Phytochemical Characterization, Antioxidant, and Antimicrobial Activity of the Vegetative Buds from Romanian Spruce, Picea abies (L.) H. Karst.

This study aims to investigate the vegetative buds from Picea abies (spruce), naturally found in a central region of Romania, through a comprehensive analysis of the chemical composition to identify bioactive compounds responsible for pharmacological properties. Using HPLC/derivatization technique of GC-MS and quantitative spectrophotometric assays, the phenolic profile, and main components of an ethanolic extract from the buds were investigated. The essential oil was characterized by GC-MS. Moreover, the antioxidant activity with the DPPH method, and the antimicrobial activity were tested. Heavy metal detection was performed by graphite furnace atomic absorption spectrometry. The main components of the alcoholic extract were astragalin, quercetin, kaempferol, shikimic acid, and quinic acid. A total content of 25.32 ± 2.65 mg gallic acid equivalent per gram of dry plant (mg GAE/g DW) and of 10.54 ± 0.083 mg rutin equivalents/g of dry plant (mg RE/g DW) were found. The essential oil had D-limonene, α-cadinol, δ-cadinene, 13-epimanool, and δ-3-carene as predominant components. The spruce vegetative buds exhibited significant antioxidant activity (IC50 of 53 µg/mL) and antimicrobial effects against Staphylococcus aureus. Furthermore, concentrations of heavy metals Pb and Cd were below detection limits, suggesting that the material was free from potentially harmful contaminants. The results confirmed the potential of this indigenous species to be used as a source of compounds with pharmacological utilities.

Climate of origin shapes variations in wood anatomical properties of 17 Picea species.

BACKGROUND: Variations in hydraulic conductivity may arise from species-specific differences in the anatomical structure and function of the xylem, reflecting a spectrum of plant strategies along a slow-fast resource economy continuum. Spruce (Picea spp.), a widely distributed and highly adaptable tree species, is crucial in preventing soil erosion and enabling climate regulation. However, a comprehensive understanding of the variability in anatomical traits of stems and their underlying drivers in the Picea genus is currently lacking especially in a common garden. RESULTS: We assessed 19 stem economic properties and hydraulic characteristics of 17 Picea species grown in a common garden in Tianshui, Gansu Province, China. Significant interspecific differences in growth and anatomical characteristics were observed among the species. Specifically, xylem hydraulic conductivity (Ks) and hydraulic diameter exhibited a significant negative correlation with the thickness to span ratio (TSR), cell wall ratio, and tracheid density and a significant positive correlation with fiber length, and size of the radial tracheid. PCA revealed that the first two axes accounted for 64.40% of the variance, with PC1 reflecting the trade-off between hydraulic efficiency and mechanical support and PC2 representing the trade-off between high embolism resistance and strong pit flexibility. Regression analysis and structural equation modelling further confirmed that tracheid size positively influenced Ks, whereas the traits DWT, D_r, and TSR have influenced Ks indirectly. All traits failed to show significant phylogenetic associations. Pearson's correlation analysis demonstrated strong correlations between most traits and longitude, with the notable influence of the mean temperature during the driest quarter, annual precipitation, precipitation during the wettest quarter, and aridity index. CONCLUSIONS: Our results showed that xylem anatomical traits demonstrated considerable variability across phylogenies, consistent with the pattern of parallel sympatric radiation evolution and global diversity in spruce. By integrating the anatomical structure of the stem xylem as well as environmental factors of origin and evolutionary relationships, our findings provide novel insights into the ecological adaptations of the Picea genus.

Forest management positively reshapes the phyllosphere bacterial community and improves community stability.

