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Colonies of piscivorous birds nesting in forests bring in loads of allochthonous material in amounts significantly exceeding atmospheric depositions, eventually causing major ecosystem changes. We studied the effect of increasing ornithogenic impact on ectomycorrhizal (EMF) and other root-associated fungi in a Scots pine forest affected by a colony of great cormorants. We evaluated quantitative data of ectomycorrhiza (numbers of root tips and morphotypes) and identified fungal species from pine root tips and from sporocarps collected in three designated study zones: D (zone of active nesting), E (colony margin), and G (pristine forest). Species-bearing sporocarps were absent in zone D, and their number steadily decreased in zone E and was significantly lower than in zone G. Species communities and occurrence frequencies of individual species differed significantly between the zones. Environmental factors, both directly (nutrient addition) and indirectly (changes in vegetation cover), associated with bird activity have significantly influenced fungal communities. We hypothesize that the first signs of increased avian impact on root-associated fungi are the reduced diversity of sporocarps and the low presence and diversity of boletoid fungi in root tips.

Pinus sylvestris is an important production tree. In recent years, there has been a sharp increase in the mortality of pine trees due to insect pests. It is obvious that some pests profit from climatic changes, increase their aggressiveness and spread to new localities. The study aimed to investigate the spectrum of more abundant insect pest species in pine plantations of Czechia. The occurrence of species and intensity of their infestation were studied at 77 localities situated in six regions. Any abundant foliophagous insect species were noticed. Bark beetles and wood-boring insects were found to be prevalent. Namely, Ips acuminatus, Ips sexdentatus, Phaenops cyanea and Sirex noctilio seem to be the most dangerous. These species are now better competitors than species previously considered as main pests. Their aggressiveness, expansion to new areas and interspecies co-occurrence are alarming. Due to the advancing climate change, pine mortality due to bark beetles and wood-boring pests will probably continue to increase.

Dehydrins are well-known components of plant responses to different stresses that cause dehydration, including drought, freezing, salinity, etc. In conifers, the dehydrin gene family is very large, implying that the members of this family have important physiological functions in conifer stress tolerance. However, dehydrin gene expression displays a wide range of responses to stress, from thousand-fold increased expression to decreased expression, and it is generally unknown how regulatory systems are connected at the mRNA and protein levels. Therefore, we studied these aspects of dehydrin regulation in Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) H. Karst) seedlings under polyethylene glycol 6000-induced osmotic stress ranging from relatively low (culture medium water potential of −0.15 MPa) to very high (−1.0 MPa) intensities. In pine, the major dehydrin protein was Dhn1 in both the roots and needles, and in spruce, two isoforms of the Dhn4 protein were the major dehydrins; both of these proteins are AESK-type dehydrins. The genes encoding these major proteins were highly expressed even under control conditions; surprisingly, we also observed several highly expressed dehydrin genes that were not abundantly translated. Under osmotic stress, the most prominent expression changes were observed for the dehydrin genes with low basal expression levels, whereas highly expressed genes generally demonstrated rather modest changes in expression. We report proposed constitutive physiological functions of the AESK-type dehydrins in Pinaceae plants.

Background: Site productivity remains a fundamental concern in forestry as a significant driver of resource availability. The site index (SI) reflects the overall impact of all environmental parameters that determine tree height growth and is the most commonly used indirect proxy for forest site productivity estimated using stand age and height. One of the most critical challenges in the site index (SI) concept are local variations in climate, soil, and genotype-environmental interactions that lead to variable height growth patterns among ecoregions and cause inappropriate estimation of site productivity. Developing regional models can solve this problem and allow us to determine forest growth and SI appropriately.Results: Therefore, the purpose of this study was to develop regional height growth models (RMs) for the Scots pine in Poland. For height growth modelling, we used the growth trajectory data of 855 sample trees, representing the entire range of geographic locations and site conditions of the Scots pine in Poland. Collected growth trajectories were used for the development of the global height growth model (GM) for Poland and RMs for six natural forest regions, which were adopted as the spatial unit for the model regionalisation. Height prediction errors by the global model were found to be significantly larger than those obtained with regional models in all regions. The results showed significant differences between growth trajectories in natural forest regions I, II, and III located in northern Poland compared to stands in natural forest regions IV, V, and VI in southern Poland.Conclusions: The presented study showed differences in height growth patterns of Scots pines in Poland and revealed that the use of local models could improve the growth prediction and quality of the SI estimation. Developed RMs show better fit statistics and predictive validity than the GM developed for the countrywide scale. Differences in climate and soil conditions which distinguish natural forest regions affect height growth patterns of Scots pine. Therefore, extending this research to models which directly describe the interactions of height growth with site variables, such as climate, soil properties, and topography, can provide additional valuable forest management information.

Trees have a strong influence on the chemical properties of the soil in which they grow. Establishing plantations with different tree species combinations thus potentially leads to divergence in soil chemistry. To study the degree to which differentiation already occurs during the first years after establishment, we made use of a biodiversity–ecosystem functioning experiment in Belgium, FORBIO. The multi-site experiment replicates tree species richness (1–4 species) and composition in three sites. The sites represent contrasting site contexts, mostly land-use history, soil, and climate. Soil samples (0–10 cm) were taken at the time of planting and approximately 8 years later. We measured the total C and N concentration, the Olsen P, the pH-H2O, and the concentration of base and Al cations. The change in chemical composition was strongly dependent on site conditions including former land use. Afforestation on former cropland had a positive impact on total C and Olsen P and a negative effect on base cations and pH. On sites reforested after clear-cut, soil texture and particular site preparation played an important role. On top of that, we found several significant effects of species composition. Strongest composition effects were detected in the reforested site on loamy soil with little soil disturbance. This study highlights that species choice can already affect soil chemistry in early stages of forest development, but that the nature of the effects may strongly depend on the context in which the plantations are established. Further research is needed to identify the most important contextual factors.

Interpreting phloem carbohydrate or xylem tissue carbon isotopic composition as measures of water-use efficiency or past tree productivity requires in-depth knowledge of the factors altering the isotopic composition within the pathway from ambient air to phloem contents and tree ring. One of least understood of these factors is mesophyll conductance (g(m)). We formulated a dynamic model describing the leaf photosynthetic pathway including seven alternative g(m) descriptions and a simple transport of sugars from foliage down the trunk. We parameterised the model for a boreal Scots pine stand and compared simulated g(m) responses with weather variations. We further compared the simulated delta C-13 of new photosynthates among the different g(m) descriptions and against measured phloem sugar delta C-13. Simulated g(m) estimates of the seven descriptions varied according to weather conditions, resulting in varying estimates of phloem delta C-13 during cold/moist and warm/dry periods. The model succeeded in predicting a drought response and a postdrought release in phloem sugar delta C-13 indicating suitability of the model for inverse prediction of leaf processes from phloem isotopic composition. We suggest short-interval phloem sampling during and after extreme weather conditions to distinguish between mesophyll conductance drivers for future model development.

Many plant species form symbioses with ectomycorrhizal fungi, which help them forage for limiting nutrients in the soil such as inorganic phosphate (Pi). The transcriptional responses to symbiosis and nutrient-limiting conditions in ectomycorrhizal fungal hyphae, however, are largely unknown. An artificial system was developed to study ectomycorrhizal basidiomycete Paxillus involutus growth in symbiosis with its host tree Pinus sylvestris at different Pi concentrations. RNA-seq analysis was performed on P. involutus hyphae growing under Pi-limiting conditions, either in symbiosis or alone. We show that Pi starvation and ectomycorrhizal symbiosis have an independent effect on the P. involutus transcriptome. Notably, low Pi availability induces expression of newly identified putative high-affinity Pi transporter genes, while reducing the expression of putative organic acid transporters. Additionally, low Pi availability induces a close transcriptional interplay between P and N metabolism. GTP-related signalling was found to have a positive effect in the maintenance of ectomycorrhizal symbiosis, whereas multiple putative cytochrome P450 genes were found to be downregulated, unlike arbuscular mycorrhizal fungi. We provide the first evidence of global transcriptional changes induced by low Pi availability and ectomycorrhizal symbiosis in the hyphae of P. involutus, revealing both similarities and differences with better-characterized arbuscular mycorrhizal fungi.

