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Biogenic secondary organic aerosols (SOAs) can be formed from the oxidation of plant volatiles in the atmosphere. Herbivore-induced plant volatiles (HIPVs) can elicit plant defenses, but whether such ecological functions persist after they form SOAs was previously unknown. Here we show that Scots pine seedlings damaged by large pine weevils feeding on their roots release HIPVs that trigger defenses in neighboring conspecific plants. The biological activity persisted after HIPVs had been oxidized to form SOAs, which was indicated by receivers displaying enhanced photosynthesis, primed volatile defenses, and reduced weevil damage. The elemental composition and quantity of SOAs likely determines their biological functions. This work demonstrates that plant-derived SOAs can mediate interactions between plants, highlighting their ecological significance in ecosystems.

Forests play a crucial role in global carbon cycling by absorbing and storing significant amounts of atmospheric carbon dioxide. Although boreal forests contribute to approximately 45% of the total forest carbon sink, tree growth and soil carbon sequestration are constrained by nutrient availability. Here, we examine if long-term nutrient input enhances tree productivity and whether this leads to carbon storage or whether stimulated microbial decomposition of organic matter limits soil carbon accumulation. Over six decades, nitrogen, phosphorus, and calcium were supplied to a Pinus sylvestris-dominated boreal forest. We found that nitrogen fertilization alone or together with calcium and/or phosphorus increased tree biomass production by 50% and soil carbon sequestration by 65% compared to unfertilized plots. However, the nonlinear relationship observed between tree productivity and soil carbon stock across treatments suggests microbial regulation. When phosphorus was co-applied with nitrogen, it acidified the soil, increased fungal biomass, altered microbial community composition, and enhanced biopolymer degradation capabilities. While no evidence of competition between ectomycorrhizal and saprotrophic fungi has been observed, key functional groups with the potential to reduce carbon stocks were identified. In contrast, when nitrogen was added without phosphorus, it increased soil carbon sequestration because microbial activity was likely limited by phosphorus availability. In conclusion, the addition of nitrogen to boreal forests may contribute to global warming mitigation, but this effect is context dependent., (© 2024 The Author(s). Global Change Biology published by John Wiley & Sons Ltd.)

Understanding the dynamics of δ 13 C and δ 18 O in modern resin is crucial for interpreting (sub)fossilized resin records and resin production dynamics. We measured the δ 13 C and δ 18 O 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 δ 13 C and δ 18 O at an intra-seasonal scale. Resin δ 13 C was c. 2‰ lower than sucrose δ 13 C, in both twigs and stems, likely due to the loss of 13 C-enriched C-1 atoms of pyruvate during isoprene formation and kinetic isotope effects during diterpene synthesis. Resin δ 18 O was c. 20‰ higher than xylem water δ 18 O and c. 20‰ lower than δ 18 O of water-soluble carbohydrates, possibly caused by discrimination against 18 O during O 2 -based diterpene oxidation and 35%-50% oxygen atom exchange with water. Resin δ 13 C and δ 18 O 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 δ 13 C and δ 18 O as a long-term proxy for physiological and environmental changes., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Stem CO2 efflux is an important component of the carbon balance in forests. The efflux is considered to principally reflect the net result of two dominating and opposing processes: stem respiration and stem photosynthesis. In addition, transport of CO2 in xylem sap is thought to play an appreciable role in affecting the net flux. This work presents an approach to partition stem CO2 efflux among these processes using sap-flux data and CO2-exchange measurements from dark and transparent chambers placed on mature Scots pine (Pinus sylvestris) trees. Seasonal changes and monthly parameters describing the studied processes were determined. Respiration contributed most to stem net CO2 flux, reaching up to 79% (considering the sum of the absolute values of stem respiration, stem photosynthesis, and flux from CO2 transported in xylem sap to be 100%) in June, when stem growth was greatest. The contribution of photosynthesis accounted for up to 13% of the stem net CO2 flux, increasing over the monitoring period. CO2 transported axially with sap flow decreased towards the end of the growing season. At a reference temperature, respiration decreased starting around midsummer, while its temperature sensitivity increased during the summer. A decline was observed for photosynthetic quantum yield around midsummer together with a decrease in light-saturation point. The proposed approach facilitates modeling net stem CO2 flux at a range of time scales., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

The photosynthetic acclimation of boreal evergreen conifers is controlled by regulatory and photoprotective mechanisms that allow conifers to cope with extreme environmental changes. However, the underlying dynamics of photosystem II (PSII) and photosystem I (PSI) remain unresolved. Here, we investigated the dynamics of PSII and PSI during the spring recovery of photosynthesis in Pinus sylvestris and Picea abies using a combination of chlorophyll a fluorescence, P700 difference absorbance measurements, and quantification of key thylakoid protein abundances. In particular, we derived a new set of PSI quantum yield equations, correcting for the effects of PSI photoinhibition. Using the corrected equations, we found that the seasonal dynamics of PSII and PSI photochemical yields remained largely in balance, despite substantial seasonal changes in the stoichiometry of PSII and PSI core complexes driven by PSI photoinhibition. Similarly, the previously reported seasonal up-regulation of cyclic electron flow was no longer evident, after accounting for PSI photoinhibition. Overall, our results emphasize the importance of considering the dynamics of PSII and PSI to elucidate the seasonal acclimation of photosynthesis in overwintering evergreens. Beyond the scope of conifers, our corrected PSI quantum yields expand the toolkit for future studies aimed at elucidating the dynamic regulation of PSI., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

Shoot-level emissions of aerobically produced methane (CH 4 ) may be an overlooked source of tree-derived CH 4 , but insufficient understanding of the interactions between their environmental and physiological drivers still prevents the reliable upscaling of canopy CH 4 fluxes. We utilised a novel automated chamber system to continuously measure CH 4 fluxes from the shoots of Pinus sylvestris (Scots pine) saplings under drought to investigate how canopy CH 4 fluxes respond to the drought-induced alterations in their physiological processes and to isolate the shoot-level production of CH 4 from soil-derived transport and photosynthesis. We found that aerobic CH 4 emissions are not affected by the drought-induced stress, changes in physiological processes, or decrease in photosynthesis. Instead, these emissions vary on short temporal scales with environmental drivers such as temperature, suggesting that they result from abiotic degradation of plant compounds. Our study shows that aerobic CH 4 emissions from foliage are distinct from photosynthesis-related processes. Thus, instead of photosynthesis rates, it is more reliable to construct regional and global estimates for the aerobic CH 4 emission based on regional differences in foliage biomass and climate, also accounting for short-term variations of weather variables such as air temperature and solar radiation., (© 2024 The Authors New Phytologist © 2024 New Phytologist Foundation.)

Tree growth in boreal forests is driven by ectomycorrhizal fungal mobilisation of organic nitrogen and mineral nutrients in soils with discrete organic and mineral horizons. However, there are no studies of how ectomycorrhizal mineral weathering and organic nitrogen mobilisation processes are integrated across the soil profile. We studied effects of organic matter (OM) availability on ectomycorrhizal functioning by altering the proportions of natural organic and mineral soil in reconstructed podzol profiles containing Pinus sylvestris plants, using 13 CO 2 pulse labelling, patterns of naturally occurring stable isotopes ( 26 Mg and 15 N) and high-throughput DNA sequencing of fungal amplicons. Reduction in OM resulted in nitrogen limitation of plant growth and decreased allocation of photosynthetically derived carbon and mycelial growth in mineral horizons. Fractionation patterns of 26 Mg indicated that magnesium mobilisation and uptake occurred primarily in the deeper mineral horizon and was driven by carbon allocation to ectomycorrhizal mycelium. In this horizon, relative abundance of ectomycorrhizal fungi, carbon allocation and base cation mobilisation all increased with increased OM availability. Allocation of carbon through ectomycorrhizal fungi integrates organic nitrogen mobilisation and mineral weathering across soil horizons, improving the efficiency of plant nutrient acquisition. Our findings have fundamental implications for sustainable forest management and belowground carbon sequestration., (© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.)