Research has shown that forest management can improve the post-drought growth and resilience of Qinghai spruce in the eastern Qilian Mountains, located on the northeastern Tibetan Plateau. However, the impact of such management on the tree-associated phyllosphere microbiome is not yet fully understood. This study provides new evidence of positive forest management effects on the phyllosphere microbiome after extreme drought, from the perspectives of community diversity, structure, network inference, keystone species, and assembly processes. In managed Qinghai spruce forest, the α-diversity of the phyllosphere bacterial communities increased, whereas the ß-diversity decreased. In addition, the phyllosphere bacterial community became more stable and resistant, yet less complex, following forest management. Keystone species inferred from a bacterial network also changed under forest management. Furthermore, forest management mediated changes in community assembly processes, intensifying the influence of determinacy, while diminishing that of stochasticity. These findings support the hypothesis that management can re-assemble the phyllosphere bacterial community, enhance community stability, and ultimately improve tree growth. Overall, the study highlights the importance of forest management on the phyllosphere microbiome and furnishes new insights into forest conservation from the perspective of managing microbial processes and effects.

Probing the complexity of wood with computer vision: from pixels to properties.

We use data produced by industrial wood grading machines to train a machine learning model for predicting strength-related properties of wood lamellae from colour images of their surfaces. The focus was on samples of Norway spruce (Picea abies) wood, which display visible fibre pattern formations on their surfaces. We used a pre-trained machine learning model based on the residual network ResNet50 that we trained with over 15 000 high-definition images labelled with the indicating properties measured by the grading machine. With the help of augmentation techniques, we were able to achieve a coefficient of determination (R2) value of just over 0.9. Considering the ever-increasing demand for construction-grade wood, we argue that computer vision should be considered a viable option for the automatic sorting and grading of wood lamellae in the future.

Species-specific responses of C and N allocation to N addition: evidence from dual 13C and 15N labeling in three tree species.

Soil nitrogen (N) availability affects plant carbon (C) utilization. However, it is unclear how various tree functional types respond to N addition in terms of C assimilation, allocation, and storage. Here, a microcosm experiment with dual 13C and 15N labeling was conducted to study the effects of N addition (i.e., control, 0 g N kg-1; moderate N addition, 1.68 g N kg-1; and high N addition, 3.36 g N kg-1 soil) on morphological traits, on changes in nonstructural carbohydrates (NSC) in different organs, as well as on C and N uptake and allocation in three European temperate forest tree species (i.e., Acer pseudoplatanus, Picea abies and Abies alba). Our results demonstrated that root N uptake rates of the three tree species increased by N addition. In A. pseudoplatanus, N uptake by roots, N allocation to aboveground organs, and aboveground biomass allocation significantly improved by moderate and high N addition. In A. alba, only the high N addition treatment considerably raised aboveground N and C allocation. In contrast, biomass as well as C and N allocation between above and belowground tissues were not altered by N addition in P. abies. Meanwhile, NSC content as well as C and N coupling (represented by the ratio of relative 13C and 15N allocation rates in organs) were affected by N addition in A. pseudoplantanus and P. abies but not in A. alba. Overall, A. pseudoplatanus displayed the highest sensitivity to N addition and the highest N requirement among the three species, while P. abies had a lower N demand than A. alba. Our findings highlight that the responses of C and N allocation to soil N availability are species-specific and vary with the amount of N addition.

Do trees respond to pollution? A network study of the impact of pollution on spruce growth from Europe.

Tree rings have been reliably used as an environmental proxy over the past decades for environmental reconstructions, simulations and forecasting. In our study, we investigated whether tree-ring chronologies are impacted by pollution. We chose sites in the Krusné hory and the Krkonose Mountains in the Czech Republic which have a known history of pollution. We sampled Norway spruce (Picea abies [L.] Karst) in both ranges and compared their chronologies. We found no significant difference in the overall radial growth in the chronologies from both regions. However, we observed an increased heterogeneity in the growth of trees from the 1970s till the 1990s. Coherently, a severe reduction in tree growth from the late 1970s and a recovery towards the early 1990s was evident. We collected and analysed soil samples for pH and exchangeable element concentrations. All seven sampling sites' soils were strongly acidic (pHCaCl2 = 3.3 ± 0.4). The average soil base saturation at Krusné hory was higher than at Krkonose (39% versus 12%), likely due to more intensive liming. Further, we compared these chronologies to other sites in Europe. Analysing 89 sites, we found that most (9 out of 14) of the sites with significantly reduced radial tree growth were located within the former 'Black Triangle', an area which was subjected to heavy industrialisation and pollution from the 1960s to the 1990s. Atmospheric sulphur deposition was found to negatively affect radial tree-growth, while limited quantities of oxidised nitrogen appeared to have a positive effect. Our results are consistent with previous research, indicating that atmospheric SO2 pollution and subsequent acid fog and rime have led to a reduction in annual radial tree growth across the Black Triangle.

