Allergenicity and structural properties of new Cor a 1 isoallergens from hazel identified in different plant tissues.

The hazel allergen Cor a 1 is a PR-10 protein, closely related to the major birch pollen allergen Bet v 1. Hazel allergies are caused by cross-reactive IgE antibodies originally directed against Bet v 1. Despite the importance of PR-10 proteins in allergy development, their function and localization in the plant remain largely elusive. Therefore, the presence of Cor a 1 mRNA and proteins was investigated in different tissues, i.e., the female flower, immature and mature nuts, catkins, and pollen. Four yet unknown Cor a 1 isoallergens, i.e., Cor a 1.0501-1.0801, and one new Cor a 1.03 variant were discovered and characterized. Depending on the isoallergen, the occurrence and level of mRNA expression varied in different tissues, suggesting different functions. Interestingly, Cor a 1.04 previously thought to be only present in nuts, was also detected in catkins and pollen. The corresponding Cor a 1 genes were expressed in Escherichia coli. The purified proteins were analysed by CD and NMR spectroscopy. Immunoblots and ELISAs to determine their allergenic potential showed that the new proteins reacted positively with sera from patients allergic to birch, hazel and elder pollen and were recognized as novel isoallergens/variants by the WHO/IUIS Allergen Nomenclature Sub-Committee.

Assessment of the phenotypic and genotypic diversity of endophytic strains of Bacillus and closely related genera from Carpinus betulus in the Hyrcanian forests of Iran.

BACKGROUND: Identification and characterization of the endophytic microorganism, is gaining their underestimated significance in influencing health, performance, and other biological attributions of plants in general and forest tree species in particular. Because of the scarcity of information on the endophytic microbiome of the Hyrcanian forests species, including hornbeam (Carpinus betulus L.) trees, as a major constituent thereof, the present study aimed at the identification and partial characterization of the endophytic Bacillus species of Carpinus betulus as the first step in this context. METHODS AND RESULTS: Shoot samples were collected from the Hyrcanian forest locations of Mazandaran and Golestan provinces in Iran. Bacterial strains were isolated from the surface-disinfected shoot segments and subjected to phenotypic characterization. Following assessment of the genetic diversity of the isolates by BOX-PCR fingerprinting, the representative isolates of each of the 15 groups were used for further characterization. Analysis of the nucleotide sequences of the 16S rDNA and HSP60 gene of the isolates led to the identification of 10 species. The predominant species was B. cereus followed by B. subtilis. The other species encountered were B. thuringiensis, Priestia filamentosa, B. velezensis, B. mojavensis, B. amyloliquefaciens, B. safensis, P. aryabhattai, and Gottfriedia acidiceleris. Most isolates possessed characteristics which could contribute to the biocontrol potential of the isolates, including formation of biofilm, production of hydrogen cyanide, tolerant to relatively high concentration of sodium chloride, and antibacterial activity. CONCLUSIONS: Ten Bacillus species were identified as the prevailing endophytic species of C. betulus in the Hyrcanian forest of northern Iran, most turned up to possess biological activities involved in biocontrol capability of the isolates against some plant pathogens. These potentially capable bacteria could be implemented in the promotion of plant growth as well as in the biological control of pathogens. This is the first report on the characterization and elucidation of the diversity of the potentially beneficial endophytic species of Bacillus and the closely related genera living in the internal tissues of hornbeam trees.

Effects of birch encroachment, water table and vegetation on methane emissions from peatland microforms in a rewetted bog.

This study investigated the influence of vegetation and microforms on methane (CH4) balances of a rewetted bog in north-west Germany. The two study sites are in close proximity on the same former peat extraction area, one dominated by Sphagnum-mosses and the other one by a dense Betula pubescens stand with a high Eriophorum vaginatum cover. The contribution of microforms (hummocks/hollows) to CH4 emissions and the effect of Betula encroachment has been studied. Transparent and opaque chambers were used to measure CH4 fluxes every 3-4 weeks during daytime for one year. For the estimation of annual balances, three methods were compared and the method using water level and soil temperature as explanatory variables was selected. Fluxes were scaled to the site level. The annual emissions per site are and 7.1 ± 1.5 g CH4-C m-2 year-1 at the treed site and 36.1 ± 3.5 g CH4-C m-2 year-1 at the open site, mainly controlled by higher water levels. Highest annual emissions originated from hollows at the open site, but in the vegetation period, hummock emissions tend to be higher. At the tree site, emission differences between the microforms were less pronounced. There were no differences between fluxes from transparent and opaque chambers.