The loss of leaves and needles in tree crowns and tree mortality are increasing worldwide, mostly as a result of more frequent and severe drought stress. Scots pine (Pinus sylvestris L.) is a tree species that is strongly affected by these developments in many regions of Europe and Asia. So far, changes in metabolic pathways and metabolite profiles in needles and roots on the trajectory toward mortality are unknown, although they could contribute to a better understanding of the mortality mechanisms. Therefore, we linked long-term observations of canopy defoliation and tree mortality with the characterization of the primary metabolite profile in needles and fine roots of Scots pines from a forest site in the Swiss Rhone valley. Our results show that Scots pines are able to maintain metabolic homeostasis in needles over a wide range of canopy defoliation levels. However, there is a metabolic tipping point at around 80-85% needle loss. Above this threshold, many stress-related metabolites (particularly osmoprotectants, defense compounds and antioxidants) increase in the needles, whereas they decrease in the fine roots. If this defoliation tipping point is exceeded, the trees are very likely to die within a few years. The different patterns between needles and roots indicate that mainly belowground carbon starvation impairs key functions for tree survival and suggest that this is an important factor explaining the increasing mortality of Scots pines., (© The Author(s) 2024. Published by Oxford University Press.)

Carbon isotope composition of tree-ring (δ 13 C Ring ) is a commonly used proxy for environmental change and ecophysiology. δ 13 C Ring reconstructions are based on a solid knowledge of isotope fractionations during formation of primary photosynthates (δ 13 C P ), such as sucrose. However, δ 13 C Ring is not merely a record of δ 13 C P . Isotope fractionation processes, which are not yet fully understood, modify δ 13 C P during sucrose transport. We traced, how the environmental intra-seasonal δ 13 C P signal changes from leaves to phloem, tree-ring and roots, for 7 year old Pinus sylvestris, using δ 13 C analysis of individual carbohydrates, δ 13 C Ring laser ablation, leaf gas exchange and enzyme activity measurements. The intra-seasonal δ 13 C P dynamics was clearly reflected by δ 13 C Ring , suggesting negligible impact of reserve use on δ 13 C Ring . However, δ 13 C P became increasingly 13 C-enriched during down-stem transport, probably due to post-photosynthetic fractionations such as sink organ catabolism. In contrast, δ 13 C of water-soluble carbohydrates, analysed for the same extracts, did not reflect the same isotope dynamics and fractionations as δ 13 C P , but recorded intra-seasonal δ 13 C P variability. The impact of environmental signals on δ 13 C Ring , and the 0.5 and 1.7‰ depletion in photosynthates compared ring organic matter and tree-ring cellulose, respectively, are useful pieces of information for studies exploiting δ 13 C Ring ., (© 2023 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Forest trees are subjected to multiple stressors during their long lifetime and therefore require effective and finely regulated stress-protective systems. Stressors can induce protective systems either directly or with the involvement of stress memory mechanisms. Stress memory has only begun to be uncovered in model plants and is unexplored in coniferous species. Therefore, we studied the possible role of stress memory in the regulation of the accumulation of stress-protective compounds (heat shock proteins, dehydrins, proline) in the needles of naturally grown Scots pine and Norway spruce trees subjected to the subsequent action of long-term (multiyear) and short-term (seasonal) water shortages. Although the water deficit was relatively mild, it significantly influenced the pattern of expression of stress memory-related heat shock factor (HSF) and SWI/SNF genes, indicating the formation of stress memory in both species. In spruce, dehydrin accumulation was increased by water shortage in a manner compatible with Type II stress memory. The accumulation of HSP40 in spruce needles was positively influenced by long-term water shortage, but this increase was unlikely to be of biological importance due to the concomitant decrease in HSP70, HSP90 and HSP101 accumulation. Finally, proline accumulation was negatively influenced by short-term water deficit in spruce. In pine, no one protective compound accumulated in response to water stress. Taken together, the results indicate that the accumulation of stress-protective compounds was generally independent of stress memory effects both in pine and in spruce., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Green organisms evolve oxygen (O 2 ) via photosynthesis and consume it by respiration. Generally, net O 2 consumption only becomes dominant when photosynthesis is suppressed at night. Here, we show that green thylakoid membranes of Scots pine (Pinus sylvestris L) and Norway spruce (Picea abies) needles display strong O 2 consumption even in the presence of light when extremely low temperatures coincide with high solar irradiation during early spring (ES). By employing different electron transport chain inhibitors, we show that this unusual light-induced O 2 consumption occurs around photosystem (PS) I and correlates with higher abundance of flavodiiron (Flv) A protein in ES thylakoids. With P700 absorption changes, we demonstrate that electron scavenging from the acceptor-side of PSI via O 2 photoreduction is a major alternative pathway in ES. This photoprotection mechanism in vascular plants indicates that conifers have developed an adaptative evolution trajectory for growing in harsh environments., (© 2023. The Author(s).)

The effects of Cu, Ni, and Cd on the Pinus sylvestris metabolome was studied in experimental conditions by gas chromatography-mass spectrometry (GC-MS). Structural changes in plant metabolite network became detectable on day 6 of exposure to the metals, 3-6 days earlier than visual signs of toxicity developed. Differences at the metabolome level arose earlier in a control group of plants, and specific effects of particular metals on the plant metabolome became distinct on day 9. Both nature and concentration of a metal equally contributed to the plant metabolome clustering. Plant responses (changes in concentrations of individual metabolites) to metal exposure substantially differed depending on the metal concentration (1 or 5 mM) and nature. The effects of Cd and Cu were generally similar, while the effect of Ni was often different. Dynamic changes visualized in plant metabolite matrix reflected the changes in its correlation structure, rather than depending on the set of particular compounds., (© 2022. Pleiades Publishing, Ltd.)

Manganese deficiency is a serious plant nutritional disorder, resulting in the loss of crop productivity in many parts of the world. Despite the progress made in the study of angiosperms, the demand for Mn in gymnosperms and the physiological responses to Mn deficiency remain unexplored. We studied the influence of Mn deficiency for 24 weeks on Pinus sylvestris L. seedling growth, ion homeostasis, pigment contents, lipid peroxidation, chlorophyll fluorescence indices and the transcript levels of photosynthetic genes and genes involved in chlorophyll biosynthesis. It was shown that Mn-deficient plants demonstrated suppressed growth when the Mn content in the needles decreased below 0.34 µmol/g DW. The contents of photosynthetic pigments decreased when the Mn content in the needles reached 0.10 µmol/g DW. Mn deficiency per se did not lead to a decrease in the nutrient content in the organs of seedlings. Photoinhibition of PSII was observed in Mn-deficient plants, although this was not accompanied by the development of oxidative stress. Mn-deficient plants had an increased transcript abundance of genes ( psbO , psbP , psbQ , psbA and psbC ), encoding proteins directly associated with the Mn cluster also as other proteins involved in photosynthesis, whose activities do not depend on Mn directly. Furthermore, the transcript levels of the genes encoding the large subunit of Rubisco, light-dependent NADPH-protochlorophyllide oxidoreductase and subunits of light-independent protochlorophyllide reductase were also increased in Mn-deficient plants.