Dry and wet periods determine stem and soil greenhouse gas fluxes in a northern drained peatland forest.

Greenhouse gas (GHG) fluxes from peatland soils are relatively well studied, whereas tree stem fluxes have received far less attention. Simultaneous year-long measurements of soil and tree stem GHG fluxes in northern peatland forests are scarce, as previous studies have primarily focused on the growing season. We determined the seasonal dynamics of tree stem and soil CH4, N2O and CO2 fluxes in a hemiboreal drained peatland forest. Gas samples for flux calculations were manually collected from chambers at different heights on Downy Birch (Betula pubescens) and Norway Spruce (Picea abies) trees (November 2020-December 2021) and analysed using gas chromatography. Environmental parameters were measured simultaneously with fluxes and xylem sap flow was recorded during the growing season. Birch stems played a greater role in the annual GHG dynamics than spruce stems. Birch stems were net annual CH4, N2O and CO2 sources, while spruce stems constituted a CH4 and CO2 source but a N2O sink. Soil was a net CO2 and N2O source, but a sink of CH4. Temporal dynamics of stem CH4 and N2O fluxes were driven by isolated emissions' peaks that contributed significantly to net annual fluxes. Stem CO2 efflux followed a seasonal trend coinciding with tree growth phenology. Stem CH4 dynamics were significantly affected by the changes between wetter and drier periods, while N2O was more influenced by short-term changes in soil hydrologic conditions. We showed that CH4 emitted from tree stems during the wetter period can offset nearly half of the soil sink capacity. We presented for the first time the relationship between tree stem GHG fluxes and sap flow in a peatland forest. The net CH4 flux was likely an aggregate of soil-derived and stem-produced CH4. A dominating soil source was more evident for stem N2O fluxes.

Hybridization mediated range expansion and climate change resilience in two keystone tree species of boreal forests.

Current global climate change is expected to affect biodiversity negatively at all scales leading to mass biodiversity loss. Many studies have shown that the distribution of allele frequencies across a species' range is often influenced by specific genetic loci associated with local environmental variables. This association reflects local adaptation and allele changes at those loci could thereby contribute to the evolutionary response to climate change. However, predicting how species will adapt to climate change from this type of data alone remains challenging. In the present study, we combined exome capture sequences and environmental niche reconstruction, to test multiple methods for assessing local adaptation and climate resilience in two widely distributed conifers, Norway spruce and Siberian spruce. Both species are keystone species of the boreal forest and share a vast hybrid zone. We show that local adaptation in conifers can be detected through allele frequency variation, population-level ecological preferences, and historical niche movement. Moreover, we integrated genetic and ecological information into genetic offset predictive models to show that hybridization plays a central role in expanding the niche breadth of the two conifer species and may help both species to cope better with future changing climates. This joint genetic and ecological analysis also identified spruce populations that are at risk under current climate change.

Impacts of winter warming events on spruce budworm: the importance of timing.

