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In the European Alps, mean temperature has risen by 2.5 K since the end of the nineteenth century. A 2 K warming of the growing season has taken place in the last 4 decades only. The 2.5 K warming should rise the position of the climatic treeline by about 400 m. Actual shifts in uppermost tree positions reported here for the Austrian Defereggen Valley and the Swiss Lower Engadine region of the Eastern Alps reach only around 140 m of elevation above the limit of old trees that date back to the nineteenth century. Uppermost Pinus cembra trees of > 2 m height currently occur at c. 2500 m, representing elevation records for the Eastern Alps. In situ temperature records for 2022–2023 revealed seasonal mean temperatures for uppermost trees that are 1–3 K higher than the equilibrium treeline isotherm of c. 6 °C in both regions (corrected for temperature anomalies from long-term records). The 2 K span reflects microhabitat differences and two ways to define the season. Thus, tree advances lag behind the upslope shift of the treeline isotherm, on average, by more than 200 m. The uppermost trees currently grow under quite warm conditions with annual shoot length increments frequently reaching 20 cm. Even without additional future warming, the new steady-state climatic treeline will exceed the Holocene maximum elevation in the Eastern Alps substantially. [ABSTRACT FROM AUTHOR]

Seed dispersal is a key component in a plant's life cycle, allowing the colonization of new habitat and eventually enabling range shifts. For animal‐dispersed plants, the behaviour of seed‐dispersing animals has direct consequences for the fate of a seed. However, there is little empirical evidence on how individual behaviour of animals affects seed dispersal.Here we investigated the effects of individual behaviour on seed dispersal distance and location by analysing movement data of spotted nutcrackers (Nucifraga caryocatactes) during harvest and scatter‐hoarding of Swiss stone pine (Pinus cembra) seeds. To assess individual variation in seed dispersal patterns, we compared daytime hourly displacement patterns across individuals. We further identified seed caching locations of individual birds and analysed their potential for pine regeneration.We found that nutcrackers showed one of two distinct behaviours: short‐distance seed dispersal and long‐distance seed dispersal. The caching locations resulting from the two behaviours differed markedly in the regeneration potential of Swiss stone pine: seeds transported over short distances were cached at high elevations with high regeneration potential within the pine's current habitat, while seeds transported over long distances were cached at low elevations with low regeneration potential outside of the pine's habitat. The behaviours were not associated with bird body mass, relative wing length or sex.Our results emphasize that intraspecific individual behaviours of seed‐dispersing animals can have far‐reaching consequences for ecosystem functions. As a major determinant of seed dispersal dynamics, individual behaviour should be taken into account for a better understanding of ecosystem functions under current and future environmental conditions. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

The heartwood of the Swiss Stone Pine, Pinus cembra L., has been scarcely investigated for secondary metabolites for a long period of time. Considering age and relative simplicity of heartwood investigations dating back to the 1940s to 1960s, we conducted the first investigation of P. cembra heartwood by HPLC, using UHPLC-DAD-ESI-MSn and HPLC-DAD techniques in combination with isolation and NMR spectroscopy, with focus on stilbenes, bibenzyls and flavonoids. Analytical problems in the HPLC analysis of Pinus stilbenes and flavonoids on reversed stationary phases were also challenged, by comparing HPLC on pentafluorophenyl (PFP) and C18 stationary phases. Seven flavonoids (1, 2, 3, 7, 8, 11, 12), four stilbenes (4, 6, 10, 13), two bibenzyls (5, 9), three fatty acids (14, 16, 17) and one diterpenic acid (15) were detected in an ethanolic extract of Pinus cembra heartwood. HPLC comparison of reversed stationary phases in HPLC showed that the antifungal, antibacterial and chemosensitizing dihydropinosylvin monomethyl ether (9) and pinosylvin monomethyl ether (10) can be separated on PFP, but not on C18 material, when eluting with a screening gradient of 20–100% acetonitrile. Flavonoid separation showed additional benefits of combining analyses on different stationary phases, as flavonoids 7 and 8 could only be separated on one of two C18 stationary phases. Earlier phytochemical results for heartwood investigations were shown to be mostly correct, yet expandable. Substances 5 to 12 were found in alignment with these references, proving remarkable phytochemical analyses at the time. Evidence for the described presence of pinobanksin could not be found. Substances 1 to 4 and 13 have to our knowledge not yet been described for P. cembra. [ABSTRACT FROM AUTHOR]

Swiss pine (Pinus cembra L.) is a montane Carpathian (Alpine-Carpathian) species. The modern distribution of the Swiss pine is disjunct, the two isolated areas are located in western and central Europe (Alps) and the middle and high mountains of the Carpathians. The largest areas of Swiss pine in Ukraine are preserved in the Gorgany region at the altitude of 1250‒1450 m above sea level, where it forms the upper forest line. In the Gorgany Nature Reserve, P. cembra grows in the altitude range from 1070 to 1475 m ASL. The lower boundary, in the form of individual trees, descends to 1000 m ASL, and the upper boundary rises to 1585 m ASL. To define the ecological preferences of the Swiss pine, a series of route surveys were conducted and research sites (permanent forest monitoring plots, stationary research profiles, and stationary areas for the counting of natural forest regeneration) were established. The study of the plant communities with P. cembra on the southwestern slopes of the Dzhurdzhi tract showed the dependence of the forest species composition on the altitude above sea level. The forest stands quantitative composition and productivity are also significantly influenced by slope exposure. The proportion of P. cembra in the forests on steep slopes (30‒55°) of west and north-west exposure is relatively high (from 17.1% to 49.0%). Under other conditions, the proportion of P. cembra in the stand composition gradually decreases. Swiss pine is almost completely absent on the slopes of eastern and northern exposures except for rare single trees in Pinus mugo Turra scrub on northern slopes. The forests with P. cembra are typical of sunny slopes where the available sum of active temperatures ranges between 1000° and 1400° and the average ambient temperatures from -12.2°С to +16°С. The optimal environmental conditions for the relict species P. cembra in the Dovbushanski Gorgany region are on the western slopes with a steepness of 17‒55° and southwestern and northwestern slopes with a steepness of 35°‒42°. In terms of soil fertility, oligotrophic edaphotopes with moderate moisture are optimal for the growth of Swiss pine. [ABSTRACT FROM AUTHOR]