Conifers are dominant tree species in boreal forests, but are susceptible to attack by bark beetles. Upon bark beetle attack, conifers release substantial quantities of volatile organic compounds known as herbivore-induced plant volatiles (HIPVs). Earlier studies of broadleaved plants have shown that HIPVs provide information to neighbouring plants, which may enhance their defences. However, the defence responses of HIPV-receiver plants have not been described for conifers. Here we advance knowledge of plant-plant communication in conifers by documenting a suite of receiver-plant responses to bark-feeding-induced volatiles. Scots pine seedlings exposed to HIPVs were more resistant to subsequent weevil feeding and received less damage. Receiver plants had both induced and primed volatile emissions and their resin ducts had an increased epithelial cell (EC) mean area and an increased number of cells located in the second EC layer. Importantly, HIPV exposure increased stomatal conductance and net photosynthesis rate of receiver plants. Receiver-plant responses were also examined under elevated ozone conditions and found to be significantly altered. However, the final defence outcome was not affected. These findings demonstrate that HIPVs modulate conifer metabolism through responses spanning photosynthesis and chemical defence. The responses are adjusted under ozone stress, but the defence benefits remain intact.

Plant defensins demonstrate high structural stability at extreme temperatures and pH values and, in general, are non-toxic to mammalian cells. These properties make them attractive candidates for use in biotechnology and biomedicine. Knowing the structure-function relationship is desirable to guide the design of plant defensin-based applications. Thus far, the broad range of biological activities was described only for one defensin from gymnosperms, the defensin PsDef1 from Scots pine. Here, we report that closely related defensin from the same taxonomy group, PsDef2, differing from PsDef1 by six amino acids, also possesses antimicrobial, antibacterial, and insect α-amylase inhibitory activities. We also report the solution structure and dynamics properties of PsDef2 assessed using a combination of experimental nuclear magnetic resonance (NMR) techniques. Lastly, we perform a comparative analysis of PsDef2 and PsDef1 gaining a molecular-level insight into their structure-dynamics-function relationship., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Introduction: A comprehensive study of enzymes of the antioxidant system (AOS) and phenolic metabolism is an actual subject of biochemical research; changes in the activity of these enzymes can be used as a diagnostic sign. At the same time, practically little attention has been paid to describing the regularities of these enzymatic reactions. The article presents the chemical kinetics study of reactions catalyzed by superoxide dismutase, catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase in Scots pine trunk tissues (Pinus sylvestris L.). The dependence of the enzyme reaction rate on the enzyme concentration and the substrate concentration is presented, and the pH-optimum for each reaction is established., Background: Determination of AOS enzyme activity and PAL activity in woody plants has many difficulties. The chemical composition of pine trunk tissues affects determining AOS enzyme activity and PAL activity. Spectrophotometric determination of AOS enzyme activity and PAL activity gives perfect results when considering all additional controls by taking into account minor characteristics., Objective: This study aimed at determining the AOS enzyme activity in 40-year-old Scots pine (Pinus sylvestris L.) plants growing in the Karelian (Russia) forest seed plantation., Methods: Plant tissues were ground in liquid nitrogen to a uniform mass and homogenized at 4 °C in the buffer containing 50 mM HEPES (pH 7.5), 1 mM EDTA, 1 mM EGTA, 3 mM DTT, 5 mM MgCl2, and 0.5 mM PMSF. After 15-min of extraction, the homogenate was centrifuged at 12000 g for 10 min (MPW-351R centrifuge, Poland). The supernatant was purified on 20 cm3 columns with Sephadex G-250. Aliquots with the highest protein amount were collected. In tissues, the protein concentration was 10-50 μg/ml. Proteins in the extracts were quantified by a Bradford assay. The enzyme activity was determined spectrophotometrically on a SpectroStar Nano plate spectrophotometer (BMG Labtech, Germany)., Results: Our study made it possible to modify the methods for determining the activity of superoxide dismutase, catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase in Scots pine trunk tissues. The enzymatic reaction rate dependence on the enzyme concentration and the substrate concentration was determined, and pH-optimum was also noted. This methodological article also provides formulas for calculating the activities of the enzymes., Conclusion: We found that determining AOS enzyme activity and PAL activity in woody plants is challenging. The chemical composition of the xylem and phloem of pine affects determining AOS enzyme activity and PAL activity. Spectrophotometric determination of AOS enzyme activity and PAL activity gives perfect results when considering all additional controls by taking into account minor characteristics., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The widespread ascomycetous fungus Diplodia pinea is a latent, necrotrophic pathogen in Pinus species causing severe damages and world-wide economic losses. However, the interactions between pine hosts and virulent D. pinea are largely not understood. In the present study, systemic defence responses were investigated in non-inoculated, asymptomatic needles and roots of D. pinea infected saplings of two P. sylvestris provenances under controlled greenhouse conditions. Here, we show that D. pinea infection induced a multitude of systemic responses of the phytohormone profiles and metabolic traits. Shared systemic responses of both pine provenances in needles and roots included increased abscisic acid and jasmonic acid levels. Exclusively in the roots of both provenances, enhanced salicylic acid and reduced indole-3-acetic acid levels, structural biomass, and elevated activities of anti-oxidative enzymes were observed. Despite these similarities, the two pine provenances investigated different significantly in the systemic responses of both, phytohormone profiles and metabolic traits in needles and roots. However, the different systemic responses did not prevent subsequent destruction of non-inoculated needles, but rather prevented damage to the roots. Our results provide a detailed view on systemic defence mechanisms of pine hosts that are of particular significance for the selection of provenances with improved defence capacity., (© 2021 John Wiley & Sons Ltd.)

Polyamines (PA) have a protective role in maintaining growth and development in Scots pine during abiotic stresses. In the present study, a controlled liquid Scots pine embryogenic cell culture was used for studying the responses of PA metabolism related to potassium deficiency. The transcription level regulation of PA metabolism led to the accumulation of putrescine (Put). Arginine decarboxylase ( ADC ) had an increased expression trend under potassium deficiency, whereas spermidine synthase ( SPDS ) expression decreased. Generally, free spermidine (Spd) and spermine (Spm)/ thermospermine (t-Spm) contents were kept relatively stable, mostly by the downregulation of polyamine oxidase (PAO ) expression. The low potassium contents in the culture medium decreased the potassium content of the cells, which inhibited cell mass growth, but did not affect cell viability. The reduced growth was probably caused by repressed metabolic activity and cell division, whereas there were no signs of H 2 O 2 -induced oxidative stress or increased cell death. The low intracellular content of K + decreased the content of Na + . The decrease in the pH of the culture medium indicated that H + ions were pumped out of the cells. Altogether, our findings emphasize the specific role(s) of Put under potassium deficiency and strict developmental regulation of PA metabolism in Scots pine.

Comparative transcriptome analysis provides a useful tool for the exploration of plant-pathogen interaction by allowing in-depth comparison of gene expression between unaffected, inoculated and wounded organisms. Here we present the results of comparative transcriptome analysis in genetically identical one-year-old Scots pine ramets after wounding and inoculation with Heterobasidion annosum . We identified 230 genes that were more than 2-fold upregulated in inoculated samples (compared to controls) and 116 downregulated genes. Comparison of inoculated samp les with wounded samples identified 32 differentially expressed genes (30 were upregulated after inoculation). Several of the genes upregulated after inoculation are involved in protection from oxidative stress, while genes involved in photosynthesis, water transport and drought stress tolerance were downregulated. An NRT3 family protein was the most upregulated transcript in response to both inoculation and wounding, while a U-box domain-containing protein gene was the most upregulated gene comparing inoculation to wounding. The observed transcriptome dynamics suggest involvement of auxin, ethylene, jasmonate, gibberellin and reactive oxygen species pathways and cell wall modification regulation in response to H. annosum infection. The results are compared to methyl jasmonate induced transcriptome dynamics.