Winter climate change constitutes not only a shift in chronic conditions (i.e., shorter length and warmer average temperatures) but will also influence the dynamics of extreme warming events. The latter may be particularly important for the performance and survival of insects, given their susceptibility to temperature variation. However, metabolic sensitivity changes over the course of winter diapause, and thus, insect responses to warming may vary depending on when the event occurs. To determine the influence of warm-up timing, we exposed the spruce budworm, Choristoneura fumiferana ((Clem.), Lepidoptera: Tortricidae), to acute warming events in early-, mid-, and late-dormancy and measured impacts on survival, performance, and biochemistry. While we did not observe any impacts of warm-up timing on performance, survival to the adult stage was significantly reduced in response to earlier warming. Additionally, glycogen concentration was significantly higher in response to early and late warming exposure. Collectively, these results suggest that the timing of extreme winter warming events matters, with consequences for both lethal and sublethal responses.

Complementary roles of two classes of defense chemicals in white spruce against spruce budworm.

MAIN CONCLUSION: Monoterpenes and phenolics play distinct roles in defending white spruce trees from insect defoliators. Monoterpenes contribute to the toxicity of the foliage, deterring herbivory, whereas phenolics impede budworm growth. This study demonstrates the complex interplay between monoterpenes and phenolics and their collective influence on the defense strategy of white spruce trees against a common insect defoliator. Long-lived coniferous trees display considerable variations in their defensive chemistry. The impact of these defense phenotype variations on insect herbivores of the same species remains to be thoroughly studied, mainly due to challenges in replicating the comprehensive defense profiles of trees under controlled conditions. This study methodically examined the defensive properties of foliar monoterpenes and phenolics across 80 distinct white spruce families. These families were subsequently grouped into two chemotypes based on their foliar monoterpene concentrations. To understand the separate and combined effects of these classes on tree defenses to the eastern spruce budworm, we conducted feeding experiments using actual defense profiles from representative families. Specifically, we assessed budworm response when exposed to substrates amended with phenolics alone or monoterpenes. Our findings indicate that the ratios and amounts of monoterpenes and phenolics present in the white spruce foliage influence the survival of spruce budworms. Phenotypes associated with complete larval mortality exhibited elevated ratios (ranging from 0.4 to 0.6) and concentrations (ranging from 1143 to 1796 ng mg-1) of monoterpenes. Conversely, families characterized by higher phenolic ratios (ranging from 0.62 to 0.77) and lower monoterpene concentrations (ranging from 419 to 985 ng mg-1) were less lethal to the spruce budworm. Both classes of defense compounds contribute significantly to the overall defensive capabilities of white spruce trees. Monoterpenes appear critical in determining the general toxicity of foliage, while phenolics play a role in slowing budworm development, thereby underscoring their collective importance in white spruce defenses.

Nutrient allocation patterns of Picea crassifolia on the eastern margin of the Qinghai-Tibet Plateau.

It can provide a basis for decision making for the conservation and sustainable use of forest ecosystems in mountains to understand the stoichiometric properties and nutrient allocation strategies of major tree species. However, the plant nutrient allocation strategies under different environmental gradients in forest systems of arid and semi-arid mountains are not fully understand. Therefore, three typical regions in the Qilian Mountains on the eastern edge of the Qinghai-Tibet Plateau were selected based on precipitation and temperature gradients, and the stoichiometric characteristics and nutrient allocation strategies of Qinghai spruce (Picea crassifolia) of the dominant tree species under different environmental gradients were investigated. The results showed that (1) the stoichiometric characteristics of plant tissues were different in the three regions. (2) The importance of each tissue in the plant nutrient allocation varied in different regions, showing that the plant roots are more important in the warm-wet region, while the plant leaves, branches and trunks are more important in the transition and hot-dry regions. (3) The influencing factors affecting plant nutrient allocation strategies were inconsistent across regions, which showed that plant nutrient allocation strategies in the warm-wet and transition region were mainly influenced by soil factors, while they were more influenced by climatic factors in the hot-dry region. The patterns of plant nutrient allocation strategies and drivers under different environmental gradients could help us better understand the ecological adaptation mechanism and physiological adjustment mechanism of forest ecosystem in mountains.