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 (CO2)-C-13 pulse labeling to naturally dry control and long-term irrigated trees and tracked the fate of the label in aboveand 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.

Pine sawflies (Hymenoptera: Diprionidae) belong to the most common pests of Scots pine Pinus sylvestris L. in the area of the Regional Directorates of State Forests in Toruń and Gdańsk. The two most destructive species, Diprion pini L. in the area of Puszcza Bydgoska and Gilpinia virens (Klug) in Bory Tucholskie, were investigated in this study. Our main aim was to characterise the two species in the Bydgoszcz Forest District in RDSF in Toruń and Kaliska in RDSF in Gdańsk, respectively, based on data gathered during outbreaks between 1991 and 2011. For this purpose, a geometric database was created, containing information about the year of outbreak, number of larvae observed after tree felling as well as selected stand and site features including stand age and species composition, stocking index, canopy closure, site index, humidity and fertility. By analysing the occurrence of both species in relation to stand and site features, we were able to determine their environmental requirements, which are quite different from one another. D. pini preferred older stands, with a stocking index between 0.8–0.9, growing on fresh and dry coniferous sites, while species composition (share of pine), canopy closure or site index did not have any influence on its abundance. G. virens preferred middle-aged pure pine stands, with relatively open canopies, on fresh coniferous sites and poorer soils. The stocking index did not affect its occurrence.

Understanding the consequences of local adaptation at the genomic diversity is a central goal in evolutionary genetics of natural populations. In species with large continuous geographical distributions the phenotypic signal of local adaptation is frequently clear, but the genetic background often remains elusive. We examined the patterns of genetic diversity inPinus sylvestris, a keystone species in many Eurasian ecosystems with a huge distribution range and decades of forestry research showing that it is locally adapted to the vast range of environmental conditions. MakingP. sylvestrisan even more attractive subject of local adaptation study, population structure has been shown to be weak previously and in this study. However, little is known about the molecular genetic basis of adaptation, as the massive size of gymnosperm genomes has prevented large scale genomic surveys. We generated a both geographically and genomically extensive dataset using a targeted sequencing approach. By applying divergence-based and landscape genomics methods we found that several coding loci contribute to local adaptation. We also discovered a very large (ca. 300 Mbp) putative inversion with a signal of local adaptation, which to our knowledge is the first such discovery in conifers. Our results call for more detailed analysis of structural variation in relation to genomic basis of local adaptation, emphasize the lack of large effect loci contributing to local adaptation in the coding regions and thus point out to the need for more attention towards multi-locus analysis of polygenic adaptation.

Climate warming enables tree seedling establishment beyond the current alpine treeline, but to achieve this, seedlings have to establish within existing tundra vegetation. In tundra, mosses are a prominent feature, known to regulate soil temperature and moisture through their physical structure and associated water retention capacity. Moss presence and species identity might therefore modify the impact of increases in temperature and precipitation on tree seedling establishment at the arctic‐alpine treeline. We followed Betula pubescens and Pinus sylvestris seedling survival and growth during three growing seasons in the field. Tree seedlings were transplanted along a natural precipitation gradient at the subarctic‐alpine treeline in northern Sweden, into plots dominated by each of three common moss species and exposed to combinations of moss removal and experimental warming by open‐top chambers (OTCs). Independent of climate, the presence of feather moss, but not Sphagnum , strongly supressed survival of both tree species. Positive effects of warming and precipitation on survival and growth of B. pubescens seedlings occurred in the absence of mosses and as expected, this was partly dependent on moss species. P. sylvestris survival was greatest at high precipitation, and this effect was more pronounced in Sphagnum than in feather moss plots irrespective of whether the mosses had been removed or not. Moss presence did not reduce the effects of OTCs on soil temperature. Mosses therefore modified seedling response to climate through other mechanisms, such as altered competition or nutrient availability. We conclude that both moss presence and species identity pose a strong control on seedling establishment at the alpine treeline, and that in some cases mosses weaken climate‐change effects on seedling establishment. Changes in moss abundance and species composition therefore have the potential to hamper treeline expansion induced by climate warming.

In many regions, drought is suspected to be a cause of Scots pine decline and mortality, but the underlying physiological mechanisms remain unclear. Because of their relationship to ecohydrological processes, δ18O values in tree rings are potentially useful for deciphering long-term physiological responses and tree adaptation to increasing drought. We therefore analyzed both needle- and stem-level isotope fractionations in mature trees exposed to varying water supply. In a first experiment, we investigated seasonal δ18O variations in soil and needle water of Scots pine in a dry inner Alpine valley in Switzerland, comparing drought-stressed trees with trees that were irrigated for more than 10 years. In a second experiment, we analyzed twentieth-century δ18O variations in tree rings of the same forest, including a group of trees that had recently died. We observed less 18O enrichment in needle water of drought-stressed compared with irrigated trees. We applied different isotope fractionation models to explain these results, including the Péclet and the two-pool correction, which considers the ratio of unenriched xylem water in the needles to total needle water. Based on anatomical measurements, we found this ratio to be unchanged in drought-stressed needles, although they were shorter. The observed lower 18O enrichment in needles of stressed trees was therefore likely caused by increased effective path length for water movement within the leaf lamina. In the tree-ring study, we observed lower δ18O values in tree rings of dead trees compared with survivors during several decades prior to their death. These lower values in declining trees are consistent with the lower needle water 18O enrichment observed for drought-stressed compared with irrigated trees, suggesting that this needle-level signal is reflected in the tree rings, although changes in rooting depth could also play a role. Our study demonstrates that long-term effects of drought are reflected in the tree-ring δ18O values, which helps to provide a better understanding of past tree physiological changes of Scots pine.

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 two to four 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 six- to ten-day lags. This article is protected by copyright. All rights reserved.

WOS:000540457600108 Chemical and thermal aspects of Pinus sylvestris and Pinus pinaster were characterized, with consideration of non-extracted and cylohexaneethanol (EtOH)-extracted woods, holocellulose, and alpha-cellulose. The chemical properties including cylohexane-EtOH solubility of wood were determined by standard test methods, holocellulose, alpha-cellulose, lignin, and ash components. Fourier transform infrared attenuated total reflection spectroscopic (FTIR-ATR) analysis revealed changes in the chemical structure of the specimens. Lignin peaks were still observed in holocelluloses' spectra, and substantial loss of polysaccharides were also detected. Moreover, the alpha-cellulose spectra of both softwoods included bands belonging to polyoses. Pinus pinaster's holocellulose, alpha-cellulose, and ash contents were higher than those of Pinus sylvestris but lignin and cylohexane-EtOH-solubility amounts were lower. The mass loss during degradation stage of Pinus sylvestris and Pinus pinaster was observed in the extracted wood respectively as 73.0 and 74.5% within the 188.7 to 422.9 degrees C range. The highest mass loss for Pinus sylvestris and Pinus pinaster in the carbonization step was formed in their holocelluloses respectively as 22.9 and 22.5% from 426 degrees C to 1017 degrees C.