Evergreen conifers in boreal forests can survive extremely cold (freezing) temperatures during long dark winter and fully recover during summer. A phenomenon called "sustained quenching" putatively provides photoprotection and enables their survival, but its precise molecular and physiological mechanisms are not understood. To unveil them, here we have analyzed seasonal adjustment of the photosynthetic machinery of Scots pine (Pinus sylvestris) trees by monitoring multi-year changes in weather, chlorophyll fluorescence, chloroplast ultrastructure, and changes in pigment-protein composition. Analysis of Photosystem II and Photosystem I performance parameters indicate that highly dynamic structural and functional seasonal rearrangements of the photosynthetic apparatus occur. Although several mechanisms might contribute to 'sustained quenching' of winter/early spring pine needles, time-resolved fluorescence analysis shows that extreme down-regulation of photosystem II activity along with direct energy transfer from photosystem II to photosystem I play a major role. This mechanism is enabled by extensive thylakoid destacking allowing for the mixing of PSII with PSI complexes. These two linked phenomena play crucial roles in winter acclimation and protection.

Drought alters carbon (C) allocation within trees, thereby impairing tree growth. Recovery of root and leaf functioning and prioritized C supply to sink tissues after drought may compensate for drought-induced reduction of assimilation and growth. It remains unclear if C allocation to sink tissues during and following drought is controlled by altered sink metabolic activities or by the availability of new assimilates. Understanding such mechanisms is required to predict forests' resilience to a changing climate. We investigated the impact of drought and drought release on C allocation in a 100-y-old Scots pine forest. We applied 13 CO 2 pulse labeling to naturally dry control and long-term irrigated trees and tracked the fate of the label in above- and belowground C pools and fluxes. Allocation of new assimilates belowground was ca. 53% lower under nonirrigated conditions. A short rainfall event, which led to a temporary increase in the soil water content (SWC) in the topsoil, strongly increased the amounts of C transported belowground in the nonirrigated plots to values comparable to those in the irrigated plots. This switch in allocation patterns was congruent with a tipping point at around 15% SWC in the response of the respiratory activity of soil microbes. These results indicate that the metabolic sink activity in the rhizosphere and its modulation by soil moisture can drive C allocation within adult trees and ecosystems. Even a subtle increase in soil moisture can lead to a rapid recovery of belowground functions that in turn affects the direction of C transport in trees., Competing Interests: The authors declare no competing interest.

Exposure to lead (Pb) can have serious toxic effects on the physiological and biochemical processes of plants. The chemical form of the metal determines the degree of its toxicity. In our research, we examined the effect of lead in the form of lead nitrate [Pb(NO3)2] and lead chloride (PbCl2) in concentrations of 12.5 mM and 25 mM on pine (Pinus sylvestris) seed germination. Nitrogen salt causes more severe changes than chloride salt. Increasing levels of electrolyte leakage, malondialdehyde, and hydrogen peroxide were detected during germination processes. The high levels of ROS lead to redox changes in the cell. We observed a reduction in the level of the reduced form of glutathione (GSH), and at the same time observed increased levels of the oxidised form of glutathione (GSSG) depending on the concentration and also the time of exposure to lead compounds. At the beginning of germination processes, the effective non-enzymatic activity of the antioxidant cycle was dominant, and at the late stage the enzymatic activity was noticed in the presence of Pb compounds. CAT activity significantly increased after Pb compound exposition., Competing Interests: The authors have declared that no competing interests exist.

Tree stems are an overlooked source of volatile organic compounds (VOCs). Their contribution to ecosystem processes and total VOC fluxes is not well studied, and assessing it requires better understanding of stem emission dynamics and their driving processes. To gain more mechanistic insight into stem emission patterns, we measured monoterpene, methanol and acetaldehyde emissions from the stems of mature Scots pines (Pinus sylvestris L.) in a boreal forest over three summers. We analysed the effects of temperature, soil water content, tree water status, transpiration and growth on the VOC emissions and used generalized linear models to test their relative importance in explaining the emissions. We show that Scots pine stems are considerable sources of monoterpenes, methanol and acetaldehyde, and their emissions are strongly regulated by temperature. However, even small changes in water availability affected the emission potentials: increased soil water content increased the monoterpene emissions within a day, whereas acetaldehyde and methanol emissions responded within 2-4 days. This lag corresponded to their transport time in the xylem sap from the roots to the stem. Moreover, the emissions of monoterpenes, methanol and acetaldehyde were influenced by the cambial growth rate of the stem with 6-10-day lags., (© 2020 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Boreal forests are dominated by evergreen conifers that show strongly regulated seasonal photosynthetic activity. Understanding the mechanisms behind seasonal modulation of photosynthesis is crucial for predicting how these forests will respond to changes in seasonal patterns and how this will affect their role in the terrestrial carbon cycle. We demonstrate that the two co-occurring dominant boreal conifers, Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies), use contrasting mechanisms to reactivate photosynthesis in the spring. Scots pine downregulates its capacity for CO 2 assimilation during winter and activates alternative electron sinks through accumulation of PGR5 and PGRL1 during early spring until the capacity for CO 2 assimilation is recovered. In contrast, Norway spruce lacks this ability to actively switch between different electron sinks over the year and as a consequence suffers severe photooxidative damage during the critical spring period.

Stressors of different natures, including drought stress, substantially compromise the ability of plants to effectively and safely utilize light energy. We investigated the influence of water stress on the photosynthetic processes in Picea abies and Pinus sylvestris, two species with contrasting drought sensitivities. Spruce and pine seedlings were exposed to polyethylene glycol 6000-induced water deficits of different intensities and durations. The maintenance of photosystem I (PSI) oxidation in spruce required increased photosynthetic control and led to the increased reduction of the plastoquinone pool, which was not the case in pine seedlings. As a result of increased excitation pressure, photosystem II (PSII) inactivation was observed in spruce plants, whereas in pine, the decreased PSII photochemistry was likely due to sustained non-photochemical quenching. Downregulation of PSII photochemistry and maintenance of PSI in an oxidized state were linked with the prevention of oxidative stress, even under severe water deficit. The decreased photosynthetic pigment content and photosynthetic gene expression suggested the coordinated downregulation of photosynthetic apparatus components under water stress to reduce light energy absorption. In summary, the observed adaptative mechanisms of pine and spruce to water stress may be similar to the well-studied adaptative mechanisms to winter stress, which may indicate the universality of protective mechanisms under various stresses in conifers., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Decomposition is a major flux of the carbon cycle in forest soils and understanding the involved processes is a key for budgeting carbon turnover. Decomposition is constrained by the presence of biological agents such as microorganisms and the underlying environmental conditions such as water availability. A metabarcoding approach of ribosomal markers was chosen to study the succession of bacterial and fungal decomposers on root litter. Litterbags containing pine roots were buried in a pine forest for two years and sequentially sampled. Decomposition and the associated communities were surveyed under ambient dry and long-term irrigation conditions. Early decomposition stages were characterized by the presence of fast-cycling microorganisms such as Bacteroidetes and Helotiales, which were then replaced by more specialized bacteria and litter-associated or parasitic groups such as Acidobacteria, white rots, and Pleosporales. This succession was likely driven by a decrease of easily degradable carbohydrates and a relative increase in persistent compounds such as lignin. We hypothesize that functional redundancy among the resident microbial taxa caused similar root decomposition rates in control and irrigated forest soils. These findings have important implications for drought-prone Alpine forests as frequent drought events reduce litter fall, but not litter decomposition, potentially resulting in lower carbon stocks.