Delayed postglacial colonization of Betula in Iceland and the circum North Atlantic.

As the Arctic continues to warm, woody shrubs are expected to expand northward. This process, known as 'shrubification,' has important implications for regional biodiversity, food web structure, and high-latitude temperature amplification. While the future rate of shrubification remains poorly constrained, past records of plant immigration to newly deglaciated landscapes in the Arctic may serve as useful analogs. We provide one new postglacial Holocene sedimentary ancient DNA (sedaDNA) record of vascular plants from Iceland and place a second Iceland postglacial sedaDNA record on an improved geochronology; both show Salicaceae present shortly after deglaciation, whereas Betulaceae first appears more than 1000 y later. We find a similar pattern of delayed Betulaceae colonization in eight previously published postglacial sedaDNA records from across the glaciated circum North Atlantic. In nearly all cases, we find that Salicaceae colonizes earlier than Betulaceae and that Betulaceae colonization is increasingly delayed for locations farther from glacial-age woody plant refugia. These trends in Salicaceae and Betulaceae colonization are consistent with the plant families' environmental tolerances, species diversity, reproductive strategies, seed sizes, and soil preferences. As these reconstructions capture the efficiency of postglacial vascular plant migration during a past period of high-latitude warming, a similarly slow response of some woody shrubs to current warming in glaciated regions, and possibly non-glaciated tundra, may delay Arctic shrubification and future changes in the structure of tundra ecosystems and temperature amplification.

Perhaps there were northern refugia in LGM? The phylogeographic structure of the thermophilic tree Carpinus betulus in the Carpathian region.

Carpinus betulus L., the hornbeam, is a component of lowland and highland forests in Europe. By examining the postglacial migratory history of thermophilic tree species, the study aimed to unravel their putative glacial microrefugia in the Carpathian region. The present study points to the two distinct genetic AFLP groups of C. betulus in the Carpathian region that represent different genetic lineages based on Bayesian analysis. They differed in Nei's gene diversity index h, and the analysis of molecular variance AMOVA showed a percentage variation of the populations between the groups of 13.74 %. Principal coordinate analysis (PCoA) of 368 AFLP tree samples confirmed the presence of two genetic groups. Ninety-five populations underwent principal component analysis (PCA) to show the main correlations between genetic diversity indices and bioclimatic/climate variables (WorldClim and Carpatclim). The generalized logistic model (GLM) showed the significance of Nei's genetic index h in delimiting genetic groups. The results of population-genetic and multivariate analyses determined that the two genetic groups nowadays are spatially diffused and do not show a clear geographic pattern, pointing to a genetic melting pot. We found ecological links between genetic diversity and bioclimatic characteristics, especially the precipitation in the coldest quarter - Bio19. The refugial Maxent model indicates a significant contribution of the Bio7 variable (both linked with a continental type of climate) to the occurrence of the species during the LGM in Europe. We suggest the relict character of hornbeam populations in a specific climatic-terrain niche in the northern part of the Carpathian Basin.

Evaluation of the Chemical Stability, Membrane Permeability and Antiproliferative Activity of Cyclic Diarylheptanoids from European Hornbeam (Carpinus betulus L.).