Information about climate–growth relationships is crucial for predicting the potential climatic impact on tree species, especially those growing on the edges of their distribution range, for instance, in high-elevation forests. This study aimed to determine changes in the relationships between tree-ring widths and daily climatic data in high-elevation forests in the Western Carpathians over time. Climate–growth relationships were calculated to obtain the TRWI (tree-ring-width index) chronology (based on 104 trees) and day-wise aggregated data for temperature (mean, minimum, and maximum) and sums of precipitation. The radial growth of stone pine was mostly determined by the mean temperature in the period between mid-June (21st) and the beginning of July (4th) for the critical 14-day window width (r = 0.44). The negative influence of precipitation on the radial growth occurred in summer (r = −0.35) and overlapped with the period of the positive influence of temperature. Dendroclimatic studies based on daily data may define the exact periods (expressed in calendar days) that influence the radial growth of trees much better than the commonly used monthly means. This is particularly important in analysing the growth of trees at high elevations, where the climatic factor strongly limits radial growth. [ABSTRACT FROM AUTHOR]

Abstract Scatter-hoarding birds provide effective long-distance seed dispersal for plants. Transporting seeds far promotes population spread, colonization of new areas, and connectivity between populations. However, whether seeds transported over long distances are deposited in habitats favorable to plant regeneration has rarely been investigated, mainly due to methodological constraints. To investigate dispersal patterns and distances of Swiss stone pine (Pinus cembra) seeds we utilized advances in tracking technology to track the movements of their sole disperser, the spotted nutcracker (Nucifraga caryocatactes). We found routine individual movements between single seed harvesting and seed caching site. Harvesting sites of individual birds overlapped, whereas seed caching sites were separated and located on average 5.3 km away from the harvesting site. Interestingly, most distant caching sites were located at low elevations and in spruce forest, where Swiss stone pine does not naturally occur. This suggests that nutcrackers disperse seeds over long distances but that a large portion of these seeds are cached outside the known pine habitat. Therefore, we conclude that the implications of such long-distance seed dispersal movements for plant populations should be carefully considered in combination with the effects of habitat quality on plant recruitment.

Tree rings are an ecological indicator, useful for studying the adaptation of trees in a changing environment, especially in the unique habitat of high-elevation cliff forests. The main aim of this study was to characterize tree-ring changes in Stone pine (Pinus cembra L.) growing in the extreme conditions of cliff forests of the Western Carpathians (at elevation 1300–1600 m a.s.l.), in order to reconstruct the growth release pattern and to better understand the influence of climate. The study was based on 104 tree ring series covering a 449-year-long chronology (1561–2009). In the last century, mean annual temperature has increased by +2.0 °C and precipitation has decreased slightly in the Tatras. The growth of stone pine was strongly related to summer temperature of June and July. In the last decades, the negative influence of summer precipitation on tree growth has abated; this may be related to warmer summers and decreasing precipitation. Temperature of the previous autumn, winter, and early spring, which had a significant influence on stone pine growth in the past, is not an important factor now. In the extreme environment of mountain cliffs, the influence of temperature and precipitation on tree growth is not stable over time under changing climate. The current changes in climate seem to promote stone pine and its potential upward elevational shift. The numerous past disturbance events have affected a limited number of trees, suggesting that single individuals have been randomly eliminated from the cliff stands during the last three centuries. Our results highlight the complex pattern of the reaction of stone pine on cliffs to changes in the environment. Swiss stone pine, a long-lived tree species, shows high adaptability for colonization of mountain cliffs and is an excellent object for studying the response of plants to changing climate. Understanding the effect of interactions between climate, disturbances, and the radial growth of trees in extreme environments of cliff forests on different temporal scales (monthly, seasonal, yearly) should increase our knowledge of such ecologically valuable areas.

The chemical content of essential oils (EO) obtained from the leaves of four Pinaceae (Abies alba, Picea abies, Pinus cembra and Pinus mugo) was investigated by SPME-GC-MS technique. The vapor phase was characterized by the monoterpenes with values higher than 95.0%. Among them, α-pinene (24.7–48.5%), limonene (17.2–33.1%) and β-myrcene (9.2–27.8%) were the most abundant. The monoterpenic fraction prevailed over the sesquiterpenic one (≥74.7%) in the EO liquid phase. Limonene was the major compound in A. alba (30.4%), P. abies (20.3%) and P. mugo (78.5%), while α-pinene in P. cembra (36.2%). Regarding the phytotoxic properties, EOs were studied at different doses (2–100 μL) and concentrations (2–20/100 μL/mL). All EOs were found to be significantly active (p-value ˂ 0.05) against the two recipient species in a dose-dependent way. In pre-emergence tests, germination of Lolium multiflorum and Sinapis alba was reduced by up to 62–66% and 65–82%, respectively, as well as their growth by up to 60–74% and 65–67%, due to the effects of compounds in both the vapor and liquid phases. In post-emergence conditions, at the highest concentration, the phytotoxicity of EOs caused heavy symptoms and, in the case of S. alba, A. alba EO completely destroyed (100%) the treated seedlings. [ABSTRACT FROM AUTHOR]