Future climate scenarios for the boreal zone project increasing temperatures and precipitation, as well as extreme weather events such as heavy rain during the growing season. This can result in more frequent short-term waterlogging (WL) leading to unfavorable conditions for tree roots. In addition, it is decisive whether short-term WL periods during the growing season occur continuously or periodically. We assessed the effects of short-termed WL on 4-year-old Scots pine (Pinus sylvestris L.) saplings after shoot elongation started. Waterlogging (WL) lasted either continuously for 2.5 weeks (ContWL) or noncontinuously for 5 weeks, consisting of three repeated 1-week-interval WL periods (IntWL). Both treatments resulted in the same duration of soil anoxia. We studied soil gases, root and shoot growth and physiology, and root survival probability and longevity during the experiment. In the final harvest, we determined shoot and root biomass and hydraulic conductance and electrical impedance spectra of the root systems. Soil CO2 and CH4 concentrations increased immediately after WL onset and O2 decreased until anoxia. Waterlogging decreased fine root survival probability, but there was no difference between WL treatments. Shoot growth suffered more from ContWL and root growth more from IntWL. Needle concentrations of pinitol increased in the WL saplings, indicating stress. No WL effects were observed in photosynthesis and chlorophyll fluorescence. Increased starch concentration in needles by WL may be due to damaged roots and thus a missing belowground sink. Electrical impedance indicated suffering of WL saplings, although root hydraulic conductance did not differ between the treatments. Oxidative stress of short-term and interval WL can have long-lasting effects on shoot and root growth and the physiology of Scots pine. We conclude that even short-term WL during the growing season is a stress factor, which will probably increase in the future and can affect carbon allocation and dynamics in boreal forests.

Long generation times have been suggested to hamper rapid genetic adaptation of organisms to changing environmental conditions. We examined if environmental memory of the parental Scots pines (Pinus sylvestris L.) drive offspring survival and growth. We used seeds from trees growing under naturally dry conditions (control), irrigated trees (irrigated from 2003 to 2016), and formerly irrigated trees (“irrigation stop”; irrigated from 2003–2013; control condition since 2014). We performed two experiments, one under controlled greenhouse conditions and one at the experimental field site. In the greenhouse, the offspring from control trees exposed regularly to drought were more tolerant to hot–drought conditions than the offspring from irrigated trees and showed lower mortality even though there was no genetic difference. However, under optimal conditions (high water supply and full sunlight), these offspring showed lower growth and were outperformed by the offspring of the irrigated trees. This different offspring growth, with the offspring of the “irrigation‐stop” trees showing intermediate responses, points to the important role of transgenerational memory for the long‐term acclimation of trees. Such memory effects, however, may be overridden by climatic extremes during germination and early growth stages such as the European 2018 mega‐drought that impacted our field experiment., Environmental conditions (natural dry vs. irrigated) experienced by the mother trees of Pinus sylvestris significantly influenced the offspring's growth and survival, indicating that transgenerational environmental memory can shape trees' acclimation potential over the midterm to the long term.

SummaryScots pine (Pinus sylvestris) is the most widespread coniferous tree in the boreal forests of Eurasia and has major economic and ecological importance. However, its large and repetitive genome presents a challenge for conducting genome-wide analyses such as association studies and genomic selection. We present a new 50K SNP genotyping array for Scots pine research, breeding programs, and other applications. To select the SNP set, we first genotyped 480 Scots pine samples on a 407 540 SNP screening array, and identified 47 712 high-quality SNPs for the final array (called ‘PiSy50k’). Here, we provide details of the design and testing, as well as allele frequency estimates from the discovery panel, functional annotation, tissue-specific expression patterns, and expression level information for the SNPs or corresponding genes, when available. We validated the performance of the PiSy50k array using samples from breeding populations from Finland and Scotland. Overall, 39 678 (83.2%) SNPs showed low error rates (mean = 0.92%). Relatedness estimates based on array genotypes were consistent with the expected pedigrees, and the amount of Mendelian error was negligible. In addition, array genotypes successfully discriminate Scots pine populations from different geographic origins. The PiSy50k array will be a valuable tool for future genetic studies and forestry applications.Significance statementScots pine is an evolutionary, economically and ecologically impressive coniferous species but its gigantic genome has limited studying e.g. the genetic basis of its functional trait variation. We have developed a genotyping array that facilitates Scots pine genetic research and linking its trait variation to genetic polymorphisms and gene expression levels across the genome.

Research Highlights: The antifungal assay confirmed that knotwood extractives of Scots pine inhibit the growth of wood decay fungi. Heartwood extracts of black locust were found to be much stronger free radical scavengers than the extracts of Scots pine. The extracts were deposited in the lumina and on the wall surface of cells in the impregnated sapwood. Impregnation of the sapwood blocks with Scots pine and black locust extracts reduced the fungal decay of wood. Objectives: Hydrophilic extracts of Scots pine knotwood and black locust heartwood were chemically analyzed, tested for antifungal and antioxidant properties and used for impregnation of beech and Scots pine sapwood. Materials and Methods: Scots pine knotwood and black locust heartwood were extracted, and obtained hydrophilic extractives were chemically analyzed. Extracts were analyzed for antifungal properties with the in vitro well-diffusion method. The free radical scavenging activity of wood extracts was measured colorimetrically. The retention of the extracts in the impregnated sapwood blocks was evaluated with microscopy and gravimetry. A decay test was performed with the mini block test. Results: Almost half of both Scots pine knotwood and black locust heartwood hydrophilic extracts obtained were described by phenolic compounds. The extracts were deposited in the lumina of cells and on the cell wall surface. Extractives of Scots pine knotwood had good inhibitory properties against white- and brown-rot fungi. On the other hand, extractives of black locust heartwood were found to be good radical scavengers, better than knotwood extractives of Scots pine. The extracts of Scots pine knotwood and black locust reduced the fungal decay of the tested sapwood blocks. Conclusions: The results of this research show that the less-valued knotwood of Scots pine and heartwood of black locust are a potential source of antifungal and antioxidant agents for bio-based wood preservatives.

Microbes living in plant tissues—endophytes—are mainly studied in crop plants where they typically colonize the root apoplast. Trees—a large carbon source with a high capacity for photosynthesis—provide a variety of niches for endophytic colonization. We have earlier identified a new type of plant–endophyte interaction in buds of adult Scots pine, where Methylorubrum species live inside the meristematic cells. The endosymbiont Methylorubrum extorquens DSM13060 significantly increases needle and root growth of pine seedlings without producing plant hormones, but by aggregating around host nuclei. Here, we studied gene expression and metabolites of the pine host induced by M. extorquens DSM13060 infection. Malic acid was produced by pine to potentially boost M. extorquens colonization and interaction. Based on gene expression, the endosymbiont activated the auxin- and ethylene (ET)-associated hormonal pathways through induction of CUL1 and HYL1, and suppressed salicylic and abscisic acid signaling of pine. Infection by the endosymbiont had an effect on pine meristem and leaf development through activation of GLP1-7 and ALE2, and suppressed flowering, root hair and lateral root formation by downregulation of AGL8, plantacyanin, GASA7, COW1 and RALFL34. Despite of systemic infection of pine seedlings by the endosymbiont, the pine genes CUL1, ETR2, ERF3, HYL, GLP1-7 and CYP71 were highly expressed in the shoot apical meristem, rarely in needles and not in stem or root tissues. Low expression of MERI5, CLH2, EULS3 and high quantities of ononitol suggest that endosymbiont promotes viability and protects pine seedlings against abiotic stress. Our results indicate that the endosymbiont positively affects host development and stress tolerance through mechanisms previously unknown for endophytic bacteria, manipulation of plant hormone signaling pathways, downregulation of senescence and cell death-associated genes and induction of ononitol biosynthesis.

Tree stems have been identified as sources of volatile organic compounds (VOCs) that play important roles in tree defence and atmospheric chemistry. Yet, we lack understanding on the magnitude and environmental drivers of stem VOC emissions in various forest ecosystems. Due to the increasing importance of extreme drought, we studied drought effects on the VOC emissions from mature Scots pine (Pinus sylvestris L.) stems. We measured monoterpenes, acetone, acetaldehyde and methanol emissions with custom-made stem chambers, online PTR-MS and adsorbent sampling in a drought-prone forest over the hot-dry summer of 2018 and compared the emission rates and dynamics between trees in naturally dry conditions and under long-term irrigation (drought release). The pine stems were significant monoterpene sources. The stem monoterpene emissions potentially originated from resin, based on their similar monoterpene spectra. The emission dynamics of all VOCs followed temperature at a daily scale, but monoterpene and acetaldehyde emission rates decreased nonlinearly with drought over the summer. Despite the dry conditions, large peaks of monoterpene, acetaldehyde and acetone emissions occurred in late summer potentially due to abiotic or biotic stressors. Our results highlight the potential importance of stem emissions in the ecosystem VOC budget, encouraging further studies in diverse environments.

