Genomic dissection of additive and non-additive genetic effects and genomic prediction in an open-pollinated family test of Japanese larch.

Genomic dissection of genetic effects on desirable traits and the subsequent use of genomic selection hold great promise for accelerating the rate of genetic improvement of forest tree species. In this study, a total of 661 offspring trees from 66 open-pollinated families of Japanese larch (Larix kaempferi (Lam.) Carrière) were sampled at a test site. The contributions of additive and non-additive effects (dominance, imprinting and epistasis) were evaluated for nine valuable traits related to growth, wood physical and chemical properties, and competitive ability using three pedigree-based and four Genomics-based Best Linear Unbiased Predictions (GBLUP) models and used to determine the genetic model. The predictive ability (PA) of two genomic prediction methods, GBLUP and Reproducing Kernel Hilbert Spaces (RKHS), was compared. The traits could be classified into two types based on different quantitative genetic architectures: for type I, including wood chemical properties and Pilodyn penetration, additive effect is the main source of variation (38.20-67.46%); for type II, including growth, competitive ability and acoustic velocity, epistasis plays a significant role (50.76-91.26%). Dominance and imprinting showed low to moderate contributions (< 36.26%). GBLUP was more suitable for traits of type I (PAs = 0.37-0.39 vs. 0.14-0.25), and RKHS was more suitable for traits of type II (PAs = 0.23-0.37 vs. 0.07-0.23). Non-additive effects make no meaningful contribution to the enhancement of PA of GBLUP method for all traits. These findings enhance our current understanding of the architecture of quantitative traits and lay the foundation for the development of genomic selection strategies in Japanese larch.

Phylogeny and expression patterns of ERF genes that are potential reproductive inducers in hybrid larch.

BACKGROUND: Larch is an important component of northern forests and a major cultivated tree species in restoration of forest cover using improved seed material. In recent years, the continuous low seed production has severely affected the production of improved variety seedlings and natural regeneration. However, research on the reproductive growth of gymnosperms is extremely scarce. RESULTS: In this study, based on differential transcriptome analysis of two asexual reproductive phases, namely high-yield and low-yield, we further screened 5 ERF family genes that may affect the reproductive development of larch. We analyzed their genetic relationships and predicted their physicochemical properties. The expression patterns of these genes were analyzed in different tissues, developmental stages, hormone treatments, and environmental conditions in hybrid larch. CONCLUSION: The results showed that all 5 genes were induced by low temperature and ABA, and their expression patterns in different tissues suggested a suppressive role in the development of female cones in larch. Among them, LkoERF3-like1 and LkoERF071 may be involved in the flowering age pathway. This study enriches the scarce research on reproductive development in gymnosperms and provides a theoretical basis and research direction for regulating the reproductive development of larch in seed orchards.

H2O2 Significantly Affects Larix kaempferi × Larix olgensis Somatic Embryogenesis.

Larch is widely distributed throughout the world and is an important species for timber supply and the extraction of industrial raw materials. In recent years, the hybrid breeding of Larix kaempferi and Larix olgensis has shown obvious heterosis in quick-growth, stress resistance and wood properties. However, its growth and development cycle is too long to meet general production needs. In order to shorten the breeding cycle, we have for the first time successfully established and optimized a somatic embryogenesis system for Larix kaempferi × Larix olgensis. We found that the highest rate of embryonal-suspensor mass (ESM) induction was observed when late cotyledonary embryos were used as explants. The induced ESMs were subjected to stable proliferation, after which abscisic acid (ABA) and polyethylene glycol (PEG) were added to successfully induce somatic embryos. Treatment with PEG and ABA was of great importance to somatic embryo formation and complemented each other's effect. ABA assisted embryo growth, whereas PEG facilitated the formation of proembryo-like structures. On top of this, we studied in more detail the relationship between redox homeostasis and the efficiency of somatic embryogenesis (frequency of ESM induction). During subculture, we observed the gradual formation of three distinct types of ESM. The Type I ESM is readily able to form somatic embryos. In contrast to type I, the type III ESM suffers from severe browning, contains a higher level of hydrogen peroxide (H2O2) and demonstrates a decreased ability to form somatic embryos. External treatment with H2O2 decreased the somatic embryogenesis efficiency of Type I and type III ESMs, or the higher the exogenous H2O2 content, the lower the resulting somatic embryogenesis efficiency. We found that treatment with the H2O2 scavenger DMTU (dimethylthiourea) could significantly increase the somatic embryogenesis efficiency of the type III ESM, as a result of a decline in endogenous H2O2 content. Overall, these findings have contributed to setting up a successful somatic embryogenesis system for larch production.