Drought increases Norway spruce susceptibility to the Eurasian spruce bark beetle and its associated fungi.

Drought affects the complex interactions between Norway spruce, the bark beetle Ips typographus and associated microorganisms. We investigated the interplay of tree water status, defense and carbohydrate reserves with the incidence of bark beetle attack and infection of associated fungi in mature spruce trees. We installed roofs to induce a 2-yr moderate drought in a managed spruce stand to examine a maximum of 10 roof and 10 control trees for resin flow (RF), predawn twig water potentials, terpene, phenolic and carbohydrate bark concentrations, and bark beetle borings in field bioassays before and after inoculation with Endoconidiophora polonica and Grosmannia penicillata. Drought-stressed trees showed more attacks and significantly longer fungal lesions than controls, but maintained terpene resin defenses at predrought levels. Reduced RF and lower mono- and diterpene, but not phenolic concentrations were linked with increased host selection. Bark beetle attack and fungi stimulated chemical defenses, yet G. penicillata reduced phenolic and carbohydrate contents. Chemical defenses did not decrease under mild, prolonged drought in our simulated small-scale biotic infestations. However, during natural mass attacks, reductions in carbon fixation under drought, in combination with fungal consumption of carbohydrates, may deplete tree defenses and facilitate colonization by I. typographus.

[Effects of thinning on accumulation of soil microbial residue carbon of Picea asperata plantations in sub-alpine region of western Sichuan, China]. / 间伐对川西亚高山云杉人工林土壤微生物残体碳积累的影响.

Microbial residues are an important component of soil organic carbon (SOC). It is unclear how long-term thinning affects the accumulation characteristics of microbial residue carbon (C). We analyzed the differences in soil physicochemical properties, microbial communities, extracellular enzyme activities, and microbial residue C in topsoil (0-10 cm) and subsoil (20-30 cm) in Picea asperata plantation of non-thinned (control, 4950 trees·hm-2) and thinned for 14 years (1160 trees·hm-2) stands, aiming to reveal the regulatory mechanism of thinning on microbial residue C accumulation. The results showed that thinning significantly increased SOC content, total nitrogen content, available phosphorus content, the proportion of particulate organic C, soil water content, C-cycle hydrolase, and acid phosphatase activities, but significantly reduced the proportion of mineral-associated organic C. Thinning significantly affected the content of fungal and microbial residue C, and the contribution of microbial residue C to SOC, and these effects were independent of soil layer. The content of fungal and microbial residue C was 25.0% and 24.5% higher under thinning treatments. However, thinning significantly decreased the contribution of microbial residue C to SOC by 12.3%, indicating an increase in the proportion of plant-derived C in SOC. Stepwise regression analysis showed that total nitrogen and soil water content were key factors influencing fungal and micro-bial residue C accumulation. In summary, thinning promoted microbial residue C sequestration by altering soil pro-perties and changed the composition of SOC sources.

Positive effects of tree species diversity on productivity switch to negative after severe drought mortality in a temperate forest experiment.

The synthesis of a large body of evidence from field experiments suggests more diverse plant communities are more productive as well as more resistant to the effects of climatic extremes like drought. However, this view is strongly based on data from grasslands due to the limited empirical evidence from tree diversity experiments. Here we report on the relationship between tree diversity and productivity over 10 years in a field experiment established in 2005 that was then affected by the 2018 mega-drought in central Europe. Across a number of years, tree species diversity and productivity were significantly positively related; however, the slope switched to negative in the year of the drought. Net diversity effects increased through time, with complementarity effects making greater contributions to the net diversity effect than selection effects. Complementarity effects were clearly positive in three- and five-species mixtures before the drought (2012-2016) but were found to decrease in the year of the drought. Selection effects were clearly positive in 2016 and remained positive in the drought year 2018 in two-, three-, and five-species mixtures. The survival of Norway spruce (Picea abies) plummeted in response to the drought, and a negative relationship between species diversity and spruce survival was found. Taken together, our findings suggest that tree diversity per se may not buffer communities against the impacts of extreme drought and that tree species composition and the drought tolerance of tree species (i.e., species identity) will be important determinants of community productivity as the prevalence of drought increases.