International audience; Climate change affects forest ecosystem processes and related services due to climate variability. These might affect ecosystem functioning, especially productivity. Regarding management issues, mixed stands are considered a relevant option to maintain forest cover and ecosystem services under climate change. However, the possibility to maintain these mixed stands with management actions with positive effects on forest functioning under climate change remains uncertain and deserves further investigations. Relying on a simulation-based study with a forest gap model (ForCEEPS), we thus addressed the following questions: (1) Are monospecific stands vulnerable to climate change? (2) Would mixed stands significantly mitigate climate change effects on forest productivity and wood production under climate change? (3) Would conversion to mixed stand management affect significantly forest productivity and wood production under climate change compare to monospecific management? In this study, we quantified potential climate change effect (using RCP 8.5 and present climate) and management’s effect in the French Alps, focusing on five species (Fagus sylvatica, Abies alba, Picea abies, Pinus sylvestris, and Quercus pubescens). We tested different scenarios, with various composition, structure, or environmental conditions, under climate change. These simulations showed that monospecific stands currently growing in stressful conditions would be vulnerable to climate change. Managing mixed stands or conversion from pure to mixed stands would make it possible to maintain higher productivity in the long term than monospecific stands, depending on the species and the sites considered. Our results will feed into discussion on forest management in the context of climate change.

Monitoring early tree physiological responses to drought is key to understanding progressive impacts of drought on forests and identifying resilient species. We combined drone-based multispectral remote sensing with measurements of tree physiology and environmental parameters over two growing seasons in a 100-y-old Pinus sylvestris forest subject to 17-y of precipitation manipulation. Our goal was to determine if drone-based photochemical reflectance index (PRI) captures tree drought stress responses and whether responses are affected by long-term acclimation. PRI detects changes in xanthophyll cycle pigment dynamics, which reflect increases in photoprotective non-photochemical quenching activity resulting from drought-induced photosynthesis downregulation. Here, PRI of never-irrigated trees was up to 10 times lower (higher stress) than PRI of irrigated trees. Long-term acclimation to experimental treatment, however, influenced the seasonal relationship between PRI and soil water availability. PRI also captured diurnal decreases in photochemical efficiency, driven by vapour pressure deficit. Interestingly, 5 years after irrigation was stopped for a subset of the irrigated trees, a positive legacy effect persisted, with lower stress responses (higher PRI) compared with never-irrigated trees. This study demonstrates the ability of remotely sensed PRI to scale tree physiological responses to an entire forest and the importance of long-term acclimation in determining current drought stress responses.

Boreal trees are capable of taking up organic nitrogen (N) as effectively as inorganic N. Depending on the abundance of soil N forms, plants may adjust physiological and morphological traits to optimize N uptake. However, the link between these traits and N uptake in response to soil N sources is poorly understood. We examined Pinus sylvestris L. seedlings’ biomass growth and allocation, transpiration and N uptake in response to additions of organic N (the amino acid arginine) or inorganic N (ammonium nitrate). We also monitored in situ soil N fluxes in the pots following an addition of N, using a microdialysis system. Supplying organic N resulted in a stable soil N flux, whereas the inorganic N resulted in a sharp increase of nitrate flux followed by a rapid decline, demonstrating a fluctuating N supply and a risk for loss of nitrate from the growth medium. Seedlings supplied with organic N achieved a greater biomass with a higher N content, thus reaching a higher N recovery compared with those supplied inorganic N. In spite of a higher N concentration in organic N seedlings, root-to-shoot ratio and transpiration per unit leaf area were similar to those of inorganic N seedlings. We conclude that enhanced seedlings’ nutrition and growth under the organic N source may be attributed to a stable supply of N, owing to a strong retention rate in the soil medium.

Compared to angiosperms, gymnosperms lag behind in the availability of assembled and annotated genomes. Most genomic analyses in gymnosperms, especially conifer tree species, rely on the use of de novo assembled transcriptomes. However, the level of allelic redundancy and transcript fragmentation in these assembled transcriptomes, and their effect on downstream applications have not been fully investigated. Here, we assessed three assembly strategies, including the utility of haploid (megagametophyte) tissue during de novo assembly as single-allele guides, for six individuals and five different tissues in Pinus sylvestris. We then contrasted haploid and diploid tissue genotype calls obtained from the assembled transcriptomes to evaluate the extent of paralog mapping. The use of the haploid tissue during assembly increased its completeness without reducing the number of assembled transcripts. Our results suggest that current strategies that rely on available genomic resources as guidance to minimize allelic redundancy are less effective than the application of strategies that cluster redundant assembled transcripts. The strategy yielding the lowest levels of allelic redundancy among the assembled transcriptomes assessed here was the generation of SuperTranscripts with Lace followed by CD-HIT clustering. However, we still observed some levels of heterozygosity (multiple gene fragments per transcript reflecting allelic redundancy) in this assembled transcriptome on the haploid tissue, indicating that further filtering is required before using these assemblies for downstream applications. We discuss the influence of allelic redundancy when these reference transcriptomes are used to select regions for probe design of exome capture baits and for estimation of population genetic diversity.

International audience; The extent to which water availability can be used to predict the enlargement and final dimensions of xylem conduits remains an open issue. We reconstructed the time course of tracheid enlargement in Pinus sylvestris trees in central Spain by repeated measurements of tracheid diameter on microcores sampled weekly during a 2 yr period. We analyzed the role of water availability in these dynamics empirically through time-series correlation analysis and mechanistically by building a model that simulates daily tracheid enlargement rate and duration based on Lockhart's equation and water potential as the sole input. Tracheid enlargement followed a sigmoid-like time course, which varied intra- and interannually. Our empirical analysis showed that final tracheid diameter was strongly related to water availability during tracheid enlargement. The mechanistic model was calibrated and successfully validated (R-2 = 0.92) against the observed tracheid enlargement time course. The model was also able to reproduce the seasonal variations of tracheid enlargement rate, duration and final diameter (R-2 = 0.84-0.99). Our results support the hypothesis that tracheid enlargement and final dimensions can be modeled based on the direct effect of water potential on turgor-driven cell expansion. We argue that such a mechanism is consistent with other reported patterns of tracheid dimension variation.

Embargo until 08 July 2020 Exposure to ambient UV-B radiation may prime protective responses towards various stressors in plants, though information about interactive effects of UV-B and gamma radiation is scarce. Here, we aimed to test whether UV-B exposure could prime acclimatisation mechanisms contributing to tolerance to low-moderate gamma radiation levels in Scots pine seedlings, and concurrently whether simultaneous UV-B and gamma exposure may have an additive adverse effect on seedlings that had previously not encountered either of these stressors. Responses to simultaneous UV-B (0.35 W m−2) and gamma radiation (10.2–125 mGy h−1) for 6 days with or without UV-B pre-exposure (0.35 W m−2, 4 days) were studied across various levels of organisation, as compared to effects of either radiation type. In contrast to UV-B, and regardless of UV-B presence, gamma radiation at ≥42.9 mGy h−1 caused increased formation of reactive oxygen species and reduced shoot length, and reduced root length at 125 mGy h−1. In all experiments there was a gamma dose rate-dependent increase in DNA damage at ≥10.8 mGy h−1, generally with additional UV-B-induced damage. Gamma-induced growth inhibition and gamma- and UV-B-induced DNA damage were still visible 44 days post-irradiation, even at 20.7 mGy h−1, probably due to genomic instability, but this was reversed after 8 months. In conclusion, there was no evidence of a protective effect of UV-B on gamma-induced growth inhibition and DNA damage in Scots pine, and no additive adverse effect of gamma and UV-B radiation on growth in spite of the additional UV-B-induced DNA damage.