Cambium Reactivation Is Closely Related to the Cell-Cycle Gene Configuration in Larix kaempferi.

Dormancy release and reactivation in temperate trees are mainly controlled by temperature and are affected by age, but the underlying molecular mechanisms are still unclear. In this study, we explored the effects of low temperatures in winter and warm temperatures in spring on dormancy release and reactivation in Larix kaempferi. Further, we established the relationships between cell-cycle genes and cambium cell division. The results showed that chilling accelerated L. kaempferi bud break overall, and the longer the duration of chilling is, the shorter the bud break time is. After dormancy release, warm temperatures induced cell-cycle gene expression; when the configuration value of the cell-cycle genes reached 4.97, the cambium cells divided and L. kaempferi reactivated. This study helps to predict the impact of climate change on wood production and provides technical support for seedling cultivation in greenhouses.

UAV Photogrammetry for Estimating Stand Parameters of an Old Japanese Larch Plantation Using Different Filtering Methods at Two Flight Altitudes.

Old plantations are iconic sites, and estimating stand parameters is crucial for valuation and management. This study aimed to estimate stand parameters of a 115-year-old Japanese larch (Larix kaempferi (Lamb.) Carrière) plantation at the University of Tokyo Hokkaido Forest (UTHF) in central Hokkaido, northern Japan, using unmanned aerial vehicle (UAV) photogrammetry. High-resolution RGB imagery was collected using a DJI Matrice 300 real-time kinematic (RTK) at altitudes of 80 and 120 m. Structure from motion (SfM) technology was applied to generate 3D point clouds and orthomosaics. We used different filtering methods, search radii, and window sizes for individual tree detection (ITD), and tree height (TH) and crown area (CA) were estimated from a canopy height model (CHM). Additionally, a freely available shiny R package (SRP) and manually digitalized CA were used. A multiple linear regression (MLR) model was used to estimate the diameter at breast height (DBH), stem volume (V), and carbon stock (CST). Higher accuracy was obtained for ITD (F-score: 0.8-0.87) and TH (R2: 0.76-0.77; RMSE: 1.45-1.55 m) than for other stand parameters. Overall, the flying altitude of the UAV and selected filtering methods influenced the success of stand parameter estimation in old-aged plantations, with the UAV at 80 m generating more accurate results for ITD, CA, and DBH, while the UAV at 120 m produced higher accuracy for TH, V, and CST with Gaussian and mean filtering.

Genetic Adaptation of Siberian Larch (Larix sibirica Ledeb.) to High Altitudes.

