dysfunction. After MV exposure, a marked decline in the levels of glycolytic and TCA cycle intermediates in Col-0 suggested severe plastidic oxidative stress and inhibition of photosynthesis and respiration, whereas in rcd1 the results indicated sustained photosynthesis and respiration and induction of energy salvaging pathways. The accumulation of oxidative stress markers in both plant lines indicated that MV-resistance in rcd1 derived from the altered regulation of cellular metabolism and not from the restricted delivery of MV into the cells or chloroplasts. Considering the evidence from metabolomic, transcriptomic and biochemical studies, we propose that RCD1 has a negative effect on reductive metabolism and rerouting of the energy production pathways. Thus, the altered, highly active reductive metabolism, energy salvaging pathways and redox transfer between cellular compartments in rcd1 could be sufficient to avoid the negative effects of MV-induced toxicity.

Differences in growth and gas exchange between southern and northern provenances of silver birch (Betula pendula Roth) in northern Europe.

Due to its ubiquity across northern latitudes, silver birch (Betula pendula Roth) is an attractive model species for studying geographical trait variation and acclimation capacity. Six birch provenances from 60 to 67°N across Finland were grown in a common garden and studied for provenance and genotype variation. We looked for differences in height growth, photosynthetic gas exchange and chlorophyll content index (CCI) and compared the gas exchange of early and late leaves on short and long shoots, respectively. The provenances stratified into southern and northern groups. Northern provenances attained less height growth increment and had higher stomatal conductance (gs) and lower intrinsic water-use efficiency (WUE, Anet/gs) than southern provenances, whereas net photosynthesis (Anet) or CCI did not show clear grouping. Short shoot leaves had lower gs and higher WUE than long shoot leaves in all provenances, but there was no difference in Anet between shoot types. The separation of the provenances into two groups according to their physiological responses might reflect the evolutionary history of B. pendula. Latitudinal differences in gas exchange and water use traits can have plausible consequences for global carbon and water fluxes in a warming climate.

Narrowband-autofluorescence imaging for bone analysis.

We present a new autofluorescence-imaging method for bone analysis. This method, based on the autofluorescence of bone, provides color images in microscopic scale. The color images are created from three monochrome images acquired with optimal excitation- and emission-wavelengths combinations. The choice of these combinations were determined from the study of two-dimensional distributions of bone-features-bispectral autofluorescence in the visible- and ultraviolet-spectral range. We demonstrate that main-bone features visualized with MG-staining method can also be visualized in the autofluorescence-color image. Furthermore, the autofluorescence-color image presents features hardly distinguished in a histological-bone section.

Arabidopsis RCD1 coordinates chloroplast and mitochondrial functions through interaction with ANAC transcription factors.

Reactive oxygen species (ROS)-dependent signaling pathways from chloroplasts and mitochondria merge at the nuclear protein RADICAL-INDUCED CELL DEATH1 (RCD1). RCD1 interacts in vivo and suppresses the activity of the transcription factors ANAC013 and ANAC017, which mediate a ROS-related retrograde signal originating from mitochondrial complex III. Inactivation of RCD1 leads to increased expression of mitochondrial dysfunction stimulon (MDS) genes regulated by ANAC013 and ANAC017. Accumulating MDS gene products, including alternative oxidases (AOXs), affect redox status of the chloroplasts, leading to changes in chloroplast ROS processing and increased protection of photosynthetic apparatus. ROS alter the abundance, thiol redox state and oligomerization of the RCD1 protein in vivo, providing feedback control on its function. RCD1-dependent regulation is linked to chloroplast signaling by 3'-phosphoadenosine 5'-phosphate (PAP). Thus, RCD1 integrates organellar signaling from chloroplasts and mitochondria to establish transcriptional control over the metabolic processes in both organelles.

Transcriptional effects of cadmium on iron homeostasis differ in calamine accessions of Noccaea caerulescens.