Radioactive contamination of the natural areas is one of the most long-lasting anthropogenic impacts on the environment. Scots pine (Pinus sylvestris L.) is a promising organism for radiation-related research because of its high radiosensitivity, but the genome size of Pinacea species has imposed obstacles for high-throughput studies so far. In this work, we conducted the analysis of the de novo assembled transcriptome of Scots pine populations growing in the Chernobyl-affected zone, which is still today contaminated with radionuclides because of the accident at the nuclear power plant in 1986. The transcriptome profiles indicate a clear pattern of adaptive stress response, which seems to be dose-dependent. The transcriptional response indicates a continuous modulation of the cellular redox system, enhanced expression of chaperones and histones, along with the control of ions balance. Interestingly, the activity of transposable element families is inversely correlated to the exposure levels to radiation. These adaptive responses, which are triggered by radiation doses 30 times lower than the one accepted as a safe for biota species by international regulations, suggest that the environmental management in radiation protection should be reviewed., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Abscisic acid (ABA) is one of the main participants in the regulation of plant responses to water deficiency. Knowledge of the ABA signal transduction pathways in gymnosperms is rather limited, especially in comparison with those in angiosperms. Seedlings of Scots pine and Norway spruce are known for their contrasting behaviour strategies under water deficit. To characterize the possible role of ABA in these differences, ABA dynamics were investigated under conditions of water deficit in seedlings of these two species. The content of ABA and its catabolites was followed in the roots and needles of seedlings of Pinus sylvestris and Picea abies under conditions of polyethylene glycol (PEG)-induced water deficiency (-0.15 and -0.5 MPa) for 10 days. The expression of the main genes for ABA-biosynthetic enzymes was also analysed. ABA showed more pronounced stress-dependent dynamics in pine roots than in spruce roots, whereas in needles, the response was greater for spruce than pine. The ABA increase during drought was mainly due to de novo synthesis and the shift in the balance between ABA synthesis and catabolism towards synthesis. The ABA-glucosyl ester did not serve as a reserve for the release of free ABA under water deficiency. The expression levels of the main ABA biosynthetic genes showed a weak or no correlation with changes in ABA content under water stress, i.e., the ABA content in the seedlings of both species was not directly linked to the transcript levels of the main ABA biosynthetic genes. Less-pronounced stress-induced changes in ABA in pine needles than in spruce needles may be related to pine seedlings having a less conservative strategy of growth and maintenance of water balance under water deficit., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Many studies have addressed several plant-insect interaction topics at nutritional, molecular, physiological, and evolutionary levels. However, it is still unknown how flexible the metabolism and the nutritional content of specialist insect herbivores feeding on different closely related plants can be. We performed elemental, stoichiometric, and metabolomics analyses on leaves of two coexisting Pinus sylvestris subspecies and on their main insect herbivore; the caterpillar of the processionary moth ( Thaumetopoea pityocampa ). Caterpillars feeding on different pine subspecies had distinct overall metabolome structure, accounting for over 10% of the total variability. Although plants and insects have very divergent metabolomes, caterpillars showed certain resemblance to their plant-host metabolome. In addition, few plant-related secondary metabolites were found accumulated in caterpillar tissues which could potentially be used for self-defense. Caterpillars feeding on N and P richer needles had lower N and P tissue concentration and higher C:N and C:P ratios, suggesting that nutrient transfer is not necessarily linear through trophic levels and other plant-metabolic factors could be interfering. This exploratory study showed that little chemical differences between plant food sources can impact the overall metabolome of specialist insect herbivores. Significant nutritional shifts in herbivore tissues could lead to larger changes of the trophic web structure.

Insectivorous birds feed upon all developmental stages of herbivorous insects, including insect eggs if larvae and adults are unavailable. Insect egg deposition on plants can induce plant traits that are subsequently exploited by egg parasitoids searching for hosts. However, it is unknown whether avian predators can also use egg-induced plant changes for prey localization. Here, we studied whether great tits (Parus major) and blue tits (Cyanistes caeruleus) are attracted by traits of the Scots pine (Pinus sylvestris) induced by pine sawfly (Diprion pini) egg deposition. We chose this plant - insect system because sawfly egg deposition on pine needles is known to locally and systemically induce a change in pine volatile organic compounds (VOCs), and tits are known to prey upon sawfly eggs. In dual choice laboratory experiments, we simultaneously offered the birds an egg-free control branch and a systemically egg-induced branch. Significantly more birds visited the egg-induced branch first. We confirmed by GC-MS analyses that systemically egg-induced branches released more (E)-β-farnesene compared to control branches. Spectrophotometric analyses showed that control branches reflected more light than egg-induced branches throughout the avian visual range. Although a discrimination threshold model for blue tits suggests that the birds are poor at discriminating this visual difference, the role of visual stimuli in attracting the birds to egg-induced pines cannot be discounted. Our study shows, for the first time, that egg-induced odorous and/or visual plant traits can help birds to locate insect eggs without smelling or seeing those eggs.

Long-lived conifers are vulnerable to climate change because classical evolutionary processes are slow in developing adaptive responses. Therefore, the capacity of a genotype to adopt different phenotypes is important. Gene expression is the primary mechanism that converts genome-encoded information into phenotypes, and DNA methylation is employed in the epigenetic regulation of gene expression. We investigated variations in global DNA methylation and gene expression between three Scots pine (Pinus sylvestris L.) populations located in northern and southern Finland using mature seeds. Gene expression levels were studied in six DNA methyltransferase (DNMT) genes, which were characterized in this study, and in 19 circadian clock genes regulating adaptive traits. In embryos, expression diversity was found for three DNMT genes, which maintain DNA methylation. The expression of two DNMT genes was strongly correlated with climate variables, which suggests a role for DNA methylation in local adaptation. For adaptation-related genes, expression levels showed between-population variation in 11 genes in megagametophytes and in eight genes in embryos, and many of these genes were linked to climate factors. Altogether, our results suggest that differential DNA methylation and gene expression contribute to local adaptation in Scots pine populations and may enhance the fitness of trees under rapidly changing climatic conditions.

Most plant-based emissions of volatile organic compounds are considered mainly temperature dependent. However, certain oxygenated volatile organic compounds (OVOCs) have high water solubility; thus, also stomatal conductance could regulate their emissions from shoots. Due to their water solubility and sources in stem and roots, it has also been suggested that their emissions could be affected by transport in the xylem sap. Yet further understanding on the role of transport has been lacking until present. We used shoot-scale long-term dynamic flux data from Scots pines (Pinus sylvestris) to analyse the effects of transpiration and transport in xylem sap flow on emissions of 3 water-soluble OVOCs: methanol, acetone, and acetaldehyde. We found a direct effect of transpiration on the shoot emissions of the 3 OVOCs. The emissions were best explained by a regression model that combined linear transpiration and exponential temperature effects. In addition, a structural equation model indicated that stomatal conductance affects emissions mainly indirectly, by regulating transpiration. A part of the temperature's effect is also indirect. The tight coupling of shoot emissions to transpiration clearly evidences that these OVOCs are transported in the xylem sap from their sources in roots and stem to leaves and to ambient air., (© 2018 John Wiley & Sons Ltd.)