Forest trees growing in high altitude conditions offer a convenient model for studying adaptation processes. They are subject to a whole range of adverse factors that are likely to cause local adaptation and related genetic changes. Siberian larch (Larix sibirica Ledeb.), whose distribution covers different altitudes, makes it possible to directly compare lowland with highland populations. This paper presents for the first time the results of studying the genetic differentiation of Siberian larch populations, presumably associated with adaptation to the altitudinal gradient of climatic conditions, based on a joint analysis of altitude and six other bioclimatic variables, together with a large number of genetic markers, single nucleotide polymorphisms (SNPs), obtained from double digest restriction-site-associated DNA sequencing (ddRADseq). In total, 25,143 SNPs were genotyped in 231 trees. In addition, a dataset of 761 supposedly selectively neutral SNPs was assembled by selecting SNPs located outside coding regions in the Siberian larch genome and mapped to different contigs. The analysis using four different methods (PCAdapt, LFMM, BayeScEnv and RDA) revealed 550 outlier SNPs, including 207 SNPs whose variation was significantly correlated with the variation of some of environmental factors and presumably associated with local adaptation, including 67 SNPs that correlated with altitude based on either LFMM or BayeScEnv and 23 SNPs based on both of them. Twenty SNPs were found in the coding regions of genes, and 16 of them represented non-synonymous nucleotide substitutions. They are located in genes involved in the processes of macromolecular cell metabolism and organic biosynthesis associated with reproduction and development, as well as organismal response to stress. Among these 20 SNPs, nine were possibly associated with altitude, but only one of them was identified as associated with altitude by all four methods used in the study, a nonsynonymous SNP in scaffold_31130 in position 28092, a gene encoding a cell membrane protein with uncertain function. Among the studied populations, at least two main groups (clusters), the Altai populations and all others, were significantly genetically different according to the admixture analysis based on any of the three SNP datasets as follows: 761 supposedly selectively neutral SNPs, all 25,143 SNPs and 550 adaptive SNPs. In general, according to the AMOVA results, genetic differentiation between transects or regions or between population samples was relatively low, although statistically significant, based on 761 neutral SNPs (FST = 0.036) and all 25,143 SNPs (FST = 0.017). Meanwhile, the differentiation based on 550 adaptive SNPs was much higher (FST = 0.218). The data showed a relatively weak but highly significant linear correlation between genetic and geographic distances (r = 0.206, p = 0.001).

Adaptation of Betula pendula Roth., Pinus sylvestris L., and Larix decidua Mill. to environmental stress caused by tailings waste highly contaminated by trace elements.

The seedlings of some tree species can successfully develop in areas polluted by heavy metals. Research on such species is important in order to explore the possibility of introducing tree species for the permanent biological stabilization and reclamation of post-flotation tailings, especially after the final recycling of trace metals, but where concentrations remain much higher than in natural soils. To better understand the adaptation and reaction of Betula pendula Roth., Pinus sylvestris L., and Larix decidua Mill. seedlings to heavy metals pollution caused by tailings waste highly contaminated by trace elements: 1) the relationships between the concentration of heavy metals in the soil substrate, the efficiency of heavy metal ions accumulation in plant organs, and the biometric parameters of the seedlings; and 2) the threshold content of heavy metals in the roots above which the plant physiological response is triggered was determined. We assume that there are certain limit concentrations of heavy metals in the soil and fine roots, which depend on the tree species and beyond which the plant responds strongly to stressThe obtained results showed that Betula is a suitable species for the phytostabilization of post-flotation tailings due to its rapid growth rate and production of root biomass. The accumulation of metals in Betula roots was found to be much greater than in Pinus and Larix. Despite the high concentrations of heavy metals in the prepared substrates, there was only a slight transfer of these elements to the aboveground parts of the plant. At high soil concentrations, the heavy metals adversely affected the cellular and physiological processes of plants. In plants growing in such conditions, the activity of the antioxidant system depended both on the species and organ of the plant, as well as on the type and metal concentration.

Tree growth is connected with distribution and warming-induced degradation of permafrost in southern Siberia.

Hemiboreal and boreal forests growing at the southern margin of the permafrost distribution are vulnerable to climate warming. However, how climate warming threatens the growth of dominant tree species that are distributed on permafrost remains to be determined, particularly in synchrony with warming-induced permafrost degradation. Tree growth in the permafrost region of southern Siberia was hypothesized to be highly sensitive to temperature increasing and warming-induced permafrost degradation. To test this hypothesis, we sampled the tree ring width of 535 trees of dominant species, larch (including Larix gmelinii and L. sibirica) and white birch (Betula platyphylla), in ten hemiboreal to boreal forest plots within different permafrost zones. The relationships between the tree ring basal area index (BAI) and temperature, precipitation, and the Palmer drought severity index (PDSI) were compared among plots located in two permafrost zones. In the isolated permafrost zone, white birch grows better than larch and is not drought-stressed (p < .05). We suggest that the deep-rooted white birch benefits from the water from thawing permafrost, while the growth of the shallow-rooted larch is stressed by drought. In the sporadic discontinuous permafrost zone, both white birch and larch benefited from permafrost melting, but the sensitivity of larch growth to PDSI is still significant (p < .05), indicating drought is still an important climatic factor limiting the growth of larch. Our results imply that the permafrost degradation caused by climate warming affects tree growth by creating the root layer additional water source. In the future, it is necessary to focus on monitoring permafrost degradation to better predict forest dynamics at the southern margin of the permafrost distribution.