Alpine treeline ecosystems are generally expected to advance with increasing temperatures and after land-use abandonment. Multiple interacting factors modify this trend. Understanding the long-term processes underlying treeline advance is essential to predict future changes in structure and function of mountain ecosystems. In a valley in the Central Swiss Alps, we re-assessed a 40-year-old survey of all treeline trees (>0.5 m height) and disentangled climate, topographical, biotic, and disturbance (land use and avalanche risk) factors that have led to treeline advance with a combination of ground-based mapping, decision tree, and dendroecological analyses. Between the first ground survey in 1972/73 and the resurvey in 2012, treeline advanced on average by 10 meters per decade with a maximum local advance of 42 meters per decade. Larch consistently advanced more on south-facing slopes, while pine advance was greater on north-facing slopes. Newly established spruce mostly represented infilling below the previous treeline. The forefront of treeline advance above 2330 m a.s.l. occurred mainly on favorable microsites without competing dwarf shrub vegetation. At slightly lower elevations, treeline advanced mainly on sites that were used for agriculture at the beginning of the 20th century. This study indicates that although treeline advances under the effect of climate warming, a combination of additional ecological factors controls this advance at regional and local scales. [ABSTRACT FROM AUTHOR]

The heartwood of the Swiss Stone Pine, Pinus cembra L., has been scarcely investigated for secondary metabolites for a long period of time. Considering age and relative simplicity of heartwood investigations dating back to the 1940s to 1960s, we conducted the first investigation of P. cembra heartwood by HPLC, using UHPLC-DAD-ESI-MSn and HPLC-DAD techniques in combination with isolation and NMR spectroscopy, with focus on stilbenes, bibenzyls and flavonoids. Analytical problems in the HPLC analysis of Pinus stilbenes and flavonoids on reversed stationary phases were also challenged, by comparing HPLC on pentafluorophenyl (PFP) and C18 stationary phases. Seven flavonoids (1, 2, 3, 7, 8, 11, 12), four stilbenes (4, 6, 10, 13), two bibenzyls (5, 9), three fatty acids (14, 16, 17) and one diterpenic acid (15) were detected in an ethanolic extract of Pinus cembra heartwood. HPLC comparison of reversed stationary phases in HPLC showed that the antifungal, antibacterial and chemosensitizing dihydropinosylvin monomethyl ether (9) and pinosylvin monomethyl ether (10) can be separated on PFP, but not on C18 material, when eluting with a screening gradient of 20–100% acetonitrile. Flavonoid separation showed additional benefits of combining analyses on different stationary phases, as flavonoids 7 and 8 could only be separated on one of two C18 stationary phases. Earlier phytochemical results for heartwood investigations were shown to be mostly correct, yet expandable. Substances 5 to 12 were found in alignment with these references, proving remarkable phytochemical analyses at the time. Evidence for the described presence of pinobanksin could not be found. Substances 1 to 4 and 13 have to our knowledge not yet been described for P. cembra.

Scatter-hoarding birds provide effective long-distance seed dispersal for plants. Transporting seeds far promotes population spread, colonization of new areas, and connectivity between populations. However, whether seeds transported over long distances are deposited in habitats favorable to plant regeneration has rarely been investigated, mainly due to methodological constraints. To investigate dispersal patterns and distances of Swiss stone pine (Pinus cembra) seeds we utilized advances in tracking technology to track the movements of their sole disperser, the spotted nutcracker (Nucifraga caryocatactes). We found routine individual movements between single seed harvesting and seed caching site. Harvesting sites of individual birds overlapped, whereas seed caching sites were separated and located on average 5.3 km away from the harvesting site. Interestingly, most distant caching sites were located at low elevations and in spruce forest, where Swiss stone pine does not naturally occur. This suggests that nutcrackers disperse seeds over long distances but that a large portion of these seeds are cached outside the known pine habitat. Therefore, we conclude that the implications of such long-distance seed dispersal movements for plant populations should be carefully considered in combination with the effects of habitat quality on plant recruitment. [ABSTRACT FROM AUTHOR]

pH of xylem sap (pHx) was determined in three trees (Malus domestica (apple tree), Picea abies and Pinus cembra) in response to seasonal changes. Conifer trees from lowland (600 m) were compared to trees growing at the alpine timberline (1950 m a.s.l.). Xylem sap was extracted with a Scholander pressure bomb and pHx was measured with a pH microsensor. In all species, pHx changed markedly with season. In spring, pHx was acidic; during winter, the pHx was more alkaline. In apple trees, the pHx did not show a significant correlation with temperature but was rather affected by developmental stage. During flushing in spring, xylem sap acidification took place concomitant to the developmental stage "tight cluster", when foliar development enables a significant transpiration and a consequent movement of water in the xylem. The xylem sap of the two studied conifers showed a significantly larger seasonal alkalinisation (+2.1) than found in apple trees (+1.2) and was significantly more pronounced at the timberline. Xylem sap acidification took place before bud break. pHx had a significant negative correlation with soil temperatures and corresponded to already reported pHx of angiosperms. Overall, pHx appears to be a sensitive stress marker and indicator of activity status in tree xylem. [ABSTRACT FROM AUTHOR]

The term 'Little Ice Age' (LIA) is classically used to define a period of repeated and extensive glacier advances during the last millennium. In the meanwhile, this term is also used to address the period of relatively low temperatures between the Medieval Climate Anomaly (MCA), or Medieval Warm Period, and present-day warming. The end of the LIA is generally set to the mid or late 1800s CE, however, the published onset dates of the LIA are more variable from the mid 1200s to the late 1500s. At Mont Miné and Morteratsch glaciers, Swiss Alps, we sampled and subsequently analysed detrital as well as in situ tree remnants from the early LIA period. At both glaciers, trees with lifespans of up to about 400 years were buried at various lateral moraine sites. The corresponding advance of both glaciers can be traced from the 1280s until the 1310s. At Morteratsch glacier, this early LIA advance phase culminated likely around 1375 CE. Evidence collected at both glaciers indicates that the ice surfaces were at least c. 12–15 m from the lateral moraine crests deposited during the maximum extent of the LIA. This suggests a similar (though very slightly weaker) magnitude than later LIA advances at our sites. The advances of Mont Miné and Morteratsch glaciers coincide with relatively cool summer temperatures from the late 1200s to the late 1300s. Taken together, the onset of the Little Ice Age in the Alps can be considered to be c. 1260 CE. The Little Ice Age was not a uniform period, but had several phases as can be derived from the records of Alpine glaciers and summer temperatures. We propose a subdivision of the LIA in the European Alps into an early (1260–1380 CE), an intermediate (1380–1575 CE) and a main (1575–1860 CE) phase. [ABSTRACT FROM AUTHOR]