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.

Pinus sylvestris has a long history of basic and applied research that is relevant for both forestry and evolutionary studies. Its patterns of adaptive variation and role in forest economic and ecological systems have been studied extensively for nearly 275 years, detailed demography for a 100 years and mating system more than 50 years. However, its reference genome sequence is not yet available and genomic studies have been lagging compared to, for example, Pinus taeda and Picea abies, two other economically important conifers. Despite the lack of reference genome, many modern genomic methods are applicable for a more detailed look at its biological characteristics. For example, RNA‐seq has revealed a complex transcriptional landscape and targeted DNA sequencing displays an excess of rare variants and geographically homogenously distributed molecular genetic diversity. Current DNA and RNA resources can be used as a reference for gene expression studies, SNP discovery, and further targeted sequencing. In the future, specific consequences of the large genome size, such as functional effects of regulatory open chromatin regions and transposable elements, should be investigated more carefully. For forest breeding and long‐term management purposes, genomic data can help in assessing the genetic basis of inbreeding depression and the application of genomic tools for genomic prediction and relatedness estimates. Given the challenges of breeding (long generation time, no easy vegetative propagation) and the economic importance, application of genomic tools has a potential to have a considerable impact. Here, we explore how genomic characteristics of P. sylvestris, such as rare alleles and the low extent of linkage disequilibrium, impact the applicability and power of the tools.

Litterfall dynamics (production, seasonality and nutrient composition) are key factors influencing nutrient cycling. Leaf litter characteristics are modified by species composition, site conditions and water availability. However, significant evidence on how large-scale, global circulation patterns affect ecophysiological processes at tree and ecosystem level remains scarce due to the difficulty in separating the combined influence of different factors on local climate and tree phenology. To fill this gap, we studied links between leaf litter dynamics with climate and other forest processes, such as tree-ring width (TRW) and intrinsic water-use efficiency (iWUE) in two mixtures of Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) in the south-western Pyrenees. Temporal series (18 years) of litterfall production and elemental chemical composition were decomposed following the ensemble empirical mode decomposition (EEMD) method and relationships with local climate, large-scale climatic indices, TRW and Scots pine´s iWUE were assessed. Temporal trends in N:P ratios indicated increasing P-limitation of soil microbes, thus affecting nutrient availability, as the ecological succession from a pine-dominated to a beech-dominated forest took place. A significant influence of large-scale patterns on tree-level ecophysiology was explained through the impact of the North Atlantic Oscillation (NAO) and El Niño – Southern Oscillation (ENSO) on water availability. Positive NAO and negative ENSO were related to dry conditions and, consequently, to early needle shedding and increased N:P ratio of both species. Autumn storm activity appears to be related to premature leaf abscission of European beech. Significant cascading effects from large-scale patterns on local weather influenced pine TRW and iWUE. These variables also responded to leaf stoichiometry fallen three years prior to tree-ring formation. Our results provide evidence of the cascading effect that variability in global climate circulation patterns can have on ecophysiological processes and stand dynamics in mixed forests. Ministerio de Economía y Competitividad, Grant/Award Number: AGL2012-33465, AGL2016-76463-P, BES-2013-066705 and RYC-2011-08082; FP7 People: Marie-Curie Actions, Grant/Award Number: CIG-2012-326718-ECOPYREN3; H2020 Marie Skłodowska-Curie Actions, Grant/Award Number: EF-2014-656810-DENDRONUTRIENT.

In this study we derived allometric functions for estimating the belowground biomass (BGB) of Silver Birch (Betula pendula Roth), Pedunculate Oak (Quercus robur L.), Sessile Oak (Quercus petraea (Matt.) Liebl.) and Scots Pine (Pinus sylvestris L.) in Germany. To assess the impact on German greenhouse gas (GHG) reporting, these new functions were further compared with BGB functions currently used in France and Sweden. For developing new BGB functions 48 Silver Birches, 39 Pedunculate and Sessile Oaks and 54 Scots Pines were destructively sampled. The sampled trees spanned a DBH range from 8.2 to 52.9 cm for Silver Birch, from 7.4 to 42.0 cm for Oak and from 7.2 to 53.2 cm for Scots Pine. After fitting the data, the following values of model efficiency were achieved: 0.81 for Silver Birch, 0.98 for Oak and 0.95 for Scots Pine. The model root mean square error varies between 5.2 kg for Oak, 13.7 kg for Scots pine and 26.9 kg for Silver Birch. Comparison with the currently applied BGB functions in the German GHG inventory from France and Sweden showed that the use of these functions results in systematically different estimates for the BGB of Silver Birch and Oak. Thus, our findings indicate that BGB functions recommended for other European countries (in particular France and Sweden) are not appropriate for estimating the BGB for the tree species concerned in Germany. Currently, the derived data-set for BGB of Silver Birch, Oak and Scots Pine is the largest in Germany and the developed functions are thus the best available for estimating national BGB stock and stock change in Germany at the moment.

Soil waterlogging is predicted to increase in the future climate in boreal regions due to increased precipitation. Snowmelt periods in winter may also become more common and further increase the amount of water in soil. It is not well known how waterlogging and soil freezing during winter affect the physiology, phenology and growth of trees. Our aim was to study the below- and aboveground responses of Scots pine (Pinus sylvestris L.) saplings to waterlogging (WL) in frozen (Fr) and unfrozen (NoFr) soils in a growth chamber experiment. The soil was either -2 °C or +2 °C and either waterlogged or not in a split-plot design for 6 weeks during dormancy, with similar air conditions in all treatments, which were Fr + WL, NoFr + WL, Fr + NoWL and NoFr + NoWL. Needles showed a shift towards a deeper dormancy in frozen than unfrozen soil in terms of chlorophyll fluorescence (Fv/Fm), water potential and apoplastic electrical resistance. In spring, initiation of shoot elongation started earlier if the soil was frozen during dormancy. In Fr + WL, initiation of root growth was delayed by 20 days compared with other treatments; after that, the root growth peaked at the same time as needle elongation. Needles remained smaller in Fr + WL than in the other treatments, indicating that roots formed a strong sink for carbon. Shoot and root biomass were not negatively affected by waterlogging if the soil remained unfrozen. In Fr + WL, survival and growth capacity of new terminal and whorl buds, the number of bud scales and the number of dwarf shoots were reduced. We conclude that soil freezing on sites prone to waterlogging should be considered in management of boreal forests, especially in the face of predicted climate change.

International audience; National Forest Inventories (NFIs) perform systematic forest surveys across space and time. They are hence powerful tools to understand climate controls on forest growth at wide geographical scales and account for the effects of local abiotic and biotic interactions. To investigate the effects of climate change upon growth dynamics of four major European conifer species along elevation and continentality gradients, we herein provide an original harmonization of the French and Austrian NFI datasets. The growth of Norway spruce, Scots pine, silver fir and European larch over the 1996-2016 period was studied in pure and evenaged plots across different ecological regions. We derived climate-driven growth trends from > 65, 000 radial increment series filtered-out from major biotic and abiotic influences using statistical modeling. We further identified primary environmental drivers of conifer growth by regressing growth trends against regionally-aggregated biotic and abiotic forest attributes. Negative growth trends were observed in continental regions undergoing the most rapid warming and thermal amplitude contraction over the study period. Negative trends were also associated with lower forest structural heterogeneity and, surprisingly, with greater available water capacity. Remarkably, we observed these associations both at the inter-and intraspecies levels, suggesting the universality of these primary growth determinants. Our study shows that harmonized NFI data at the transnational level provide reliable information on climate-growth interactions. Here, greater forest structural complexity and greater water resource limitation were highlighted as drivers of greater forest resilience to climate change at large-scale. This result forms crucial bases to implementing climate-smart forest management.