LaDAL1 Coordinates Age and Environmental Signals in the Life Cycle of Larix kaempferi.

Perennial woody plants are long-lived, and their life-cycle events occur in order in each generation, but what drives the occurrence and restart of these events in their offspring is unknown. Based on its age-dependent expression pattern and function, Larix kaempferi DEFICIENS-AGAMOUS-LIKE 1 (LaDAL1), a MADS transcription factor has been suggested to be a time recorder and life-cycle event coordinator. Here, we studied the dynamic spatiotemporal expression pattern of LaDAL1 in the life cycle of L. kaempferi to analyze the molecular mechanism of life-cycle progression. In full view of the life cycle, LaDAL1 transcription was related with life-cycle progression, and its transcript level increased sharply from age 3 to 5 years, which might be the molecular characteristic of the vegetative phase change, and then stayed at a high level. During sexual reproduction, LaDAL1 transcript level decreased sequentially during meiosis and embryogenesis, suggesting that meiosis rapidly lowers the age signal, and after fertilization, the age signal was reset to "0" with the embryogenesis. When a seed germinates, the next generation restarts, and the age is re-counted. Altogether, these results not only provide important and novel insights into the life-cycle progression and transgeneration in perennial woody plants, but also advance our understanding of age recording.

Antimicrobial Activity and Wound-Healing Capacity of Birch, Beech and Larch Bark Extracts.

Bark is a major by-product of woodworking industries. The contents of several wood species are known to harbor antimicrobial, antiviral, anti-inflammatory and wound-healing capacities. The aim of this work was to identify beneficial properties of Austrian larch, birch and beech bark extracts for their potential usage as additives or active ingredients in dermatological applications. Bacterial agar diffusion assay and resazurin-based broth microdilution assay were used to evaluate anti-bacterial activity. To gain more insight into the cellular response to bark extracts, viability-, scratch-assays and ELISAs were performed. Birch and beech extracts showed strong antimicrobial activities against Gram-positive bacteria, including Cutibacterium acnes, Staphylococcus epidermidis and MRSA. Wound closure was enhanced with birch and beech extracts as compared to controls in the scratch-assays. Whereas beneficial properties of birch bark components have previously been described, the similar effects of beech extracts are novel. The combined positive effect on wound-healing and antimicrobial activity has great potential for the treatment of various skin diseases, including acne in future dermal applications.

A 168-year temperature recording based on tree rings and latitude differences in temperature changes in northeast China.

A ring-width series was developed from Dahurian larch (Larix gmelinii) in the northeastern forest area of Inner Mongolia, China. By analyzing the relationships between tree-ring data and climate records, an August-September mean maximum temperature (T89) series during 1845 and 2012 was reconstructed based on a simple linear regression equation. This reconstructed series explained 40.9% variance of the observed temperature from 1959 to 2012. The reconstructed T89 series was consistent with the historical disaster events caused by extreme climate (e.g., flood, frost disaster, and cold damage). Besides, the temperature comparisons showed that the year in which the warm months (April-September) in northeast China began to warm up has latitude differences. It started with a gradual delay from north to south, starting 1980 in the south region, after 1950 AD in the central region and after 1940 in the north region. Our study can enrich high-resolution temperature series in Northeast China and help clarify the characteristic of recent warming in northeast China.

Physiological and anatomical responses to drought stress differ between two larch species and their hybrid.