The chemical content of essential oils (EO) obtained from the leaves of four Pinaceae (Abies alba, Picea abies, Pinus cembra and Pinus mugo) was investigated by SPME-GC-MS technique. The vapor phase was characterized by the monoterpenes with values higher than 95.0%. Among them, α-pinene (24.7–48.5%), limonene (17.2–33.1%) and β-myrcene (9.2–27.8%) were the most abundant. The monoterpenic fraction prevailed over the sesquiterpenic one (≥74.7%) in the EO liquid phase. Limonene was the major compound in A. alba (30.4%), P. abies (20.3%) and P. mugo (78.5%), while α-pinene in P. cembra (36.2%). Regarding the phytotoxic properties, EOs were studied at different doses (2–100 μL) and concentrations (2–20/100 μL/mL). All EOs were found to be significantly active (p-value ˂ 0.05) against the two recipient species in a dose-dependent way. In pre-emergence tests, germination of Lolium multiflorum and Sinapis alba was reduced by up to 62–66% and 65–82%, respectively, as well as their growth by up to 60–74% and 65–67%, due to the effects of compounds in both the vapor and liquid phases. In post-emergence conditions, at the highest concentration, the phytotoxicity of EOs caused heavy symptoms and, in the case of S. alba, A. alba EO completely destroyed (100%) the treated seedlings.

Due to anthropogenic impact on vegetation cover, importance of the outphytosozological studies of coniferous plants of the Ukrainian Carpathians is emphasized. Pinus cembra is one of the rare species of trees in Europe and its in situ and ex situ conservation in Ukraine is required. At present, P. cembra is under protection in its natural habitats within 25 objects of the nature reserve fund of Ukraine and 71 objects of artificial plantations (in total of 10 categories). The network of the protected areas consists of two biosphere reserves, one nature reserve, six national nature parks, one regional landscape park, eight reserves (of which five are botanical), 13 botanical nature monuments, seven protected forest parcels, 14 botanical gardens, 16 dendrological parks, and 21 monuments of landscape art. The largest natural plant communities of P. cembra are located in Gorgany Nature Reserve (539 ha). As a result of the inventory studies, the boundaries of its cultivation area in Ukraine were defined. It is obvious that the most widespread cultivation of P. cembra is within the Carpathian Mountains (20 objects). The Forest-Steppe zone of Ukraine and the zone of deciduous forests of Ukraine (14 and 16 objects, respectively) provide sufficiently suitable ecological conditions for cultivation of P. cembra. The least number of artificial plantations under protection were found in the Mixed Forest zone (4) and the Steppe zone of Ukraine (6 objects). The largest number of the protected natural forests (16) and artificial plantations (11 objects) was registered in Ivano-Frankivsk Region. Plants aged ca. 250 years were revealed under artificial conditions (in cultivation).

Abstract Background Genetic processes shape the modern-day distribution of genetic variation within and between populations and can provide important insights into the underlying mechanisms of evolution. The resulting genetic variation is often unequally partitioned within species’ distribution range and especially large differences can manifest at the range limit, where population fragmentation and isolation play a crucial role in species survival. Despite several molecular studies investigating the genetic diversity and differentiation of European Alpine mountain forests, the climatic and demographic constrains which influence the genetic processes are often unknown. Here, we apply non-coding microsatellite markers to evaluate the sporadic peripheral and continuous populations of cembra pine (Pinus cembra L.), a long-lived conifer species that inhabits the subalpine treeline ecotone in the western Alps to investigate how the genetic processes contribute to the modern-day spatial distribution. Moreover, we corroborate our findings with paleoecological records, micro and macro-remains, to infer the species’ possible glacial refugia and expansion scenarios. Results Four genetically distinct groups were identified, with Bayesian and F ST based approaches, across the range of the species, situated in the northern, inner and south-western Alps. We found that genetic differentiation is substantially higher in marginal populations than at the center of the range, and marginal stands are characterized by geographic and genetic isolation due to spatial segregation and restricted gene flow. Moreover, multiple matrix regression approaches revealed effects of climatic heterogeneity in species’ spatial genetic pattern. Also, population stability tests indicated that all populations had experienced a severe historical bottleneck, no heterozygosity excess was detected, suggesting that more recently population sizes have remained relatively stable. Conclusions Our study demonstrated that cembra pine might have survived in multiple glacial refugia and subsequently recolonized the Alps by different routes. Modern-day marginal populations, at the edge of the species’ range, could maintain stable sizes over long periods without inbreeding depression and preserve high amounts of genetic variation. Moreover, our analyses indicate that climatic variability has played a major role in shaping differentiation, in addition to past historical events such as migration and demographic changes.

Pinus cembra L. is a coniferous European mountain range tree rich in oleoresins and essential oils. Twig tips with needles are the most common tree parts used to obtain essential oils. As the whole tree contains volatiles, the essential oil composition from different parts was studied, including twig tips with needles from the bottom, the top and the cone bearing branches, cones, twigs without needles from the lower and upper part of the crown as well as wood and bark from the trunk. The variability in essential oil composition between these plant parts and between individual trees has been studied using multivariate statistical analyses. α-Pinene was present in all samples, being highest in cones (49.3%) and lowest in sapwood (0.7%). Twig tips with needles from different parts of the crown had similar essential oil composition with α-pinene (43.9–48.3%), β-phellandrene (13.1–17.2%), β-pinene (6.6–9.3%), germacrene D (5.1–6.8%) and limonene (4.1–6.1%) as main compounds. Twig essential oils had usually more β-phellandrene (23.9–29.8%) than α-pinene (23.3–24.3%) and also appreciable amounts of β-pinene (13.5–15.1%) and limonene (11.9–17.9%). Cone essential oils contained mainly α-pinene, β-pinene (20.1%) and limonene (13.9%). The essential oils from wood and bark were rich in diterpenes as cembratrienol (4.9–21.4%), cembrene (4.8–14.3%) and methyl daniellate (2.7–6.8%). Sapwood distillates contained also notable amounts of alkane derivatives. Finally, the solvent free SPME technique has been employed to analyse the volatiles in the plant parts. For needles and cones the observed patterns were in good accordance with the compositions of the respective essential oils. [ABSTRACT FROM AUTHOR]