Four cyclic diarylheptanoids-carpinontriols A (1) and B (2), giffonin X (3) and 3,12,17-trihydroxytricyclo [12.3.1.12,6]nonadeca-1(18),2(19),3,5,14,16-hexaene-8,11-dione (4)-were isolated from Carpinus betulus (Betulaceae). Chemical stability of the isolated diarylheptanoids was evaluated as a function of storage temperature (-15, 5, 22 °C) and time (12 and 23 weeks). The effect of the solvent and the pH (1.2, 6.8, 7.4) on the stability of these diarylheptanoids was also investigated. Compounds 2 and 4 showed good stability both in aqueous and methanolic solutions at all investigated temperatures. Only 2 was stable at all three studied biorelevant pH values. Degradation products of 1 and 3 were formed by the elimination of a water molecule from the parent compounds, as confirmed by ultrahigh-performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-HR-MS). The permeability of the compounds across biological membranes was evaluated by the parallel artificial membrane permeability assay (PAMPA). Compound 3 possesses a logPe value of -5.92 ± 0.04 in the blood-brain barrier-specific PAMPA-BBB study, indicating that it may be able to cross the blood-brain barrier via passive diffusion. The in vitro antiproliferative activity of the compounds was investigated against five human cancer cell lines, confirming that 1 inhibits cell proliferation in A2058 human metastatic melanoma cells.

Mercury in a birch forest in SW Europe: Deposition flux by litterfall and pools in aboveground tree biomass and soils.

Atmospheric mercury (Hg) is largely assimilated by vegetation and subsequently transferred to the soil by litterfall, which highlights the role of forests as one of the largest global Hg sinks within terrestrial ecosystems. We assessed the pool of Hg in the aboveground biomass (leaves, wood, bark, branches and twigs), the Hg deposition flux through litterfall over two years (by sorting fallen biomass in leaves, twigs, reproductive structures and miscellaneous) and its accumulation in the soil profile in a deciduous forest dominated by Betula alba from SW Europe. The total Hg pool in the aboveground birch biomass was in the range 532-683 mg ha-1, showing the following distribution by plant tissues: well-developed leaves (171 mg ha-1) > twigs (160 mg ha-1) > bark (159 mg ha-1) > bole wood (145 mg ha-1) > fine branches (25 mg ha-1) > thick branches (24 mg ha-1) > newly sprouted leaves (20 mg ha-1). The total Hg deposition fluxes through litterfall were 15.4 and 11.7 µg m-2 yr-1 for the two years studied, with the greatest contribution coming from birch leaves (73 %). In the soil profile, the pool of Hg in the mineral soil (37.0 mg m-2) was an order of magnitude higher than in the organic horizons (1.0 mg m-2), mostly conditioned by parameters such as soil bulk density and thickness, total C and N contents and the presence of certain Al compounds.

Climate change related phenological decoupling in species belonging to the Betulaceae family.

Betulaceae species are anemophilous, and allergens from their pollen are a major cause of respiratory allergies in temperate areas where they are widely distributed. It is expected that, due to the strong influence of temperature on Betulaceae phenology, global warming will impact both the distribution and phenology of these species during the coming decades. This study examines potential decoupling of flowering and leafing phenophases in Betulaceae species (i.e. Alnus glutinosa, Betula pendula and Corylus avellana) over long-term (1951-2015) and as shorter (15-year) periods. Phenological phases for flowering and leaf unfolding of Betulaceae species from the Pan-European Phenology (PEP725) database were examined along with maximum and minimum daily temperature data for the periods September-October-November (SON), December-January-February (DJF) and March-April-May (MAM). Significant increases in temperature since 1951 have been recorded in the relevant chilling and forcing periods. Both flowering and leaf unfolding phenophases are advancing, but flowering is advancing faster than leaf unfolding. This is increasing the time between phenophases, although analysis of 15-year periods shows that the pattern of change was not constant. The results presented here represent the most comprehensive analysis of flowering and leaf unfolding phenophases of Betulaceae species using the PEP725 database to date. It is expected that these changes to Betulaceae phenology will continue and that global warming-related phenological decoupling will increase plant stress in Betulaceae populations in central Europe.

Plastome phylogenomics provide new perspective into the phylogeny and evolution of Betulaceae (Fagales).