Calamine accessions of the zinc/cadmium/nickel hyperaccumulator, Noccaea caerulescens, exhibit striking variation in foliar cadmium accumulation in nature. The Ganges accession (GA) from Southern France displays foliar cadmium hyperaccumulation (>1000 µg g-1 DW), whereas the accession La Calamine (LC) from Belgium, with similar local soil metal composition, does not (<100 µg g-1 DW). All calamine accessions are cadmium hypertolerant. To find out the differences between LC and GA in their basic adaptation mechanisms, we bypassed the cadmium excluding phenotype of LC by exposing the plants to 50 µm cadmium in hydroponics, achieving equal cadmium accumulation in the shoots. The iron content increased in the roots of both accessions. GA exhibited significant decreases in manganese and zinc contents in the roots and shoots, approaching those in LC. Altogether 702 genes responded differently to cadmium exposure between the accessions, 157 and 545 in the roots and shoots, respectively. Cadmium-exposed LC showed a stress response and had decreased levels of a wide range of photosynthesis-related transcripts. GA showed less changes, mainly exhibiting an iron deficiency-like response. This included increased expression of genes encoding five iron deficiency-regulated bHLH transcription factors, ferric reduction oxidase FRO2, iron transporters IRT1 and OPT3, and nicotianamine synthase NAS1, and decreased expression of genes encoding ferritins and NEET (a NEET family iron-sulfur protein), which is possibly involved in iron transfer, distribution and/or management. The function of the IRT1 gene in the accessions was compared. We conclude that the major difference between the two accessions is in the way they cope with iron under cadmium exposure.

Intrapopulation Genotypic Variation of Foliar Secondary Chemistry during Leaf Senescence and Litter Decomposition in Silver Birch (Betula pendula).

Abundant secondary metabolites, such as condensed tannins, and their interpopulation genotypic variation can remain through plant leaf senescence and affect litter decomposition. Whether the intrapopulation genotypic variation of a more diverse assortment of secondary metabolites equally persists through leaf senescence and litter decomposition is not well understood. We analyzed concentrations of intracellular phenolics, epicuticular flavonoid aglycones, epicuticular triterpenoids, condensed tannins, and lignin in green leaves, senescent leaves and partly decomposed litter of silver birch, Betula pendula. Broad-sense heritability (H2) and coefficient of genotypic variation (CVG) were estimated for metabolites in senescent leaves and litter using 19 genotypes selected from a B. pendula population in southern Finland. We found that most of the secondary metabolites remained through senescence and decomposition and that their persistence was related to their chemical properties. Intrapopulation H2 and CVG for intracellular phenolics, epicuticular flavonoid aglycones and condensed tannins were high and remarkably, increased from senescent leaves to decomposed litter. The rank of genotypes in metabolite concentrations was persistent through litter decomposition. Lignin was an exception, however, with a diminishing genotypic variation during decomposition, and the concentrations of lignin and condensed tannins had a negative genotypic correlation in the senescent leaves. Our results show that secondary metabolites and their intrapopulation genotypic variation can for the most part remain through leaf senescence and early decomposition, which is a prerequisite for initial litter quality to predict variation in litter decomposition rates. Persistent genotypic variation also opens an avenue for selection to impact litter decomposition in B. pendula populations through acting on their green foliage secondary chemistry. The negative genotypic correlations and diminishing heritability of lignin concentrations may, however, counteract this process.

Low vapor pressure deficit reduces glandular trichome density and modifies the chemical composition of cuticular waxes in silver birch leaves.