In temperate European forests invaded by Prunus serotina Ehrh. (black cherry), a reduction in the spontaneous regeneration capacity of Pinus sylvestris L. (Scots pine) is observed. It could be caused by various factors, including allelopathic properties of this invasive plant. In this study the phytotoxic effect of P. serotina volatile compounds on P. sylvestris and the seasonal variation in this effect were assessed. Simple assays showed that volatiles emitted from P. serotina leaves significantly inhibited root growth of P. sylvestris seedlings. Their negative effect on stem growth was much weaker. The strongest phytotoxic effect on Scots pine seedlings was caused by the volatiles emitted from the youngest black cherry leaves. In fresh foliage of P. serotina, nineteen volatile organic compounds were identified by gas chromatography–mass spectrometry (GC–MS). The dominant compound was benzaldehyde. On the basis of tests of linalool alone, it was found that this monoterpene present in the volatile fraction has a strong allelopathic potential and inhibits germination, root elongation and shoot elongation of pine seedlings. The results of our research suggest that volatile compounds from P. serotina leaves could limited survival of P. sylvestris individuals in the seedling phase.

Publisher Copyright: © 2020 The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. The heartwood of many wood species is rich in extractives, which improve the wood material's resistance to biological attack. Their concentration is generally higher in outer than inner heartwood, but the exact radial changes in aging heartwood remain poorly characterized. This investigation studied these radial changes in detail in Scots pine (Pinus sylvestris L.), using radial sample sequences prepared from three different trees. Stilbene and resin acid contents were first measured from bulk samples, after which the extractive contents of individual heartwood annual rings were investigated using Raman spectroscopy and fluorescence microscopy. Raman imaging and fluorescence microscopy were also used to study the cellular-level distributions of extractives in different annual rings. Although there were substantial differences between the trees, the content and distribution of stilbenes seemed to follow a general radial trend. The results suggest that stilbenes are absorbed into heartwood tracheid cell walls from small stilbene-rich extractive deposits over several years and then eventually transform into non-extractable compounds in aging heartwood. Resin acids followed no consistent radial trends, but their content was strongly connected to the frequency of large extractive deposits in latewood tracheid lumens. The results highlight the variability of heartwood extractives: their content and distribution vary not only between trees but also between and even within the annual rings of a single tree. This high variability is likely to have important effects on the properties of heartwood and the utilization of heartwood timber.

Supplementary Materials: The following are available online at https://www.mdpi.com/article/ 10.3390/fire4020029/s1, Paleoecological Methods; Table S1: radiocarbon (AMS-14C) data from Arroyo de Aguas Frías site, Mountain forests are subjected to several pressures including historical land-use changes and climate warming which may lead to shifts in wildfire severity negatively impacting tree species with low post-fire growth resilience. This is the case of relict Mediterranean Scots pine (Pinus sylvestris) forests in the Sierra de Gredos mountains (central Spain). We reconstructed the historical fire regime of these forests since 1700 by using paleoecology, historical ecology and dendroecology. We detected an increase in charcoal accumulation rate and coprophilous fungi in peat bogs during the late 19th century when the pine pollen percentage sharply decreased, historical records of fire peaked and many trees showed growth suppressions. We inferred an increased wildfire incidence during the late 19th century, which could have shaped the current distribution of Scots pine forests. This shift in fire-forest interactions can be explained by the uncontrolled use of mountain forests and grasslands due to the dissolution of “Mesta”, one of the major and lasting transhumance livestock associations in Europe. Integrating historical human and climate influences on fire regimes allows decomposing the resilience and conservation components of relict forests

After drought-induced embolism and repair, tree xylem may be weakened against future drought events (cavitation fatigue). As there are few data on cavitation fatigue in conifers available, we quantified vulnerability curves (VCs) after embolism/repair cycles on eight European conifer species. We induced 50% and 100% loss of conductivity (LC) with a cavitron, and analyzed VCs. Embolism repair was obtained by vacuum infiltration. All species demonstrated complete embolism repair and a lack of any cavitation fatigue after 50% LC . After 100% LC, European larch (Larix decidua), stone pine (Pinus cembra), Norway spruce (Picea abies), and silver fir (Abies alba) remained unaffected, while mountain pine (Pinus mugo), yew (Taxus baccata), and common juniper (Juniperus communis) exhibited 0.4–0.9 MPa higher vulnerability to embolism. A small cavitation fatigue observed in Scots pine (Pinus sylvestris) was probably biased by incomplete embolism repair, as indicated by a correlation of vulnerability shifts and conductivity restoration. Our data demonstrate that cavitation fatigue in conifers is species-specific and depends on the intensity of preceding LC. The lack of fatigue effects after moderate LC, and relevant effects in only three species after high LC, indicate that conifers are relatively resistant against cavitation fatigue. This is remarkable considering the complex and delicate conifer pit architecture and may be important considering climate change projections.

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.

Early-stage fitness variation has been seldom evaluated at broad scales in forest tree species, despite the long tradition of studying climate-driven intraspecific genetic variation. In this study, we evaluated the role of climate in driving patterns of population differentiation at early-life stages in Pinus sylvestris and explored the fitness and growth consequences of seed transfer within the species range. We monitored seedling emergence, survival and growth over a 2-yr period in a multi-site common garden experiment which included 18 European populations and spanned 25 degrees in latitude and 1700 m in elevation. Climate-fitness functions showed that populations exhibited higher seedling survival and growth at temperatures similar to their home environment, which is consistent with local adaptation. Northern populations experienced lower survival and growth at warmer sites, contrary to previous studies on later life stages. Seed mass was higher in populations from warmer areas and was positively associated with survival and growth at more southern sites. Finally, we did not detect a survival-growth trade-off; on the contrary, bigger seedlings exhibited higher survival probabilities under most climatic conditions. In conclusion, our results reveal that contrasting temperature regimes have played an important role in driving the divergent evolution of P. sylvestris populations at early-life stages.

Understanding the dynamics of selection is key to predicting the response of tree species to new environmental conditions in the current context of climate change. However, selection patterns acting on early recruitment stages and their climatic drivers remain largely unknown in most tree species, despite being a critical period of their life cycle. We measured phenotypic selection on Pinus sylvestris seed mass, emergence time and early growth rate over 2 yr in four common garden experiments established along the latitudinal gradient of the species in Europe. Significant phenotypic plasticity and among-population genetic variation were found for all measured phenotypic traits. Heat and drought negatively affected fitness in the southern sites, but heavy rainfalls also decreased early survival in middle latitudes. Climate-driven directional selection was found for higher seed mass and earlier emergence time, while the form of selection on seedling growth rates differed among sites and populations. Evidence of adaptive and maladaptive phenotypic plasticity was found for emergence time and early growth rate, respectively. Seed mass, emergence time and early growth rate have an adaptive role in the early stages of P. sylvestris and climate strongly influences the patterns of selection on these fitness-related traits., This study was funded by the European Union Horizon 2020 research and innovation program under grant agreement no.676876 (GenTree project). ASM was supported by a PhD fellowship (FPI-SGIT-INIA) and TP by the Academy of Finland (287431). We are greatly indebted to all GenTree partner teams that participated in the seed collection campaigns: NIBIO,NERC, CNR, WSL, INRA and THUNEN. Finnish seeds were provided by Natural Resources Institute Finland, (LUKE).