BACKGROUND: Betulaceae is a relatively small but morphologically diverse family, with many species having important economic and ecological values. Although plastome structure of Betulaceae has been reported sporadically, a comprehensive exploration for plastome evolution is still lacking. Besides, previous phylogenies had been constructed based on limited gene fragments, generating unrobust phylogenetic framework and hindering further studies on divergence ages, biogeography and character evolution. Here, 109 plastomes (sixteen newly assembled and 93 previously published) were subject to comparative genomic and phylogenomic analyses to reconstruct a robust phylogeny and trace the diversification history of Betulaceae. RESULTS: All Betulaceae plastomes were highly conserved in genome size, gene order, and structure, although specific variations such as gene loss and IR boundary shifts were revealed. Ten divergent hotspots, including five coding regions (Pi > 0.02) and five noncoding regions (Pi > 0.035), were identified as candidate DNA barcodes for phylogenetic analysis and species delimitation. Phylogenomic analyses yielded high-resolution topology that supported reciprocal monophyly between Betula and Alnus within Betuloideae, and successive divergence of Corylus, Ostryopsis, and Carpinus-Ostrya within Coryloideae. Incomplete lineage sorting and hybridization may be responsible for the mutual paraphyly between Ostrya and Carpinus. Betulaceae ancestors originated from East Asia during the upper Cretaceous; dispersals and subsequent vicariance accompanied by historical environment changes contributed to its diversification and intercontinental disjunction. Ancestral state reconstruction indicated the acquisition of many taxonomic characters was actually the results of parallel or reversal evolution. CONCLUSIONS: Our research represents the most comprehensive taxon-sampled and plastome-level phylogenetic inference for Betulaceae to date. The results clearly document global patterns of plastome structural evolution, and established a well-supported phylogeny of Betulaceae. The robust phylogenetic framework not only provides new insights into the intergeneric relationships, but also contributes to a perspective on the diversification history and evolution of the family.

Fine-scale topographic influence on the spatial distribution of tree species diameter in old-growth beech (Fagus orientalis Lipsky.) forests, northern Iran.

The Hyrcanian forest in northern Iran is threatened by human use and encroachment and has suffered degradation in some areas. The forest has been declared a World Heritage Site and management in the region is shifting from timber production to conservation. There is considerable interest in developing a greater understanding of these diverse forest communities to inform forest management and multiple use plans to maintain the diversity and resilience of these forests. The Hyrcanian forest is characterized by a complex topography of catenas ranging up mountain slopes. Topographic gradients greatly influence microhabitat conditions which in turn impact tree distribution. To date there has been limited research on the impacts of this diverse topography on the spatial distribution of tree species and tree diameters in Hyrcanian forests. Such information is necessary to better understand the regional traits of tree diameters in these natural mixed temperate forests before forest management occurs. We examined the influence of the area's catena topography on the spatial pattern of tree species and on species stand structure in terms of tree diameter distribution. To quantify these dynamics, we conducted a complete enumeration inventory of all trees with dbh >12 cm within a 7.947 ha study area that included three C-shaped (concave) and three V-shaped (convex) catenas. Geostatistical variogram analysis and Clark and Evans aggregation index were utilized to study the spatial distribution of tree diameters. Beech, alder, hornbeam, linden and Persian maple exhibited clustered patterns, and sour cherry, ash, and oak exhibited random patterns. Geostatistical analysis clearly revealed the substantial influence of catena topography on the diameter distributions of alder and linden, more subtle influence on the diameter distributions of beech, and a possible influence on Persian maple, providing valuable insight into stand structure over neighborhood-based indices alone. Alder and linden both exhibited strong spatial structure in their diameter distributions (56% and 86%, respectively) where their diameter was strongly correlated with trees within 108 m and 83 m, respectively, sharing more similar diameters to each other than trees beyond that distance. Beech, maple, and hornbeam exhibited very weak if any spatial structure over short distances. These findings can be used to support the alignment of forest management practices in managed Hyrcanian forests with goals of protecting and maintaining biodiversity and sustainable forest ecosystems, and to inform geospatial modeling of species diameter distributions in areas where a complete stem-map is not feasible.

Genomic evidence for homoploid hybrid speciation between ancestors of two different genera.