High-density genetic linkage mapping in Sitka spruce advances the integration of genomic resources in conifers.

In species with large and complex genomes such as conifers, dense linkage maps are a useful resource for supporting genome assembly and laying the genomic groundwork at the structural, populational, and functional levels. However, most of the 600+ extant conifer species still lack extensive genotyping resources, which hampers the development of high-density linkage maps. In this study, we developed a linkage map relying on 21,570 single nucleotide polymorphism (SNP) markers in Sitka spruce (Picea sitchensis [Bong.] Carr.), a long-lived conifer from western North America that is widely planted for productive forestry in the British Isles. We used a single-step mapping approach to efficiently combine RAD-seq and genotyping array SNP data for 528 individuals from 2 full-sib families. As expected for spruce taxa, the saturated map contained 12 linkages groups with a total length of 2,142 cM. The positioning of 5,414 unique gene coding sequences allowed us to compare our map with that of other Pinaceae species, which provided evidence for high levels of synteny and gene order conservation in this family. We then developed an integrated map for P. sitchensis and Picea glauca based on 27,052 markers and 11,609 gene sequences. Altogether, these 2 linkage maps, the accompanying catalog of 286,159 SNPs and the genotyping chip developed, herein, open new perspectives for a variety of fundamental and more applied research objectives, such as for the improvement of spruce genome assemblies, or for marker-assisted sustainable management of genetic resources in Sitka spruce and related species.

Structure of KOH-soluble polysaccharides from сoniferous greens of Norway spruce (Picea abies): The pectin-xylan-AGPs complex. Part 1.

Using 7 % KOH, the polysaccharide PAK has been isolated from the coniferous greens of Norway spruce. PAK was found to contain predominantly arabinoglucuronoxylan, xyloglucan and arabinan, but also pectic polysaccharides, glucomannan and arabinogalactan proteins (AGPs), as determined by 1D/2D NMR analysis. It was found that fractionation of PAK on DEAE-cellulose resulted in simultaneous elution of pectins, arabinoglucuronoxylans and AGPs. It was evident that the content of 4-OMe-α-D-GlcpA and xylose, 1,4-ß-D-GlcpA, and T-ß-D-GlcpA increased with an increase in NaCl concentration. However, 1,4-α-D-GalpA content was almost independent of NaCl concentration, indicating unchanged pectic polysaccharide concentration. Interestingly, pectins extracted with 0.1-0.3 M NaCl solutions were richer in rhamnogalacturonan-I (RG-I) than those extracted with water and 0.01 M NaCl. Conclusion: The content of RG-I, AGPs and arabinoglucuronoxylan rises with rising NaCl concentration. An intense signal indicating an intermolecular linkage between the xylan and RG-I domains, i.e. that part of the arabinoglucuronoxylan is covalently bound to RG-I, is observed in the HMBC spectra of the polysaccharides obtained. The discovery here of a new relationship between rhamnogalacturonan I and xylan contradicts the prevailing cell wall model.

Response of tree growth to drought variability in arid areas: Local hydroclimate and large-scale precipitation.

The impact of drought on terrestrial ecosystems is increasing, and the spatiotemporal heterogeneity of drought changes exacerbates the difficulty of determining ecosystem responses, especially in arid regions far from oceans. Tree rings have been widely used to understand how forest ecosystems respond to drought. However, the link between local hydroclimate variations related to tree rings and large-scale climate changes is not clear in the Qilian Mountains. Here, we used the tree ring width index to analyze the trend of Picea crassifolia growth and its relationship with climate in the middle Qilian Mountains. The results showed that the radial growth trend of Picea crassifolia is synchronized in the middle Qilian Mountains by calculating the Gleichläufigkeit index (GLK). Our analyses indicated that tree radial growth is positively correlated with drought during the growing season. Tree growth responds stably to drought (scPDSI and SPEI) and precipitation but unstably to temperature during 1950-2019. We further traced the meteorological factors that cause regional drought changes associated with radial growth. An increased total precipitation and decreased evaporation contribute to drought alleviation, favoring an increased tree radial growth. The increased total precipitation is mainly due to increased large-scale precipitation, which is related to water vapor transport changes. This study attempts to explore the influence of large-scale meteorology on regional drought change and its related tree radial growth response, which helps us to better understand the changes in forest ecosystems under climate change.