The transition from the living water-transporting sapwood to heartwood involves in many tree species impregnation with extractives. These differ in amount and composition, and enhance resistance against bacteria, insects or fungi. To understand the synthesis, transport and impregnation processes new insights into the biochemical processes are needed by in-situ methods. Here we show the extractive distribution in pine (Pinus sylvestris) microsections with a high lateral resolution sampled in a non-destructive manner using Confocal Raman Microscopy. Integrating marker bands of stilbenes and lipids enables to clearly track the rapid change from sapwood to heartwood within one tree ring. The higher impregnation of the cell corner, compound middle lamella, the S3 layer and pits reveals the optimization of decay resistance on the micron-level. Furthermore, deposits with changing chemical composition are elucidated in the rays and lumen of the tracheids. The spectral signature of these deposits shows the co-location of lipids and pinosylvins with changing ratios from the living to the dead tissue. The results demonstrate that the extractive impregnation on the micro- and nano-level is optimized by a symbiotic relationship of lipids and pinosylvins to enhance the tree's resistance and lifetime.

Nonstructural carbohydrates (NSCs) are important for the growth and survival of trees. Drought may lead to a decrease in tree growth and to NSC depletion, whereas increased soil moisture in otherwise dry ecosystems may increase growth and NSC concentrations. A long-term (13 yr) irrigation experiment was conducted in a Pinus sylvestris-dominated forest located at the dry margin of the species in southern Switzerland. We measured the relative leaf area, growth, NSCs, needle δ 13 C, [N] and [P] in trees on control and irrigated plots. Irrigation resulted in higher growth rates and carbon isotope discrimination, but did not alter NSC levels. Growth and NSC decreased with decreasing leaf area in both treatments, but NSC did not correlate with leaf-level gas exchange indices, such as foliar δ 13 C, [N] or [P]. A legacy effect was shown, as trees with initially low leaf area had limited ability to respond to prolonged irrigation. The NSC constancy across treatments provides evidence that carbohydrate storage may stay constant when climate changes are sufficiently slow to allow acclimation. Moreover, we speculate that total leaf area, rather than leaf gas exchange per unit leaf area, drives the variation in whole-tree carbohydrate dynamics in this system., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

Arctic moistening will affect the circumpolar forested riparian ecosystems. Upward trends observed for precipitation in high latitudes illustrate that the moistening may be underway to influence the woody biomass production near the inland waters, lakes and streams with effects on carbon pools and fluxes. Although the flooding and waterlogging tolerance of seedlings has been investigated, our understanding of responses in mature trees is still limited. Here we employ tree-ring δ 13 C and width data from a subarctic riparian setting in Lapland, where artificially high lake level (HLL) has already altered the ecophysiological and growth responses of riparian Pinus sylvestris trees to external drivers under conditions simulating moister environment. Prior to the HLL event, the carbon assimilation rate was primarily limited by irradiance as reflected in the δ 13 C data and the radial growth of south-facing riparian trees remained increased in comparison to shaded upland trees. By contrast, the riparian trees were not similarly benefited during the HLL period when reduced assimilation depleted the riparian in comparison to upland δ 13 C despite of increased irradiance. As a result, the radial growth of riparian trees was markedly reduced over the HLL event while the upland trees benefited from increased irradiance and summer time warming. Although the production of biomass at high latitudes is commonly considered temperature-limited, our results highlight the increasing role of Arctic moistening to limit the growth when increased precipitation (cloudiness) reduces the incoming solar radiation in general and when the riparian habitat becomes increasingly waterlogged in particular. The effects of high-latitude warming to induce higher biomass productivity may be restricted by negative feedbacks., (© 2018 John Wiley & Sons Ltd.)

Background and Aims: Polyamines are small metabolites present in all living cells and play fundamental roles in numerous physiological events in plants. The aminopropyltransferases (APTs), spermidine synthase (SPDS), spermine synthase (SPMS) and thermospermine synthase (ACL5), are essential enzymes in the polyamine biosynthesis pathway. In angiosperms, SPMS has evolved from SPDS via gene duplication, whereas in gymnosperms APTs are mostly unexplored and no SPMS gene has been reported. The present study aimed to investigate the functional properties of the SPDS and ACL5 proteins of Scots pine (Pinus sylvestris L.) in order to elucidate the role and evolution of APTs in higher plants., Methods: Germinating Scots pine seeds and seedlings were analysed for polyamines by high-performance liquid chromatography (HPLC) and the expression of PsSPDS and PsACL5 genes by in situ hybridization. Recombinant proteins of PsSPDS and PsACL5 were produced and investigated for functional properties. Also gene structures, promoter regions and phylogenetic relationships of PsSPDS and PsACL5 genes were analysed., Key Results: Scots pine tissues were found to contain spermidine, spermine and thermospermine. PsSPDS enzyme catalysed synthesis of both spermidine and spermine. PsACL5 was found to produce thermospermine, and PsACL5 gene expression was localized in the developing procambium in embryos and tracheary elements in seedlings., Conclusions: Contrary to previous views, our results demonstrate that SPMS activity is not a novel feature developed solely in the angiosperm lineage of seed plants but also exists as a secondary property in the Scots pine SPDS enzyme. The discovery of bifunctional SPDS from an evolutionarily old conifer reveals the missing link in the evolution of the polyamine biosynthesis pathway. The finding emphasizes the importance of pre-existing secondary functions in the evolution of new enzyme activities via gene duplication. Our results also associate PsACL5 with the development of vascular structures in Scots pine.

Phytopathogenic fungi infections induce plant defence responses that mediate changes in metabolic and signalling processes with severe consequences for plant growth and development. Sphaeropsis tip blight, induced by the endophytic fungus Sphaeropsis sapinea that spreads from stem tissues to the needles, is the most widespread disease of conifer forests causing dramatic economic losses. However, metabolic consequences of this disease on bark and wood tissues of its host are largely unexplored. Here, we show that diseased host pines experience tissue dehydration in both bark and wood. Increased cytokinin and declined indole-3-acetic acid levels were observed in both tissues and increased jasmonic acid and abscisic acid levels exclusively in the wood. Increased lignin contents at the expense of holo-cellulose with declined structural biomass of the wood reflect cell wall fortification by S. sapinea infection. These changes are consistent with H 2 O 2 accumulation in the wood, required for lignin polymerization. Accumulation of H 2 O 2 was associated with more oxidized redox states of glutathione and ascorbate pools. These findings indicate that S. sapinea affects both phytohormone signalling and the antioxidative defence system in stem tissues of its pine host during the infection process., (© 2017 John Wiley & Sons Ltd.)

A scanning electron microscopy (SEM) investigation of pine (Pinus sylvestris) and oak (Quercus sp.) wood samples exposed to various types of natural degradation is presented with the aim of discussing the correct identification of multiple degradation signs in waterlogged wood. This is part of an experiment performed at the archeological site of Biskupin (Poland) to evaluate the dynamics of short-term wood degradation during reburial and the suitability of excavated wood as substrate for the fungal attack. The final aim is to support and inform the in situ conservation strategy currently applied to archeological woods. To replicate the burial conditions, wood samples were put into lake water and peat. The samples were removed from the burial environments after 4, 6, 8, and 10 years, and then exposed to laboratory-controlled attack by a brown rot fungus Coniophora puteana and a white rot fungus Coriolus versicolor. SEM images were acquired for all samples before and after the fungal attack. The results showed a slight degradation occurred in the burial environments (soft rot and bacteria). In addition, both typical and previously neglected features of fungal attack were observed, highlighting that the extent of the fungal decay varies according to the previous degree of wood degradation. Some comparisons are provided with archeological wood samples from the Biskupin site.