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Although several studies provide a broad overview of vegetation changes in the Carpathian Basin during the Holocene, stand-scale vegetation changes are lesser known because of the rarity of suitable sampling sites. In this study we investigated the sediment of a small closed-canopy site (Nagy-forrás forest hollow, 685 m a.s.l., 0.1 ha), located in the Mátra Mountains, on the north facing slope of Kékes (1014 m a.s.l.). We carried out detailed pollen, conifer stomata and plant macrofossil analyses, as well as radiocarbon dating to examine Late Glacial and Holocene dynamics of vegetation development. The site dates back to ca. 15,500 cal yr BP, when open boreal forests and wet tundra-like habitats occurred around the hollow. Closed forest cover developed around 14,600 cal yr BP, when a boreal European larch-Swiss stone pine (Larix decidua-Pinus cembra) forest surrounded the hollow. This vegetation type remained stable up to 7700 cal yr BP. We observed a hiatus between 7700 and 2710 cal yr BP, followed by a beech (Fagus sylvatica) dominated mixed temperate deciduous forest. Our results confirmed that the area was covered by a primary forest, as human influence was visible only from 175 cal yr BP. The relatively long lasting persistence of Pinus cembra in the Holocene at relatively low altitude was documented, which has never been found in Holocene sediments in the Pre-Carpathians before. We hypothesize that the north facing slope acted as a cold-stage refugium in the Early Holocene and could play the same role for the present-day beech forest that is threatened by recent climate change. [ABSTRACT FROM AUTHOR]

Widespread and locally severe foliar symptoms resembling Dothistroma Needle Blight (DNB), one of the most important infectious diseases of forest trees worldwide, were recently observed in La Sila Massif, a mountain plateau covered with native forests of Pinus nigra subsp. laricio in La Sila National Park, Southern Italy. At the same time, DNB symptoms were observed in Pinus cembra and Pinus mugo forests in the Paneveggio-Pale di San Martino Nature Park and in Val Sarentino, Northeastern Italy. Defoliation was extensive at all sites and severe on the majority of plants of affected species, both adult trees and renovation. In particular on Pinus cembra, the disease was so serious as to locally threaten the species' reproduction and survival, an unusually heavy damage on this host. Species-specific real time PCR diagnostics, recommended by EPPO, was applied to needle samples from these sites and the presence of Dothistroma septosporum was ascertained, while Dothistroma pini, the morphologically identical congeneric species causing the same disease, was not detected. The pathogen was isolated from conidiomata and species attribution was confirmed by sequencing of the ITS region. In addition, a culture independent survey based on the same molecular assays was carried out in other areas of Northern, Central and Southern Italy, where pines of various species were affected by similar symptoms, and gave negative results for both Dothistroma species. Results show that D. septosporum currently has in Italy a much larger distribution and host range than reported and is associated to life-threatening damage to native pine species growing in established populations for in situ conservation of genetic resources, which would require an update of specific conservation actions. [ABSTRACT FROM AUTHOR]

Key message: The lack of elevational changes in electrical resistivity in Pinus cembra trunks indicated consistent growth and hydraulics across elevations. Though, electrical resistivity tomograms exhibited pronounced temperature-driven seasonal changes. Alpine conifers growing at high elevation are exposed to low temperatures, which may limit xylogenesis and cause pronounced seasonal changes in tree hydraulics. Electrical resistivity (ER) tomography enables minimal invasive monitoring of stems in situ. We used this technique to analyze Pinus cembra trunks along a 400 m elevational gradient up to the timberline and over seasons. Furthermore, ER data of earlywood across tree rings were compared with the respective specific hydraulic conductivity (KS), measured on extracted wood cores. ER tomograms revealed pronounced changes over the year and a strong correlation between average ER (ERmean) and air and xylem temperatures. Surprisingly, no elevational changes in ERmean, earlywood ER or KS were observed. ER data corresponded to variation in earlywood KS, which decreased from the youngest (ca. 4–5 cm2s−1 MPa−1) to the oldest tree rings (0.63 ± 0.22 cm2s−1 MPa−1). The lack of changes in ER data and earlywood KS along the study transect indicated consistent growth patterns and no major changes in structural and functional hydraulic traits across elevation. The constant decrease in earlywood KS with tree ring age throughout all elevations highlights the hydraulic relevance of the outermost tree rings in P. cembra. Seasonal measurements demonstrated pronounced temperature effects on ER, and we thus recommend a detailed monitoring of trunk temperatures for ER tomography. [ABSTRACT FROM AUTHOR]

It is generally accepted that the spatial distribution of neutral genetic diversity within a species' native range mostly depends on effective population size, demographic history, and geographic position. However, it is unclear how genetic diversity at adaptive loci correlates with geographic peripherality or with habitat suitability within the ecological niche. Using exome‐wide genomic data and distribution maps of the Alpine range, we first tested whether geographic peripherality correlates with four measures of population genetic diversity at > 17,000 SNP loci in 24 Alpine populations (480 individuals) of Swiss stone pine (Pinus cembra) from Switzerland. To distinguish between neutral and adaptive SNP sets, we used four approaches (two gene diversity estimates, FST outlier test, and environmental association analysis) that search for signatures of selection. Second, we established ecological niche models for P. cembra in the study range and investigated how habitat suitability correlates with genetic diversity at neutral and adaptive loci. All estimates of neutral genetic diversity decreased with geographic peripherality, but were uncorrelated with habitat suitability. However, heterozygosity (He) at adaptive loci based on Tajima's D declined significantly with increasingly suitable conditions. No other diversity estimates at adaptive loci were correlated with habitat suitability. Our findings suggest that populations at the edge of a species' geographic distribution harbour limited neutral genetic diversity due to demographic properties. Moreover, we argue that populations from suitable habitats went through strong selection processes, are thus well adapted to local conditions, and therefore exhibit reduced genetic diversity at adaptive loci compared to populations at niche margins. [ABSTRACT FROM AUTHOR]