Harsh environmental conditions affect both leaf structure and root traits. However, shoot growth in high-latitude systems is predominately under photoperiod control while root growth may occur for as long as thermal conditions are favorable. The different sensitivities of these organs may alter functional relationships above- and belowground along environmental gradients. We examined the relationship between absorptive root and foliar traits of Scots pine trees growing in situ along a temperate-boreal transect and in trees grown in a long-term common garden at a temperate latitude. We related changes in foliar nitrogen, phosphorus, specific leaf area, needle mass and 13 C signatures to geographic trends in absorptive root biomass to better understand patterns of altered tree nutrition and water balance. Increased allocation to absorptive fine roots was associated with greater uptake of soil nutrients and subsequently higher needle nutrient contents in the northern provenances compared with more southern provenances when grown together in a common garden setting. In contrast, the leaf δ13 C in northern and southern provenances were similar within the common garden suggesting that higher absorptive root biomass fractions could not adequately increase water supply in warmer climates. These results highlight the importance of allocation within the fine-root system and its impacts on needle nutrition while also suggesting increasing stomatal limitation of photosynthesis in the context of anticipated climatic changes.

Summary The effect of drought on the interplay of processes controlling carbon partitioning into plant primary and secondary metabolisms, such as respiratory CO2 release and volatile organic compound (VOC) biosynthesis, is not fully understood. To elucidate the effect of drought on the fate of cellular C sources into VOCs vs CO2, we conducted tracer experiments with 13CO2 and position-specific 13C-labelled pyruvate, a key metabolite between primary and secondary metabolisms, in Scots pine seedlings. We determined the stable carbon isotope composition of leaf exchanged CO2 and VOC. Drought reduced the emission of the sesquiterpenes α-farnesene and β-farnesene but did not affect 13C-incorporation from 13C-pyruvate. The labelling patterns suggest that farnesene biosynthesis partially depends on isopentenyl diphosphate crosstalk between chloroplasts and cytosol, and that drought inhibits this process. Contrary to sesquiterpenes, drought did not affect emission of isoprene, monoterpenes and some oxygenated compounds. During the day, pyruvate was used in the TCA cycle to a minor degree but was mainly consumed in pathways of secondary metabolism. Drought partly inhibited such pathways, while allocation into the TCA cycle increased. Drought caused a re-direction of pyruvate consuming pathways, which contributed to maintenance of isoprene and monoterpene production despite strongly inhibited photosynthesis. This underlines the importance of these volatiles for stress tolerance., This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (ERC consolidator grant VOCO2 #647008). The authors are also grateful to Monika Eiblmeier for technical assistance in GC-MS/IRMS analyses, Michael Rienks for logistical help and Manizha Khakimova for gardening the plants.

Changes in the climate may have unpredictable effects on belowground carbon processes and thus, the carbon balance of boreal forests. To understand the interactions of these processes in soil and to quantify the potential changes in the carbon cycle, partitioning of forest floor respiration is crucial. For this purpose, we used nine different treatments to separate the sources of forest floor carbon dioxide (CO2) emissions in a mature Scots pine (Pinus sylvestris L.) stand in southern Finland. To partition the belowground CO2 emissions, we used two different trenching methods: 1) to exclude roots and mycorrhizal fungal mycelia (mesh with 1-µm pores) and 2) to exclude roots, but not mycorrhizal hyphae (mesh with 50-µm pores). Additionally, we used 3) a control treatment that included roots and fungal hyphae. To partition the CO2 emissions from the forest floor vegetation, we 1) removed it, 2) left only the dwarf shrubs, or 3) left the vegetation intact. The forest floor CO2 emissions were regularly measured with a flux chamber throughout the growing seasons in 2013–2015. The total forest floor respiration was partitioned into respiration of tree roots (contributing 48%), heterotrophic soil respiration (30%) and respiration of ground vegetation other than shrubs (10%), dwarf shrubs (8%), and hyphae of mycorrhizal fungi (4%). Heterotrophic respiration increased in the trenched treatments without ground vegetation over time, due to the so-called ‘Gadgil effect’. In the absence of tree roots, but when hyphal access was allowed, respiration in the dwarf shrub treatment increased throughout the experiment. This indicated that dwarf shrubs had fungal connections to outside the experimental plots via their ericoid mycorrhiza. At the same time, other ground vegetation, such as mosses, suppressed the dwarf shrub respiration in trenched treatments. Our results show that competition on the forest floor is intense between plant roots and soil microbes. publishedVersion

The bark resulted after the industrial processing of wood represents a byproduct of the forestry industry, used in many suboptimal ways, being considered a natural waste. Currently, it has been highlighted that the bark of different woody plants may be an important source of several bioactive compounds, with various beneficial biological functions. The aim of this study is to evaluate and compare the chemical composition and biological activities of two different pine species (Pinus nigra and Pinus sylvestris) bark extracts. Ultrasound (UAE) and microwave (MAE) assisted extractions were performed in order to obtain the extracts. The total polyphenol (TPC) and total tannin (TTC) contents of the extracts were assessed via the Folin–Ciocâlteu method. The volatile and polyphenolic compounds were identified and quantified via a GC/MS analysis and an UPLC-PDA analysis, respectively. The antioxidant activity (AOA) was evaluated using the DPPH and ABTS assays, while the antibacterial activity was assessed using the minimum inhibitory concentration (MIC) protocol. The results showed that the Pinus sylvestris bark extracts obtained by UAE had a higher TPC, TTC and AOA, while the Pinus nigra bark extracts obtained by MAE had higher volatile compounds content (mainly α-pinene and β-pinene). Moreover, the inhibition of the bacterial growth was more efficient in the case of Pinus sylvestris extracts, Gram-positive bacteria being the most affected, while Gram-negative strains presented a relative resistance to the tested extract concentrations. These results may indicate the potential use of the pine bark extracts as antioxidant or antibacterial agents.

Phoretic mites use bark beetles for transportation to new, suitable habitats. Some phoretic mites act as predators and parasitoids of the bark beetles’ immature stages, especially egg and early larval stages, and are potential agents for the biological control of scolytine forest pests. Mites live very frequently in relationships with other invertebrates. Many are found in association with various species of bark beetles. Here, a total of 41 specimens of different bark beetles of the genus Ips (Ips acuminatus, Ips sexdentatus and Ips typographus) were studied for presence, species composition, and abundance of phoretic mites. The beetles were collected on dead wood and parts of tree bark of Pinus nigra, Pinus sylvestris and Picea abies in the Black Sea Mountains in Kastamonu and Artin Province of Turkey. A total of nine mite species were found, including Dendrolaelaps quadrisetus, Ereynetes sp., Histiostoma piceae, Paraleius cf. leontonychus, Pleuronectocelaeno barbara., Proctolaelaps hystricoides, Schizostethus simulatrix, Trichouropoda lamellosa and Uroobovellaipidis. All species are identified for the first time within Turkish fauna.

In recent years, large predators have made a comeback across large parts of Europe. However, little is known about the impact that recolonizing predators may have on ecosystems with high degrees of anthropogenic influence. In Scandinavia, wolves (Canis lupus) now inhabit areas affected by intense forestry practices and their main prey, moose (Alces alces), are exposed to significant human hunting pressure. We used long-term datasets to investigate whether the return of wolves has affected moose distribution (i.e., presence and abundance) as well as browsing damage (i.e., presence and intensity) by moose on Scots pine (Pinus sylvestris). We found that the probability of moose presence and abundance increased with time since wolf territory establishment and was higher inside wolf territories than outside. Additionally, the probability of browsing damage was also higher inside wolf territories compared to outside, but wolf occurrence had no effect on browsing damage intensity. We suggest two possible underlying mechanisms behind these results: (1) wolves might select to establish territories in areas with higher moose abundance, increasing their probability of encounters, and/or (2) hunters within wolf territories reduce the number of harvested moose to compensate for wolf predation. This study highlights that the return of large predators to landscapes with strong anthropogenic influence may result in alternative effects than those described in studies on trophic cascades located in protected areas.