A key weakness in current Earth System Models is the representation of thermal acclimation of photosynthesis in response to changes in growth temperatures. Previous studies in boreal and temperate ecosystems have shown leaf-scale photosynthetic capacity parameters, the maximum rates of carboxylation (V cmax ) and electron transport (J max ), to be positively correlated with foliar nitrogen (N) content at a given reference temperature. It is also known that V cmax and J max exhibit temperature optima that are affected by various environmental factors and, further, that N partitioning among the foliar photosynthetic pools is affected by N availability. However, despite the strong recent anthropogenic influence on atmospheric temperatures and N deposition to forests, little is known about the role of foliar N contents in controlling the photosynthetic temperature responses. In this study, we investigated the temperature dependencies of V cmax and J max in 1-year-old needles of mature boreal Pinus sylvestris (Scots pine) trees growing under low and high N availabilities in northern Sweden. We found that needle N status did not significantly affect the temperature responses of V cmax or J max when the responses were fitted to a peaked function. If such N insensitivity is a common tree trait it will simplify the interpretation of the results from gradient and multi-species studies, which commonly use sites with differing N availabilities, on temperature acclimation of photosynthetic capacity. Moreover, it will simplify modeling efforts aimed at understanding future carbon uptake by precluding the need to adjust the shape of the temperature response curves to variation in N availability., (© 2017 Scandinavian Plant Physiology Society.)

We investigated the transfer of 15 N into the soil via 15 N uptake and release by tree roots, which involves the principles of the split-root technique. One half of the root system received an injection of ( 15 NH 4 ) 2 SO 4 and the other half equivalent amounts of (NH 4 ) 2 SO 4 at 15 N natural abundance level. 15 N was transferred from one side of the root system ( 15 N side) to the other side ( 14 N side) and released into the soil. The method was conducted with Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies [L.] Karst). Two concentration levels of (NH 4 ) 2 SO 4 were used, corresponding with annual N deposition in the Netherlands (30 kg N ha -1 ) and a twelfth of that (2.5 kg N ha -1 ). Samples were taken 3 and 6 weeks after labelling and divided into needles + stem, roots, rhizosphere and bulk soil. Already 3 weeks after labelling, Scots pine took up 23.7 % of the low and 9.1 % of the high amounts of 15 N, while Norway spruce took up 21.5 and 32.1 %, respectively. Both species transported proportions of 15 N to the rhizosphere (0.1-0.2 %) and bulk soil (0.3-0.9 %). The method is a useful tool to investigate the fate of root-derived N in soils, for example, for the formation of stable forms of soil organic matter.

Secondary organic aerosol (SOA) impact climate by scattering and absorbing radiation and contributing to cloud formation. SOA models are based on studies of simplified chemical systems that do not account for the chemical complexity in the atmosphere. This study investigated SOA formation from a mixture of real Scots pine (Pinus sylvestris) emissions including a variety of monoterpenes and sesquiterpenes. SOA generation was characterized from different combinations of volatile compounds as the plant emissions were altered with an herbivore stress treatment. During active herbivore feeding, monoterpene and sesquiterpene emissions increased, but SOA mass yields decreased after accounting for absorption effects. SOA mass yields were controlled by sesquiterpene emissions in healthy plants. In contrast, SOA mass yields from stressed plant emissions were controlled by the specific blend of monoterpene emissions. Conservative estimates using a box model approach showed a 1.5- to 2.3-fold aerosol enhancement when the terpene complexity was taken into account. This enhancement was relative to the commonly used model monoterpene, "α-pinene". These results suggest that simplifying terpene complexity in SOA models could lead to underpredictions in aerosol mass loading.

Trees of Scots pine (Pinus sylvestris L.) are known for their effective phytoextraction capabilities. The results obtained in this study point to the significant role of substrate composition and chemical characteristics in the phytoextraction potential of this species. A multi-elemental (53 elements) analysis of pines from unpolluted (soil) and polluted (post-flotation tailings) sites was performed using inductively coupled plasma optical emission spectrometry. The analyzed flotation tailings were characterized by alkaline pH (7.19 ± 0.06) and significantly higher conductivity (277.7 ± 2.9 µS cm -1 ) than the soil (pH = 5.11 ± 0.09; 81.3 ± 4.9 µS cm -1 ). The two substrates also differed with respect to the contribution of the clay fraction (0% in the unpolluted and 8% in the polluted substrate). The specimens of P. sylvestris growing on flotation tailings had significantly smaller height (381 ± 58 cm) and total aboveground biomass (4.78 ± 0.66 kg) than the trees growing in soil (699 ± 80 cm and 10.24 ± 2.10 kg). The biomass of the trunk, twigs and branches, and needles of the trees from polluted sites was between 40.0% and 48.7% of the biomass of the same organs of the control trees. Generally, the organs (trunk, twigs and branches, needles) of the P. sylvestris specimens from polluted sites had significantly higher concentrations of Au, Al, Ba, Cd, Co, La, Lu, Ni, Pd, Sc, Zn, and lower concentrations of B, Bi, Ca, Ce, Er, In, K, Mg, Na, Nd, P, Pr, Re, Se, Sr, Te than in the control plants, these metals being accumulated effectively in the whole of the aboveground biomass (BCF>1). Although the concentration of the majority of elements was significantly higher in the flotation tailings, significantly higher concentrations of these elements were observed in the tree organs from unpolluted sites, which points to the important role of substrate characteristics in the phytoextraction efficiency of P. sylvestris.

We report results from long-term simulated external nitrogen (N) input experiments in three northern Pinus sylvestris forests, two of moderately high and one of moderately low productivity, assessing effects on annual net primary production (NPP) of woody mass and its interannual variation in response to variability in weather conditions. A sigmoidal response of wood NPP to external N inputs was observed in the both higher and lower productivity stands, reaching a maximum of ~65% enhancement regardless of the native site productivity, saturating at an external N input of 4-5 g N·m -2 ·yr -1 . The rate of increase in wood NPP and the N response efficiency (RE N , increase in wood NPP per external N input) were maximized at an external N input of ~3 g N·m -2 ·yr -1 , regardless of site productivity. The maximum RE N was greater in the higher productivity than the lower productivity stand (~20 vs. ~14 g C/g N). The N-induced enhancement of wood NPP and its RE N were, however, markedly contingent on climatic variables. In both of the higher and lower productivity stands, wood NPP increased with growing season precipitation (P), but only up to ~400 mm. The sensitivity of the response to P increased with increasing external N inputs. Increasing growing season temperature (T) somewhat increased the N-induced drought effect, whereas decreasing T reduced the drought effect. These responses of wood NPP infused a large temporal variation to RE N , making the use of a fixed value unadvisable. Based on these results, we suggest that regional climate conditions and future climate scenarios should be considered when modeling carbon sequestration in response to N deposition in boreal P. sylvestris, and possibly other forests., (© 2017 by the Ecological Society of America.)