KEY MESSAGE: Hybrid saplings were more reactive to soil water deficit than Japanese and European larch. European larch had hydraulically safer wood and anisohydric behavior, Japanese and hybrid larch showed isohydric strategy. ABSTRACT: Deciduous larch species could be an alternative to evergreen conifers in reforestation, but little is known about drought sensitivity of their saplings. The effect of an experimental drought on hydraulics and quantitative wood anatomy was tested on saplings of European larch (EL, Larix decidua), Japanese larch (JL, Larix kaempferi) and their hybrid (HL). Across species, biomass, transpiration rate and relative water content were higher in controls than in drought stressed trees, but transpiration efficiency was lower. JL had the highest transpiration efficiency under drought, and EL the lowest, coinciding with slower growth of EL. Wood of EL formed before drought was hydraulically safer as shown by higher wall/lumen ratio and lower pit cavity area. EL neither had a significant increase in transpiration efficiency nor a reduction in transpiration rate under drought, suggesting that the stomata remained open under soil water deficit. HL saplings were the most reactive to water shortage, indicated by intra-annual density fluctuations and a decrease in relative water content of the sapwood. Significant reduction in transpiration by HL suggested a higher stomatal sensitivity, while the same leaf surface area was maintained and radial growth was still similar to its best parent, the JL. The latter showed a significantly lower leaf surface area under drought than controls. EL, with its hydraulically safer wood, followed an anisohydric behavior, while JL and HL revealed an isohydric strategy. Altogether, our results suggest species dependent acclimations to drought stress, whereby HL followed the strategy of JL rather than that of EL. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00468-021-02129-4.

Stepwise large genome assembly approach: a case of Siberian larch (Larix sibirica Ledeb).

BACKGROUND: De novo assembling of large genomes, such as in conifers (~ 12-30 Gbp), which also consist of ~ 80% of repetitive DNA, is a very complex and computationally intense endeavor. One of the main problems in assembling such genomes lays in computing limitations of nucleotide sequence assembly programs (DNA assemblers). As a rule, modern assemblers are usually designed to assemble genomes with a length not exceeding the length of the human genome (3.24 Gbp). Most assemblers cannot handle the amount of input sequence data required to provide sufficient coverage needed for a high-quality assembly. RESULTS: An original stepwise method of de novo assembly by parts (sets), which allows to bypass the limitations of modern assemblers associated with a huge amount of data being processed, is presented in this paper. The results of numerical assembling experiments conducted using the model plant Arabidopsis thaliana, Prunus persica (peach) and four most popular assemblers, ABySS, SOAPdenovo, SPAdes, and CLC Assembly Cell, showed the validity and effectiveness of the proposed stepwise assembling method. CONCLUSION: Using the new stepwise de novo assembling method presented in the paper, the genome of Siberian larch, Larix sibirica Ledeb. (12.34 Gbp) was completely assembled de novo by the CLC Assembly Cell assembler. It is the first genome assembly for larch species in addition to only five other conifer genomes sequenced and assembled for Picea abies, Picea glauca, Pinus taeda, Pinus lambertiana, and Pseudotsuga menziesii var. menziesii.

Siberian larch (Larix sibirica Ledeb.) chloroplast genome and development of polymorphic chloroplast markers.

BACKGROUND: The main objectives of this study were sequencing, assembling, and annotation of chloroplast genome of one of the main Siberian boreal forest tree conifer species Siberian larch (Larix sibirica Ledeb.) and detection of polymorphic genetic markers - microsatellite loci or simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). RESULTS: We used the data of the whole genome sequencing of three Siberian larch trees from different regions - the Urals, Krasnoyarsk, and Khakassia, respectively. Sequence reads were obtained using the Illumina HiSeq2000 in the Laboratory of Forest Genomics at the Genome Research and Education Center of the Siberian Federal University. The assembling was done using the Bowtie2 mapping program and the SPAdes genomic assembler. The genome annotation was performed using the RAST service. We used the GMATo program for the SSRs search, and the Bowtie2 and UGENE programs for the SNPs detection. Length of the assembled chloroplast genome was 122,561 bp, which is similar to 122,474 bp in the closely related European larch (Larix decidua Mill.). As a result of annotation and comparison of the data with the existing data available only for three larch species - L. decidua, L. potaninii var. chinensis (complete genome 122,492 bp), and L. occidentalis (partial genome of 119,680 bp), we identified 110 genes, 34 of which represented tRNA, 4 rRNA, and 72 protein-coding genes. In total, 13 SNPs were detected; two of them were in the tRNA-Arg and Cell division protein FtsH genes, respectively. In addition, 23 SSR loci were identified. CONCLUSIONS: The complete chloroplast genome sequence was obtained for Siberian larch for the first time. The reference complete chloroplast genomes, such as one described here, would greatly help in the chloroplast resequencing and search for additional genetic markers using population samples. The results of this research will be useful for further phylogenetic and gene flow studies in conifers.