Homoploid hybrid speciation (HHS) has been increasingly recognized as occurring widely during species diversification of both plants and animals. However, previous studies on HHS have mostly focused on closely-related species while it has been rarely reported or tested between ancestors of different genera. Here, we explore the likely HHS origin of Carpinus sect. Distegocarpus between sect. Carpinus and Ostrya in the family Betulaceae. We generate a chromosome-level reference genome for C. viminea of sect. Carpinus and re-sequence genomes of 44 individuals from the genera Carpinus and Ostrya. Our integrated analyses of all genomic data suggest that sect. Distegocarpus, which has three species, likely originates through HHS during the early divergence between Carpinus and Ostrya. Our study highlights the likelihood of an HHS event between ancestors of the extant genera during their initial divergences, which may have led to reticulate phylogenies at higher taxonomic levels.

Imaging Flow Cytometry as a Quick and Effective Identification Technique of Pollen Grains from Betulaceae, Oleaceae, Urticaceae and Asteraceae.

Despite the continuous and intensive development of laboratory techniques, a light microscope is still the most common tool used in pollen grains differentiation. However, microscopy is time-consuming and needs well-educated and experienced researchers. Other currently used techniques can be categorised as images and non-images analysis, but each has certain limitations. We propose a new approach to differentiate pollen grains using the Imaging Flow Cytometry (IFC) technique. It allows for high-throughput fluorescence data recording, which, in contrast to the standard FC, also enables real-time control of the results thanks to the possibility of digital image recording of cells flowing through the measuring capillary. The developed method allows us to determine the characteristics of the pollen grains population based on the obtained fluorescence data, using various combinations of parameters available in the IDEAS software, which can be analysed on different fluorescence channels. On this basis, we distinguished pollen grains both between and within different genera belonging to the Betulaceae, Oleaceae, Urticaceae and Asteraceae families. Thereby, we prove that the proposed methodology is sufficient for accurate, fast, and cost-effective identification and potentially can be used in the routine analysis of allergenic pollen grains.

Isolation and quantification of diarylheptanoids from European hornbeam (Carpinus betulus L.) and HPLC-ESI-MS/MS characterization of its antioxidative phenolics.

Detailed polyphenol profiling of European hornbeam (Carpinus betulus L.) bark, leaf, male and female catkin extracts was performed by high-performance liquid chromatography-diode array detection coupled to tandem mass spectrometry (HPLC-DAD-MS/MS). A total of 194 compounds were characterized and tentatively identified. Gallo- and ellagitannins dominated in the methanol extracts, while flavonol glycosides and methoxylated flavones prevailed in the ethyl acetate samples. In the quest for diarylheptanoids, twelve compounds were isolated by the combination of subsequent reversed-phase flash chromatographic and high-performance liquid chromatographic methods. The structural elucidation of the isolated components was performed by ultrahigh-performance liquid chromatography-Orbitrap mass spectrometry (UHPLC-Orbitrap-MS) as well as 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. Six known cyclic diarylheptanoids, together with a new compound were described in Carpinus betulus for the first time. The occurrence of a linear diarylheptanoid and a lignan has also been unprecedented in the genus Carpinus. Moreover, three known flavonol glycosides were isolated. Based on the identification of characteristic fragment ions, a new mass spectrometric fragmentation pathway for meta,meta-cyclophane-type diarylheptanoids was proposed. Quantities of the four major cyclic diarylheptanoids in European hornbeam were determined by a validated UHPLC-DAD method for the first time. The antioxidant properties of the extracts and the isolated compounds were assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Contribution of the individual constituents to the total radical scavenging activity of the samples was evaluated by an off-line DPPH-HPLC-DAD method. This allowed the identification of gallo- and ellagitannin derivatives as the constituents being primarily responsible for the antioxidant capacity of the extracts.

Molecular and morphological analyses clarify species delimitation in section Costatae and reveal Betula buggsii sp. nov. (sect. Costatae, Betulaceae) in China.