Massively parallel sequencing of cDNA is an efficient route for generating sequence collections that represent expressed genes under different environmental control. The analysis of their sequence helps in developing molecular markers, such as SNPs, which represent a useful tool in detecting adaptive signals in populations. In this study novel PCR markers, based on stress responsive genes, were designed from the transcriptome of the haploxylon Swiss stone pine (Pinus cembra L.) and tested for SNPs in the diploxylon Scots pine (Pinus sylvestris L.). 84 primers were tested on P. sylvestris DNA samples originating from three different types of habitat. After sequencing and BLAST search of the amplified products, parts of 19 different candidate genes were analysed by considering the polymorphic sites, insertions/deletions as well as synonymous and non-synonymous SNPs. In a total of 3735 sites no indels, eight synonymous and 11 non-synonymous SNPs were found. By providing de novo molecular markers developed in P. cembra and tested for transferability in Scots pine, our results give support for the use of de novo markers targeting conserved regions across different pines. The SNPs detected may have important applications in further studies of adaptive genetic variation, providing tools to study relevant genes important in the long-term adaptation of pine species. [ABSTRACT FROM AUTHOR]

Key Message: Mosaicism has been confirmed in several studied old trees from both gymnosperms and angiosperms, in a pattern not always following the actual branching of the tree. Eight individuals from two gymnosperm and three angiosperm species of trees were studied for age-related mosaicism. We compare the level of intra-organismal genetic variability found within an individual old and an individual young tree of the same species to prove its relation to age. In five cases, we found statistically significant differences in intra-organismal genetic variability between young and old trees. In three cases of mosaicism, the constructed dendrogram corresponded with the actual branching of the tree, but it differed in the remaining two. We offer an explanation of the cases when the dendrogram does not correspond with the branching structure of the tree through the patterns of sectorial and mericlinal chimerism. In the studied gymnosperms, we found that the growth rate possibly influenced the level of intra-organismal variability, while in angiosperms the dependence was rather on age. The differences between angiosperm and gymnosperm molecular evolution and meristem stratification may play a role in this, but further research is needed in this area. [ABSTRACT FROM AUTHOR]

Key message: The axial structure of the hydraulic system in trees is relatively invariant and insensitive to temperature, while trees plastically adjust the number of cells within the tree ring. At higher elevations and latitudes in the treeline ecotone, reduction in the heat accrued during the growing season is reflected in gradually decreasing tree size. Due to low temperatures, treeline trees might produce smaller xylem cells and, as a consequence, tree growth could be limited. However, some xylem traits (i.e., cell lumen area) are considered relatively insensitive to climatic factors but highly dependent on tree size because of the natural widening of xylem conduits towards the stem base. We tested the hypothesis that earlywood cell lumen area is essentially invariant and depends largely on tree size. Tracheid traits in four conifer species from the lower ("timberline") and upper ("treeline") parts of the treeline ecotone (Picea engelmannii, Picea abies, Pinus cembra and Larix decidua) were measured in the Colorado Front Range (U.S.A.), Krkonoše Mts. (Czech Republic) and Dolomites (Italy). On transversal sections sampled at 1 m of stem height, we measured cell lumen areas, transversal cell size, cell wall thickness, tree-ring width and number of cells per radial file. Cell lumen areas were always greater at the timberline than treeline. When tree height is accounted for, the earlywood cell area did not differ between the two sites, thus showing that difference in temperature did not affect earlywood cell area in any of the four measured species. The number of cells within tree rings exhibited high inter-annual variability according to environmental factors. The fundamental hydraulic structure in trees is relatively rigid and insensitive to temperature, while trees plastically adjust the number of cells within the tree ring as a result of inter-annual climate variability and leaf production. [ABSTRACT FROM AUTHOR]

Beside low temperatures, limited tree growth at the alpine treeline may also be attributed to a lack of available soil nutrients and competition with understory vegetation. Although intra-annual stem growth of Pinus cembra has been studied intensively at the alpine treeline, the responses of radial growth to soil warming, soil fertilization, and below ground competition awaits clarification. In this study we quantified the effects of nitrogen (N) fertilization, soil warming, and understory removal on stem radial growth of P. cembra at treeline. Soil warming was achieved by roofing the forest floor with a transparent polyvinyl skin, while understory competition was prevented by shading the forest floor with a non-transparent foil around six trees each. Six trees received N- fertilization and six other trees served as controls. Stem growth was monitored with band dendrometers during the growing seasons 2012–2014. Our 3 years experiment showed that soil warming had no considerable effect on radial growth. Though understory removal through shading was accompanied by root-zone cooling, understory removal as well as N fertilization led to a significant increase in radial growth. Hardly affected was tree root biomass, while N-fertilization and understory removal significantly increased in 100-needle surface area and 100-needle dry mass, implying a higher amount of N stored in needles. Overall, our results demonstrate that beside low temperatures, tree growth at cold-climate boundaries may also be limited by root competition for nutrients between trees and understory vegetation. We conclude that tree understory interactions may also control treeline dynamics in a future changing environment.