To predict how the function of urban vegetation and the provision of ecosystem services respond to combinations of natural and anthropogenic drivers, a better understanding of multiple stress interactions is required. This study tested combined effects of moderate levels of drought, soil salinity and exposure to diesel exhaust on parameters of physiology, metabolism, morphology and growth of Pinus sylvestris L. saplings. We found that plant responses were primarily dominated by single stressors and a few two-way interactions. Stressor combinations did not have considerable additional negative effects on plant performance compared to single stressors. Hence, synergistic and antagonistic interactions were rare and additive effects frequent. Drought cycles caused most negative effects, from chlorophyll a fluorescence and epicuticular wax content to growth responses, while soil salinity caused fewer negative effects but contributed to reduction in fine root growth and fluorescence parameters at low air contamination. Interestingly, the air contamination alone had only marginal effects on plant morphology and growth, but contributed an antagonistic effect, dampening the negative effect of drought and salinity on the maximum quantum efficiency of PSII photochemistry (F v /F m ) and fine root biomass. Although, these effects were moderate, it appears that exhaust exposure had a cross-acclimation effect on plant responses to drought and salinity. We also found that salinity had a negative effect on the accumulation of particulate matter on shoots, illustrating that the plant stress situation can affect the provisioning of certain ecosystem services like pollution attenuation. These findings have implications for the understanding of the complex natural and anthropogenic stress situation of urban, and how to maintain the ecological functions and delivery of ecosystem services., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Carbon reserves are important for maintaining tree function during and after stress. Increasing tree mortality driven by drought globally has renewed the interest in how plants regulate allocation of recently fixed C to reserve formation. Three-year-old seedlings of two species (Tilia platyphyllos and Pinus sylvestris) were exposed to two intensities of experimental drought during ~10 weeks, and 13 C pulse labelling was subsequently applied with rewetting. Tracking the 13 C label across different organs and C compounds (soluble sugars, starch, myo-inositol, lipids and cellulose), together with the monitoring of gas exchange and C mass balances over time, allowed for the identification of variations in C allocation priorities and tree C balances that are associated with drought effects and subsequent drought release. The results demonstrate that soluble sugars accumulated in P. sylvestris under drought conditions independently of growth trends; thus, non-structural carbohydrates (NSC) formation cannot be simply considered a passive overflow process in this species. Once drought ceased, C allocation to storage was still prioritized at the expense of growth, which suggested the presence of 'drought memory effects', possibly to ensure future growth and survival. On the contrary, NSC and growth dynamics in T. platyphyllos were consistent with a passive (overflow) view of NSC formation., (© 2017 John Wiley & Sons Ltd.)

Plants have developed a complex defense response system against pests and pathogens. Defensins, produced by plants as part of their innate immune response, form the family of small, basic, cysteine-rich proteins with activity primarily directed against fungal pathogens. In addition, plant defensins can show antibacterial activity and protease and insect amylase inhibitory activities. However, in gymnosperms, only antifungal activity of defensins has been described thus far. Here, we report antibacterial and insect α-amylase inhibition activities for defensin PsDef1 from P. sylvestris, the first defensin from gymnosperms with a broad range of biological activities described. We also report the solution NMR structure of PsDef1 and its dynamics properties assessed by a combination of experimental NMR and computational techniques. Collectively, our data provide an insight into structure, dynamics, and functional properties of PsDef1 that could be common between defensins from this taxonomic group., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Heartwood extractives are important determinants of the natural durability of pine heartwood. The most important phenolic compounds affecting durability are the stilbenes pinosylvin and its monomethylether, which in addition have important functions as phytoalexins in active defense. A substantial portion of the synthesized pinosylvin is 3-methoxylated but the O-methyltransferase responsible for this modification has not been correctly identified. We studied the expression of the stilbene pathway during heartwood development as well as in response to wounding of xylem and UV-C treatment of needles. We isolated and enzymatically characterized a novel O-methyltransferase, PMT2. The methylated product was verified as pinosylvin monomethylether using ultra performance liquid chromatography-tandem mass spectrometry and high performance liquid chromatography analyses. The PMT2 enzyme was highly specific for stilbenes as substrate, in contrast to caffeoyl-CoA O-methyltransferase (CCoAOMT) and PMT1 that were multifunctional. Expression profile and multifunctional activity of CCoAOMT suggest that it might have additional roles outside lignin biosynthesis. PMT1 is not involved in the stilbene pathway and its biological function remains an open question. We isolated a new specific O-methyltransferase responsible for 3-methoxylation of pinosylvin. Expression of PMT2 closely follows stilbene biosynthesis during developmental and stress induction. We propose that PMT2 is responsible for pinosylvin methylation in Scots pine (Pinus sylvestris), instead of the previously characterized methyltransferase, PMT1., (© 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.)

This is the first study to investigate stress proteins dehydrins with the use of specific antibodies in the Scots pine (Pinus sylvestris L.) needles and their changes in the annual cycle under extreme climate of Yakutia. No pronounced polymorphism of major dehydrins (14-15 and 66 kDa) has been found during the winter dormancy period of P. sylvestris. A clear correlation between the seasonal variations in dehydrins and changes in the water content in needles was revealed. Consistently high levels of dehydrins was retained throughout the period of low negative temperatures. It is assumed that dehydrins can participate in the formation of P. sylvestris L. resistance to the permafrost conditions.

Plants have an accumulative response to heavy metals present in soils or deposited from airborne sources of emissions. Therefore, their tissues are very often used in studies of heavy metal contamination originating from different sources as a bioindicator of environmental pollution. This research was undertaken to examine accumulation capacities of Pb, Zn, Cd, Cu and Cr in washed and unwashed needles of Scots pine (Pinus sylvestris L.) and leaves of silver birch (Betula pendula Roth) growing in a contaminated area. We collected needles of Scots pine and leaves of silver birch in an area around a sedimentation pond and metallurgic plant processing Pb and Zn ores near Olkusz, Poland. Concentrations of heavy metals, which have been linked with exposure to emissions, were determined from foliar samples collected at 33 sites. These sites were established at various distances (0.5-3.6 km) from the pond and metallurgic plant so as to identify the predominant accumulative response of plants. Spatial gradients for Pb and Zn were calculated using an ordinary kriging interpolation algorithm. A spatial pattern was identified by a GIS method to visualize maps over the Pb-Zn ore mining area. The accumulation of Zn (R 2  = 0.74, p < 0.05) and Pb (R 2  = 0.85, p < 0.01) in plant tissues correlated with soil concentrations. This tendency was not found in the case of Cu, Cd and Cr., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Scots pine (Pinus sylvestris L.) provenances cover broad ecological amplitudes. In a greenhouse study, we investigated the impact of drought stress and rewetting on gas exchange for three provenances (Italy: Emilia Romagna; Spain: Alto Ebro; Germany: East-German lowlands) of 2-year old Scots pine seedlings. CO 2 , water vapour and isoprenoid exchange of stressed and control trees were quantified with a four-chamber dynamic-enclosure system in the controlled environment of a climate chamber. The three provenances showed distinct isoprenoid emission patterns and were classified into a non-Δ 3 -carene, with either high α-/β-pinene or β-myrcene fraction, and a Δ 3 -carene dominated type. Isoprenoid emission rates, net-photosynthesis and transpiration were reduced during summer drought stress and significantly recovered after rewetting. A seasonal increase of isoprenoid emission rates towards autumn was observed for all control groups. Compared with the German provenance, the Spanish and Italian provenances revealed higher isoprenoid emission rates and more plastic responses to drought stress and seasonal development, which points to a local adaptation to climate. As a result of drought, net carbon uptake and transpiration of trees was reduced, but recovered after rewetting. We conclude from our study that Scots pine isoprenoid emission is more variable than expected and sensitive to drought periods, likely impacting regional air chemistry. Thus, a provenance-specific emission assessment accounting for reduced emission during prolonged (summer) drought is recommend for setting up biogenic volatile organic compound emission inventories used in air quality models., (© The Author 2016. Published by Oxford University Press.)