The development of mobile optoelectronic graders for separating viable seeds by spectrometric properties with high detection accuracy represents a very relevant direction of development for seed handling operations. Here, the main parameters of the radiation source and receiver for detecting a single seed in the diagnostic system of a mobile grader were modeled based on the principles of technical optics using Scots pine (Pinus sylvestris L.) seeds as a case study. Among the pine seeds in the seed batch, there are fossilized and empty seeds that are exactly the same in geometric and gravitational parameters as live seeds. For their separation from the seed batch, data from spectrometric studies in the near-infrared (980 nm) region can be used. To substantiate the parameters of the light source, a geometric optical model of optical beam formation was considered, while the energy model of optical beam formation was considered to substantiate the parameters of the light detector. The results of this study show that the signal value depended on the orientation of a single seed relative to the recording window. The beam angle from the radiation source should be within 45 degrees. The difference between the optical streams should be 50 microns, which made it possible to clearly detect the signal at a standard noise level of 15 microns and signal-to-noise detection accuracy ratio of 56.3 dB. This study expands theoretical knowledge in the field of the spectrometric properties of a single seed, considering the cases of its orientation relative to the optical beam, which affected the effective area of detection of the seed. The obtained data on the location of the main elements of the diagnostic system will speed up the design of mobile optoelectronic graders, and the development of a contemporary protocol for improving Scots pine seed quality.

Scots pine (Pinus sylvestris L.) is the most widely distributed pine species in Europe and is relevant in terms of planted areas and harvest yields. Therefore, each year the demand for planting stock of Scots pine is exceedingly high, and large quantities of seedlings are produced annually throughout Europe to carry out reforestation and afforestation programs. Abundant and diverse ectomycorrhizal (ECM) symbiosis is critical for the success of seedlings once planted in the field. To improve our knowledge of ECM fungi that inhabit bare-root nursery stock of Scots pine and understand factors that influence their diversity, we studied the assemblages of ECM fungi present across 23 bare-root forest nurseries in Poland. Nursery stock samples were characterized by a high level of ECM colonization (nearly 100%), and a total of 29 ECM fungal taxa were found on 1- and 2-year-old seedlings. The diversity of the ECM community depended substantially on the nursery and age of the seedlings, and species richness varied from 3&ndash, 10 taxa on 1-year-old seedlings and 6&ndash, 13 taxa on 2-year-old seedlings. The ECM fungal communities that developed on the studied nursery stock were characterized by the prevalence of Ascomycota over Basidiomycota members on 1-year-old seedlings. All ecological indices (diversity, dominance, and evenness) were significantly affected by age of the seedlings, most likely because dominant ECM morphotypes on 1-year-old seedlings (Wilcoxina mikolae) were replaced by other dominant ones (e.g., Suillus luteus, Rhizopogon roseolus, Thelephora terrestris, Hebeloma crustuliniforme), mostly from Basidiomycota, on 2-year-old seedlings. Across all nurseries, negative correlations were identified for diversity metrics and soil N or C, indicating that mineral and organic fertilization contributes to the differences in the ECM fungal communities in nurseries. We discuss the ecological and practical implications of the composition and diversity of ECM fungi occurring on bare-root planting stock of Scots pine.

Tree species-mixing has been suggested as one option to counteract the adverse effects of global change on tree mineral nutrition, yet the effect of mixing on nutrient availability remains poorly documented. We therefore analyzed the current foliar nutrient (N, P, K, Ca, Mg) quantities and ilr balances (isometric log transformed ratios between elements or groups of elements) for 261 European beech and 248 Scots pine trees from 15 sites, each consisting of one beech-pine mixed stand and the respective monocultures, across a gradient of environmental conditions in Europe. We hypothesized an overall positive effect of mixing on tree foliar nutrient content, and that this mixing effect would be stronger on nutrient-poor sites. Using linear mixed models and multivariate linear regression models, we first tested for the effects of species (beech/pine) and composition (pure/mixed) across all sites; we then investigated whether the species-mixing effect was related to site fertility. The nutrient composition of beech leaves and pine needles differed significantly for all ilr balances. For both species, significant mixing effects were detected for some nutrients and ilr balances; those effects, however, could not be consistently related to contrasted nutrient composition between species. For most nutrients and ilr balances, the mixing effect was influenced by the site nutritional status, but the pattern differed from expectation: absence or minor differences between monocultures and mixtures at the lower end of the chemical fertility gradient, and maximum differences in rich soils. The contrasting foliar nutrient composition of pine and beech trees and the site nutrient status only partly explained the mixing effects on tree mineral nutrition. Our results claim for a better understanding of nutrient-related mechanisms associated with complementarity and points towards the need to further expand the existing frameworks to account for the multivariate nature of tree nutrition., The main autor obtained a PhD grant from the ‘Fonds National de la Recherche Scientifique’ (FNRS-FRIA) and additional funding support from the Walloon forest service (Service Public de Wallonie—Département de la Nature et des Forêts) through the 5 year research programme “Accord cadre de recherches et de vulgarisation forestières”.

Wildfire shapes the structure, dynamic and functioning of boreal forests. With predicted warmer and drier summers, increased incidence and intensity of crown-fires may affect plant-soil interactions with consequences for post-fire fertility and forest productivity. We assessed how severity of crown- and ground-fire in boreal pine forests affected post-fire responses of soil fungal communities and their associated enzyme activities, and how variation in fire severity interacts with salvage (post-fire) logging in impacting soil fungi. Crown fire-induced tree mortality had a stronger impact on fungal biomass and community composition than did ground-fire-induced loss of soil organic matter. Severe crown-fire led to replacement of ectomycorrhizal- and litter-associated fungi by stress-tolerant ascomycetes. Elevated activities of hydrolytic enzymes in burned areas were correlated with root-associated ascomycetes and moulds, suggesting opportunistic exploitation of labile organic substrates. Fire did not, however, increase the abundance of more potent basidiomycete decomposers in the organic layer, nor did it enhance organic matter oxidation by fungal peroxidases, indicating that the potential for major post-fire losses of carbon due to stimulated decomposition is limited. Rather, peroxidase activity was low in burned areas, likely reflecting the absence of ectomycorrhizal fungi. Post-fire salvage logging induced larger shifts in fungal communities in areas with low crown-fire severity. Synthesis. Historically, boreal pine forests have been shaped by low-severity ground-fires. Our study highlights a risk that increasing occurrence of high-severity crown-fire as climate warms will have detrimental effects on mycorrhizal-mediated functions that are pivotal for maintaining organic matter turnover, soil fertility and forest resilience.

Questions Changes in climate and herbivory pressure affect northern alpine ecosystems through woody plant encroachment, altering their composition, structure and functioning. The encroachment often occurs at unequal rates across heterogeneous landscapes, hinting at the importance of habitat‐specific drivers that either hamper or facilitate woody plant establishment. Here, we assess: (1) the invasibility of three distinct alpine plant community types (heath, meadow and Salix shrubland) by Pinus sylvestris (Scots pine); and (2) the relative importance of biotic (above‐ground interactions with current vegetation, herbivory and shrub encroachment) and microclimate‐related abiotic (soil temperature, moisture and light availability) drivers of pine seedling establishment success. Location Dovrefjell, Central Norway. Methods We conducted a pine seed sowing experiment, testing how factorial combinations of above‐ground removal of co‐occurring vegetation, herbivore exclusion and willow transplantation (simulated shrub encroachment) affect pine emergence, survival and performance (new stem growth, stem height and fraction of healthy needles) in three plant communities, characteristic of alpine tundra, over a period of five years. Results Pine seedling emergence and survival were similar across plant community types. Herbivore exclusion and vegetation removal generally increased pine seedling establishment and seedling performance. Within our study, microclimate had minimal effects on pine seedling establishment and performance. These results illustrate the importance of biotic resistance to seedling establishment. Conclusion Pine seedlings can easily establish in alpine tundra, and biotic factors (above‐ground plant interactions and herbivory) are more important drivers of pine establishment in alpine tundra than abiotic, microclimate‐related, factors. Studies aiming to predict future vegetation changes should thus consider local‐scale biotic interactions in addition to abiotic factors. © 2020 The Authors. Journal of Vegetation Science published by John Wiley & Sons Ltd on behalf of International Association for Vegetation Science This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.