Climate sensitivity of populations at the margins of their distribution range is of key importance to understand species’ responses to future warming conditions. Pinus cembra is of particular interest being a typical high-elevation taxon, spread with mostly scattered populations within its actual range, but still overlooked in traditional dendrochronological researches due to low tree-ring variability and climate sensitivity. With a different approach, we analyzed time series of xylem anatomical traits, split into intra-ring sectors, and used daily climate records over 89 years (1926-2014) aiming to improve the quality and time resolution of the climate/growth associations. From nine trees growing at their altitudinal limit and on 1.5 × 106 tracheids, we measured ring width (MRW), cell number per ring, lumen area (LA), and cell-wall thickness (CWT). We then computed correlations with monthly and fortnightly climate data. Late-spring and summer temperature emerged as the most important factors. LA and especially CWT showed a stronger temperature response than MRW, starting in mid-May and early June, respectively. CWT also evidenced the longest period of correlations with temperature and a significant difference between latewood radial and tangential walls. Analysis of xylem anatomical traits at intra-ring level and the use of daily temperature records proved to be useful for high resolution and detailed climate/growth association inferences in Pinus cembra. [ABSTRACT FROM AUTHOR]

Abstract: Fungi are both agents of disease and mutualistic partners of plants. Previous studies have tested the effects of abiotic or biotic factors on plant‐associated fungal communities in isolation. However, to better understand patterns of plant–fungal associations, the combined effects of abiotic and biotic drivers across environmental gradients may be important. We investigated the effects of temperature, pH, soil moisture, vegetation cover and distance to host plant on the occurrence and abundance of fungi associated with Swiss stone pine (Pinus cembra). We did this by DNA metabarcoding 288 soil samples taken across and beyond the elevation range of P. cembra (i.e. 1,850–2,250 m a.s.l.) in two valleys in the Swiss Alps. We modelled the effects of abiotic and biotic factors on DNA read abundance of pathogenic and mutualistic fungal operational taxonomic units (OTUs) associated with P. cembra. We also tested whether abiotic and biotic factors differentially affected fungi of varying host specificity (i.e. host generalists, host specialists). We found that the occurrences of both host generalist and specialist fungi exceeded the current elevational range of their host plant. Abiotic factors had only minor effects on the abundances of all fungal OTUs. However, we found positive effects of the host plant on the abundance of a host specialist pathogenic fungus, providing support for a Janzen–Connell effect of high pathogen accumulation close to conspecific host plants. We also found a positive response to the host plant in a specialist ectomycorrhizal fungus, suggesting an “inverse” Janzen–Connell effect. Synthesis. Our findings imply that negative distance dependence shapes not only the distribution of host‐specific fungal pathogens, but also host‐specific fungal mutualists. We conclude that the occurrence of both pathogenic and mutualistic fungi beyond the current elevational range of host plants may determine their potential range shifts under projected climate warming. [ABSTRACT FROM AUTHOR]

Beside low temperatures, limited tree growth at the alpine treeline may also be attributed to a lack of available soil nutrients and competition with understory vegetation. Although intra-annual stem growth of Pinus cembra has been studied intensively at the alpine treeline, the responses of radial growth to soil warming, soil fertilization, and below ground competition awaits clarification. In this study we quantified the effects of nitrogen (N) fertilization, soil warming, and understory removal on stem radial growth of P. cembra at treeline. Soil warming was achieved by roofing the forest floor with a transparent polyvinyl skin, while understory competition was prevented by shading the forest floor with a non-transparent foil around six trees each. Six trees received N-fertilization and six other trees served as controls. Stem growth was monitored with band dendrometers during the growing seasons 2012-2014. Our 3 years experiment showed that soil warming had no considerable effect on radial growth. Though understory removal through shading was accompanied by root-zone cooling, understory removal as well as N fertilization led to a significant increase in radial growth. Hardly affected was tree root biomass, while N-fertilization and understory removal significantly increased in 100-needle surface area and 100-needle dry mass, implying a higher amount of N stored in needles. Overall, our results demonstrate that beside low temperatures, tree growth at cold-climate boundaries may also be limited by root competition for nutrients between trees and understory vegetation. We conclude that tree understory interactions may also control treeline dynamics in a future changing environment. [ABSTRACT FROM AUTHOR]

In alpine regions, tree hydraulics are limited by low temperatures that restrict xylem growth and induce winter frost drought and freezing stress. While several studies have dealt with functional limitations, data on elevational changes in functionally relevant xylem anatomical parameters are still scarce. In wood cores of Pinus cembra L. and Picea abies (L.) Karst. trunks, harvested along five elevational transects, xylem anatomical parameters (tracheid hydraulic diameter dh, wall reinforcement (t/b)2), pit dimensions (pit aperture Da, pit membrane Dm and torus Dt diameters) and respective functional indices (torus overlap O, margo flexibility) were measured. In both species, tracheid diameters decreased and (t/b)² increased with increasing elevation, while pit dimensions and functional indices remained rather constant (P. cembra: Dt 10.3 ± 0.2 μm, O 0.477 ± 0.005; P. abies: Dt 9.30 ± 0.18 μm, O 0.492 ± 0.005). However, dh increased with tree height following a power trajectory with an exponent of 0.21, and also pit dimensions increased with tree height (exponents: Dm 0.18; Dt 0.14; Da 0.11). Observed elevational trends in xylem structures were predominantly determined by changes in tree size. Tree height-related changes in anatomical traits showed a remarkable robustness, regardless of the distributional ranges of study species. Despite increasing stress intensities towards the timberline, no adjustment in hydraulic safety at the pit level was observed. [ABSTRACT FROM AUTHOR]

The analysis of the anatomical characteristics of tree rings of P. sylvestris L. rootstocks and scions of P. sibirica Du Tour (Emelianovo and Surgut provenances) and P. cembra L. (Karpat provenance) has been carried out for 10 years of their joint growth. The number of tracheids, their radial diameter and the cell wall thickness for the P. sibirica scions increases relative to control trees (with the exception of the cell wall thickness for Surgut scions). At the same time, in the annual rings of the P. sylvestris rootstocks these characteristics as well as the number of tracheids decrease relative to the control trees. An exception is the Scots pine, which is the rootstock for Surgut provenance scion, for which the radial tracheid diameter of the transition zone increases. The seasonal dynamics of the radial tracheid diameter and the cell wall thickness of the grafted trees retains the species features typical for the ungrafted trees, suggesting the stability of the genetic program of xylem differentiation. The adaptation of scions to the climatic conditions of their place of origin does not affect the anatomical structure of the tree rings of the rootstocks during the years in which the conditions are close to the climate of the region of origin of the scions