Accumulation of heavy metals and biochemical responses in Siberian larch needles in urban area.

Active urbanization processes exacerbate environmental problems associated with industrial pollution in cities. Urban greening helps reduce level of air pollution and improve microclimate. Selection sensitive plant species (indicators of pollution), and the resistant species (decrease the level pollution) is acute in many countries. The aim of the present work was to establish concentrations of heavy metals (Fe, Mn, Zn, Cu, Ni, Cr, Pb, Co and Cd) in the Siberian larch needles grown in various urban land-use (functional) zones of Ulan-Ude (Russia), as well as to determinate and compare the levels of some biochemical compounds. Based on index of soil contamination, the highest heavy metal pollution was found in the highway and industrial zones. The index of biogeochemical transformation of the needle elements composition ranged from 5.1 (minimal level) to 32.2 (strong level). The most polluted sites were along highways, where Fe, Zn, Cu, Cr, Ni, Pb, and Cd concentration in the needles were up to 2.5-7.7 times than background values. An important role in the protective system of larch is played by pigments, especially Chl b and carotenoids. Their content in the needles is increased by 1.3-2.2 times. Ratio Chl a/b and ∑Chl/carotenoids decrease as compared to background level; in the first case-due to increase of Chl b content, in the second case-increase of carotenoids level. Highest concentrations of proline, condensed tannins and peroxidase activity were found in needles from urban zones connected with high traffic and industrial emission. Based on the Air Pollution Tolerance Index Siberian larch should be considered sensitive species to air pollution and can be recommended as bioindicator.

A 400-year reconstruction of wintertime Arctic sea-ice extent using a high-elevation, mid-latitude tree-ring record.

We examined relationships between wintertime Arctic sea-ice extent (ASIE) and radial growth rates of alpine larch trees (LALY) in the northern Rockies, USA, during 1979-2015 (r = - 0.71, p < 0.01) and reconstructed ASIE fluctuations from 1613 to 2015. Wintertime ASIE corresponds with summertime mid-latitude upper-level atmospheric flow patterns with ridging (troughing) and warmer (cooler) conditions prevailing in the Rockies when ASIE are below (above) average. In turn, warmer (cooler) summertime conditions favor (disfavor) growth of LALY, thus "recording" interannual variations in ASIE. Both 1000 hPa temperatures and 500 hPa geopotential heights during years of anomalously high or low radial growth negatively correspond with ASIE, suggesting that a disjunct spatial influence on atmospheric conditions may be associated with interannual variability of ASIE. Reconstructed ASIE values show that 1996-2015 was the lowest 20-year period on record, but the mean value is not significantly different from six other distinct periods of below-average ASIE.

Effects of exogenous organic acids on the resistance of Changbai larch (Larix olgensis) seedlings to mixed Pb and Cd stress.

Mixed Pb and Cd soil contamination is an issue in Northeast China. We examined the effects of exogenous organic acids on the resilience of Changbai larch (Larix olgensis) seedlings, a pioneering forestry species in afforestation and vegetation restoration in Northeast China, under such stress. Mixed Pb and Cd stress led to significantly higher Pb and Cd content in the leaves and fine roots, malondialdehyde content in the leaves, superoxide dismutase activity, and soluble protein content in the leaves. Lower biomass of the roots, stems, and leaves was observed, with the roots showing the sharpest reduction in biomass. However, the application of organic acids mitigated or reversed these effects. This was most pronounced following treatment with 0.2 mmol·L-1 or 1.0 mmol·L-1 organic acids for 20 days. Citric acid had the greatest positive effect compared with succinic acid and oxalic acid. We suggest that exogenous organic acids have the potential to alleviate Pb and Cd-induced oxidation injury symptoms in Changbai larch seedlings, and may enhance resilience to mixed Pb and Cd stress.