Cuticular wax layer is the first barrier against the outside environment and the first defense encountered by herbivores and pathogens. The effects of environmental factors on cuticular chemistry, and on the formation of glandular trichomes that account for the storage and secretion of lipophilic compounds to the leaf surface are poorly understood. Low vapor pressure deficit (VPD) has shown to reduce the nitrogen (N) status of plants. Thus, we studied the effects of elevated air humidity, indicated as VPD, and the effect of N fertilization on cuticular waxes and glandular trichome density in silver birch (Betula pendula Roth). Experiments were carried out in growth chambers with juvenile plants and in a long-term field experiment with older trees. Low VPD reduced the glandular trichome density in both experiments, in chamber and in field. The contents of the major triterpenoid and flavonoid aglycones correlated positively with glandular trichome density, which supports the role of trichomes in the exudation of secondary compounds to the leaf surface. A closer examination of the cuticular wax chemistry in the chamber experiment revealed that low VPD and N supply affected the composition of cuticular waxes, but not the total wax content. The deposition of different wax compounds followed a co-ordinated pattern in birch leaves, but different compound groups varied in their responses to N fertilization and low VPD. Low VPD reduced the hydrophobicity of cuticular waxes, as demonstrated by lower alkane content and less hydrophobic flavonoid profile in low VPD than in high VPD. Reduced hydrophobicity of the wax layer is presumed to increase leaf wettability. Together with reduced trichome density in low VPD it may enhance the susceptibility of trees to fungal pathogens and herbivores. High N supply under low VPD reduced the effect of low VPD on the cuticular wax composition. Total fatty acid content and the expression of ß-amyrin synthase were lower under high N supply than under moderate N supply irrespective of VPD treatment. Nitrogen availability and decreasing VPD will modify leaf surface properties in silver birch and thereby affect tree defence against abiotic and biotic stress factors that emerge under climate change.

Artificially decreased vapour pressure deficit in field conditions modifies foliar metabolite profiles in birch and aspen.

Relative air humidity (RH) is expected to increase in northern Europe due to climate change. Increasing RH reduces the difference of water vapour pressure deficit (VPD) between the leaf and the atmosphere, and affects the gas exchange of plants. Little is known about the effects of decreased VPD on plant metabolism, especially under field conditions. This study was conducted to determine the effects of artificially decreased VPD on silver birch (Betula pendula Roth.) and hybrid aspen (Populus tremula L.×P. tremuloides Michx.) foliar metabolite and nutrient profiles in a unique free air humidity manipulation (FAHM) field experiment during the fourth season of humidity manipulation, in 2011. Long-term exposure to decreased VPD modified nutrient homeostasis in tree leaves, as demonstrated by a lower N concentration and N:P ratio in aspen leaves, and higher Na concentration and lower K:Na ratio in the leaves of both species in decreased VPD than in ambient VPD. Decreased VPD caused a shift in foliar metabolite profiles of both species, affecting primary and secondary metabolites. Metabolic adjustment to decreased VPD included elevated levels of starch and heptulose sugars, sorbitol, hemiterpenoid and phenolic glycosides, and α-tocopherol. High levels of carbon reserves, phenolic compounds, and antioxidants under decreased VPD may modify plant resistance to environmental stresses emerging under changing climate.

Low vapour pressure deficit affects nitrogen nutrition and foliar metabolites in silver birch.

Air humidity indicated as vapour pressure deficit (VPD) is directly related to transpiration and stomatal function of plants. We studied the effects of VPD and nitrogen (N) supply on leaf metabolites, plant growth, and mineral nutrition with young micropropagated silver birches (Betula pendula Roth.) in a growth chamber experiment. Plants that were grown under low VPD for 26 d had higher biomass, larger stem diameter, more leaves, fewer fallen leaves, and larger total leaf area than plants that were grown under high VPD. Initially, low VPD increased height growth rate and stomatal conductance; however, the effect was transient and the differences between low and high VPD plants became smaller with time. Metabolic adjustment to low VPD reflected N deficiency. The concentrations of N, iron, chlorophyll, amino acids, and soluble carbohydrates were lower and the levels of starch, quercetin glycosides, and raffinose were higher in the leaves that had developed under low VPD compared with high VPD. Additional N supply did not fully overcome the negative effect of low VPD on nutrient status but it diminished the effects of low VPD on leaf metabolism. Thus, with high N supply, the glutamine to glutamate ratio and starch production under low VPD became comparable with the levels under high VPD. The present study demonstrates that low VPD affects carbon and nutrient homeostasis and modifies N allocation of plants.