Treeline position is mainly determined by growth season temperature, but the response of treelines to climate warming is not uniform worldwide. We compared treeline structure, dynamics and thermal profile in nearby areas with different treeline type, species composition and bioclimatic conditions. We performed a detailed survey of different treeline types in three areas of Italian Alps and northern Apennines. Every tree individual was recorded along altitudinal transects from the closed forest to the species limit. Treeline structure and dynamics were described through altitudinal limits of tree height, density and age. Data were elaborated by principal components analysis. Temperature regime of the three sites was assessed from homogeneous historical climatic data. Treeline was different in the three areas for species composition, shape and dynamics. Both Alpine sites showed diffuse treeline, but only one showed advancing dynamics. Apennine treeline was abrupt and static, with higher temperature at the tree limit. Our study showed the variable dynamics of treelines within a relatively restricted area and the connection between treeline shape and dynamics. An important role is played by species composition, determined by bioclimatic and historical features. These factors should be taken into account when modelling future treeline dynamics at global scale. [ABSTRACT FROM AUTHOR]

European larch ( Larix decidua) forests of the western Alps form extensive cultural landscapes whose resilience to global changes is currently unknown. Resilience describes the capacity of ecological systems to maintain the same state, i.e., the same function, processes, structure, and composition despite disturbances, environmental changes and internal fluctuations. Our aim is to explore the resilience of larch forests to changes in climate and land use in the western Italian Alps. To do so, we examined whether larch forests can be described as an alternative stable state in mountain forest ecosystems. We used tree basal area data obtained from field forest inventories in combination with topography, forest structure, land use, and climate information. We applied three different probabilistic methods: frequency distributions, logistic regressions, and potential analyses to infer the resilience of larch forests relative to that of other forest types. We found patters indicative of alternative stable states: bimodality in the frequency distribution of the percent of larch basal area, and the presence of an unstable state, i.e., mixed larch forests, in the potential analyses. We also found: (1) high frequency of pure larch forests at high elevation, (2) the probability of pure larch forests increased mostly with elevation, and (3) pure larch forests were a stable state in the upper montane and subalpine belts. Our study shows that the resilience of larch forests may increase with elevation, most likely due to the altitudinal effect on climate. Under the same climate conditions, land use seems to be the main factor governing the dominance of larch forests. In fact, subalpine larch forests may be more resilient, and natural succession after land abandonment, e.g., towards Pinus cembra forests, seems slower than in montane larch forests. In contrast, in the upper montane belt only intense land use regimes characterized by open canopies and forest grazing may maintain larch forests. We conclude that similar approaches could be applied in other forest ecosystems to infer the resilience of tree species. [ABSTRACT FROM AUTHOR]

[Display omitted] • Summer temperature remained a key factor affecting the growth of Swiss stone pine. • Summer precipitation is not the main factor limiting the growth of Swiss stone pine now. • The climate-growth relationship in stone pine is not stable over time. • Small-scale disturbances on cliffs have randomly affected trees over the last centuries. • The disturbance regime cannot be linked to any specific climatic extremes in the Tatras. Tree rings are an ecological indicator, useful for studying the adaptation of trees in a changing environment, especially in the unique habitat of high-elevation cliff forests. The main aim of this study was to characterize tree-ring changes in Stone pine (Pinus cembra L.) growing in the extreme conditions of cliff forests of the Western Carpathians (at elevation 1300–1600 m a.s.l.), in order to reconstruct the growth release pattern and to better understand the influence of climate. The study was based on 104 tree ring series covering a 449-year-long chronology (1561–2009). In the last century, mean annual temperature has increased by +2.0 °C and precipitation has decreased slightly in the Tatras. The growth of stone pine was strongly related to summer temperature of June and July. In the last decades, the negative influence of summer precipitation on tree growth has abated; this may be related to warmer summers and decreasing precipitation. Temperature of the previous autumn, winter, and early spring, which had a significant influence on stone pine growth in the past, is not an important factor now. In the extreme environment of mountain cliffs, the influence of temperature and precipitation on tree growth is not stable over time under changing climate. The current changes in climate seem to promote stone pine and its potential upward elevational shift. The numerous past disturbance events have affected a limited number of trees, suggesting that single individuals have been randomly eliminated from the cliff stands during the last three centuries. Our results highlight the complex pattern of the reaction of stone pine on cliffs to changes in the environment. Swiss stone pine, a long-lived tree species, shows high adaptability for colonization of mountain cliffs and is an excellent object for studying the response of plants to changing climate. Understanding the effect of interactions between climate, disturbances, and the radial growth of trees in extreme environments of cliff forests on different temporal scales (monthly, seasonal, yearly) should increase our knowledge of such ecologically valuable areas. [ABSTRACT FROM AUTHOR]

Analysis of the radial growth and of the climatic response of heterografts of Siberian pine (Pinus sibirica Du Tour) and Swiss pine (Pinus cembra L.) from three provenances was conducted. In all of the cases the rootstock was Scots pine (Pinus sylvestris L.). Radial growth of the grafts of Siberian pine was 23 % higher than for Swiss pine due to species peculiarities. The radial increment of the rootstock of Scots pine depends on the type of scion: the diameter growth of P. sylvestris (rootstock for Siberian pine) is greater than for the control trees by 20 % for the Surgut provenance and by 15 % for the Emelianovo provenance, whereas the diameter growth of Scots pine (rootstock for Swiss pine) is 10 % less than for the control stand (ungrafted trees). This can be considered to be a result of the scion influence. Analysis of climatic response of scion and rootstock shows that their reaction to air temperature is identical to the reaction of the control trees (positive correlation with April temperature). However, when the component representing the climatic reaction of the control trees is subtracted out, other results are obtained: the climatic response is absent for the rootstocks, whereas the indexes of the growth of scion are positively correlated with the temperature of autumn period and negatively correlated with the temperature of April-May.