BACKGROUND AND AIMS: Delineating closely related and morphologically similar species is difficult. Here, we integrate morphology, genetics, ploidy and geography to resolve species and subspecies boundaries in four trees of section Costatae (genus Betula): Betula ashburneri, B. costata, B. ermanii and B. utilis, as well as multiple subspecies and polyploid races. METHODS: We genotyped 371 individuals (20-133 per species) from 51 populations at 15 microsatellite markers, as well as a subset of individuals, using restriction-site associated DNA sequencing and nuclear internal transcribed spacers. We determined the ploidy level of eight individuals using flow cytometry and characterized leaf variation for a subset of 109 individuals by morphometric analysis. KEY RESULTS: Integration of multiple lines of evidence suggested a series of revisions to the taxonomy of section Costatae. Betula costata and B. ermanii were found to be valid. Molecular and leaf morphology analyses revealed little differentiation between diploid B. albosinensis and some samples of B. utilis ssp. utilis. By contrast, other B. utilis ssp. utilis samples and ssp. albosinensis formed a morphological continuum but differed based on genetics. Specifically, B. utilis ssp. albosinensis was divided into two groups with group I genetically similar to B. utilis ssp. utilis and group II, a distinct cluster, proposed as the new diploid species Betula buggsii sp. nov. Phylogenomic analysis based on 2285 620 single nucleotide polymorphisms identified a well-supported monophyletic clade of B. buggsii. Morphologically, B. buggsii is characterized by elongated lenticels and a distinct pattern of bark peeling and may be geographically restricted to the Qinling-Daba Mountains. CONCLUSIONS: Our integrated approach identifies six taxa within section Costatae: B. ashburneri, B. buggsii, B. costata, B. utilis ssp. utilis, B. utilis ssp. albosinensis and B. ermanii. Our research demonstrates the value of an integrative approach using morphological, geographical, genetic and ploidy-level data for species delineation.

Potential distribution of the extremely endangered species Ostrya rehderiana (Betulaceae) in China under future climate change.

Global climate change is a major threat to biodiversity, which may increase the extinction risk of rare species, particularly those like Ostrya rehderiana Chun (Betulaceae) with very few remaining extant wild individuals. We aimed to estimate the potential distribution of O. rehderiana under climate change and to analyze possible relevant climatic factors. Maximum entropy (Maxent) was employed to model the potential distribution of O. rehderiana under present and future climate scenarios. Suitable habitat areas in different periods and the main contributing climate factors were identified using species distribution models. The minimum temperature in winter and precipitation seasonality were the principal climatic factors influencing the establishment of O. rehderiana. The proportion of high potential distribution area in China was 3.91% and would further shrink significantly under changing climate, especially reduce by 97% under high radiative forcing. The extinction risk of O. rehderiana would still be extraordinarily high under future climate scenarios. The Tianmu and Luoxiao Mountains would be the only potential refugia for O. rehderiana in the future. Special conservation efforts are urgently required to rescue extremely endangered species as O. rehderiana. We propose priorities for the conservation region and suggestions for conservation management strategies.

Comprehensive Transcriptome Analysis of Rare Carpinus putoensis Plants under NO2 stress.

We evaluated a transcriptome using high-throughput Illumina HiSeq sequencing and related it to the morphology, leaf anatomy, and physiological parameters of Carpinus putoensis putoensis under NO2 stress. The molecular mechanism of the C. putoensis NO2 stress response was evaluated using sequencing data. NO2 stress adversely affected the morphology, leaf anatomy, and total peroxidase (POD) activity. Through RNA-seq analysis, we used NCBI to compare the transcripts with nine databases and obtained their functional annotations. We annotated up to 2255 million clean Illumina paired-end RNA-seq reads, and 250,200 unigene sequences were assembled based on the resulting transcriptome data. More than 89% of the C. putoensis transcripts were functionally annotated. Under NO2 stress, 1119 genes were upregulated and 1240 were downregulated. According to the KEGG pathway and GO analyses, photosynthesis, chloroplasts, plastids, and the stimulus response are related to NO2 stress. Additionally, NO2 stress changed the expression of POD families, and the HPL2, HPL1, and POD genes exhibited high expression. The transcriptome analysis of C. putoensis leaves under NO2 stress supplies a reference for studying the molecular mechanism of C. putoensis resistance to NO2 stress. The given transcriptome data represent a valuable resource for studies on plant genes, which will contribute towards genome annotations during future genome projects.

Physiological and biochemical responses of two precious Carpinus species to high-concentration NO2 stress and their natural recovery.