Lifting Wavelet Transform De-noising for Model Optimization of Vis-NIR Spectroscopy to Predict Wood Tracheid Length in Trees.

The data analysis of visible-near infrared (Vis-NIR) spectroscopy is critical for precise information extraction and prediction of fiber morphology. The objectives of this study were to discuss the de-noising of Vis-NIR spectra, taken from wood, to improve the prediction accuracy of tracheid length in Dahurian larch wood. Methods based on lifting wavelet transform (LWT) and local correlation maximization (LCM) algorithms were developed for optimal de-noising parameters and partial least squares (PLS) was employed as the prediction method. The results showed that: (1) The values of tracheid length in the study were generally high and had a great positive linear correlation with annual rings (R = 0.881), (2) the optimal de-noising parameters for larch wood based Vis-NIR spectra were Daubechies-2 (db2) mother wavelet with 4 decomposition levels while using a global fixed hard threshold based on LWT, and (3) the Vis-NIR model based on the optimal LWT de-noising parameters ( R c 2 = 0.834, RMSEC = 0.262, RPD c = 2.454) outperformed those based on the LWT coupled with LCM algorithm (LWT-LCM) ( R c 2 = 0.816, RMSEC = 0.276, RPD c = 2.331) and raw spectra ( R c 2 = 0.822, RMSEC = 0.271, RPD c = 2.370). Thus, the selection of appropriate LWT de-noising parameters could aid in extracting a useful signal for better prediction accuracy of tracheid length.

Preparation and Characterization of Antioxidative and UV-Protective Larch Bark Tannin/PVA Composite Membranes.

In order to prepare functional materials for antioxidant and ultraviolet (UV)-protective green food packaging, condensed tannin, previously extracted from larch bark, was mixed with polyvinyl alcohol (PVA), and then the mixture was used to cast composite membranes. An antioxidative assay using 1,1-diphenyl-2-picrylhydrazyl (DPPH)-a free radical scavenger-and starch⁻potassium iodide oxidation⁻discoloration analyses showed that the composite membranes have good antioxidative activities. The low UV transmission and protective effect of the composite films on vitamin E indicated the UV protection ability of the composite membranes. Both larch bark tannin and PVA are rich in hydroxyl groups; scanning electron microscopy analysis demonstrated their compatibility. Also, the mechanical and crystallization properties of the composite membranes did not significantly decrease with the addition of larch bark tannin.

Soil warming opens the nitrogen cycle at the alpine treeline.

Climate warming may alter ecosystem nitrogen (N) cycling by accelerating N transformations in the soil, and changes may be especially pronounced in cold regions characterized by N-poor ecosystems. We investigated N dynamics across the plant-soil continuum during 6 years of experimental soil warming (2007-2012; +4 °C) at a Swiss high-elevation treeline site (Stillberg, Davos; 2180 m a.s.l.) featuring Larix decidua and Pinus uncinata. In the soil, we observed considerable increases in the NH4+ pool size in the first years of warming (by >50%), but this effect declined over time. In contrast, dissolved organic nitrogen (DON) concentrations in soil solutions from the organic layer increased under warming, especially in later years (maximum of +45% in 2012), suggesting enhanced DON leaching from the main rooting zone. Throughout the experimental period, foliar N concentrations showed species-specific but small warming effects, whereas δ15 N values showed a sustained increase in warmed plots that was consistent for all species analysed. The estimated total plant N pool size at the end of the study was greater (+17%) in warmed plots with Pinus but not in those containing Larix, with responses driven by trees. Irrespective of plot tree species identity, warming led to an enhanced N pool size of Vaccinium dwarf shrubs, no change in that of Empetrum hermaphroditum (dwarf shrub) and forbs, and a reduction in that of grasses, nonvascular plants, and fine roots. In combination, higher foliar δ15 N values and the transient response in soil inorganic N indicate a persistent increase in plant-available N and greater cumulative plant N uptake in warmer soils. Overall, greater N availability and increased DON concentrations suggest an opening of the N cycle with global warming, which might contribute to growth stimulation of some plant species while simultaneously leading to greater N losses from treeline ecosystems and possibly other cold biomes.