Carpinus betulus and Carpinus putoensis are precious species in the world. Studies on the ecosystem function of the two species are rare. This study investigated the physiological and biochemical responses of C. betulus and C. putoensis to NO2 stress and their natural recovery. C. betulus and C. putoensis seedlings underwent fumigation with 12.0 mg/m3 NO2 for 0, 1, 6, 12, 24, 48, and 72 h, respectively. Then, the plants were allowed to recover at room temperature for 30 d. Physiological and biochemical changes in the leaves were compared between the two species. In terms of peroxidase (POD) activity, the damage response of C. betulus under NO2 stress appeared later than that of C. putoensis. The soluble protein content of C. betulus was noticeably higher than that of C. putoensis, and C. betulus exhibited more stable membrane lipoperoxidation. The tendency of the changes in nitrate reductase of C. betulus was less noticeable than that of C. putoensis. The variation amplitudes of N, K, Mg, Zn and Mn in the leaves of C. putoensis were greater than those of C. betulus. C. putoensis showed more sensitive metabolisms in response to NO2 stress compared with C. betulus. High-concentration NO2 caused damage to C. betulus and C. putoensis was reversible, and both species returned to normal growth via their own metabolism after 30-d recovery. The results of this study may provide useful reference data for quantitative assessment of the ecosystem function of C. betulus and C. putoensis and for their scientific application in urban greening.

Differences in survival and phenotypic traits of curly birch preserved by heterovegetative propagation: a case study from Central-East Europe.

Curly birch (Betula pendula Roth. var. carelica [Merklin] Hämet-Ahti) is a disappearing representative of the Betula genus facing a regeneration failure in a large part of its natural distribution in Europe. The unique long-term study of clonal replications originating in heterogeneous environments enabled the evaluation of long-term survival and phenotypic stability of progenies in seed orchard to assess the conservation and commercial potential of heterovegetative propagation. Seventy-eight geographic sources (95 clone origins) representing the south distribution edge in East-Central Europe were analysed for species variation in survival, growth form, bark colour, and stem quality of parent trees and their vegetative progeny, and the effects of four parental site origin characteristics. The survival rate was 73% after 28-33 years of growth. Retention of curly-grained wood was high, the curly-grained wood structure is heritable and thus clonally efficiently achievable (only 3.5% of grafted individuals showed no occurrence of figured wood structure). The phenotypic expression of curliness manifested on the trunks as bulges, stem growth forms (tree/shrub) and stem technical quality showed a lower degree of stability (coincidence) between the parent trees and heterovegatively propagated progenies. Despite this, the conservation potential of seed orchard is very high, especially when stabilization of the stem growth forms affecting the survival and commercial value of progenies can be probably achieved by a more careful selection of scions. Overall, heterovegetative orchards seem to be a very promising method for the long-term conservation of curly birch populations, which, in addition to their great biological and ecological value, have considerable commercial potential.

Cryphonectria carpinicola sp. nov. Associated with hornbeam decline in Europe.

Since the early 2000s, reports on declining hornbeam trees (Carpinus betulus) are spreading in Europe. Two fungi are involved in the decline phenomenon: One is Anthostoma decipiens, but the other etiological agent has not been identified yet. We examined the morphology, phylogenetic position, and pathogenicity of yellow fungal isolates obtained from hornbeam trees from Austria, Georgia and Switzerland, and compared data with disease reports from northern Italy documented since the early 2000s. Results demonstrate distinctive morphology and monophyletic status of Cryphonectria carpinicola sp. nov. as etiological agent of the European hornbeam decline. Interestingly, the genus Cryphonectria splits into two major clades. One includes Cry. carpinicola together with Cry. radicalis, Cry. decipiens and Cry. naterciae from Europe, while the other comprises species known from Asia-suggesting that the genus Cryphonectria has developed at two evolutionary centres, one in Europe and Asia Minor, the other in East Asia. Pathogenicity studies confirm that Car. betulus is a major host species of Cry. carpinicola. This clearly distinguished Cry. carpinicola from other Cryphonectria species, which mainly occur on Castanea and Quercus.