Spatial and remote sensing monitoring shows the end of the bark beetle outbreak on Belgian and north-eastern France Norway spruce (Picea abies) stands.

In 2022, Europe emerged from eight of the hottest years on record, leading to significant spruce mortality across Europe. The particularly dry weather conditions of 2018 triggered an outbreak of bark beetles (Ips typographus), causing the loss of thousands of hectares of Norway spruce stands, including in Wallonia and North-eastern France. A methodology for detecting the health status of spruce was developed based on a dense time series of satellite imagery (Sentinel-2). The time series of satellite images allowed the modelling of the spectral response of healthy spruce forests over the seasons: a decrease in photosynthetic activity of the forest canopy causes deviations from this normal seasonal vegetation index trajectory. These anomalies are caused by a bark beetle attack and are detected automatically. The method leads in the production of an annual spruce health map of Wallonia and Grand-Est. The goal of this paper is to assess the damage caused by bark beetle using the resulting spruce health maps. A second objective was to compare the influence of basic variables on the mortality of spruce trees in these two regions. Lasted 6 years (2017-2022), bark beetle has destroyed 12.2% (23,674 ha) of the spruce area in Wallonia and Grand-Est of France. This study area is composed of three bioclimatic areas: Plains, Ardennes and Vosges, which have not been equally affected by bark beetle attacks. The plains were the most affected, with 50% of spruce forests destroyed, followed by the Ardennes, which lost 11.3% of its spruce stands. The Vosges was the least affected bioclimatic area, with 5.6% of spruce stands lost. For the most problematic sites, Norway spruce forestry should no longer be considered.

Contrasting physiological strategies explain heterogeneous responses to severe drought conditions within local populations of a widespread conifer.

Understanding how trees prioritize carbon gain at the cost of drought vulnerability under severe drought conditions is crucial for predicting which genetic groups and individuals will be resilient to future climate conditions. In this study, we investigated variations in growth, tree-ring anatomy as well as carbon and oxygen isotope ratios to assess the sensitivity and the xylem formation process in response to an episode of severe drought in 29 mature white spruce (Picea glauca [Moench] Voss) families grown in a common garden trial. During the drought episode, the majority of families displayed decreased growth and exhibited either sustained or increased intrinsic water-use efficiency (iWUE), which was largely influenced by reduced stomatal conductance as revealed by the dual carbon­oxygen isotope approach. Different water-use strategies were detected within white spruce populations in response to drought conditions. Our results revealed intraspecific variation in the prevailing physiological mechanisms underlying drought response within and among populations of Picea glauca. The presence of different genetic groups reflecting diverse water-use strategies within this largely-distributed conifer is likely to lessen the negative effects of drought and decrease the overall forest ecosystems' sensitivity to it.

Picea wilsonii NAC31 and DREB2A Cooperatively Activate ERD1 to Modulate Drought Resistance in Transgenic Arabidopsis.

The NAC family of transcription factors (TFs) regulate plant development and abiotic stress. However, the specific function and response mechanism of NAC TFs that increase drought resistance in Picea wilsonii remain largely unknown. In this study, we functionally characterized a member of the PwNAC family known as PwNAC31. PwNAC31 is a nuclear-localized protein with transcriptional activation activity and contains an NAC domain that shows extensive homology with ANAC072 in Arabidopsis. The expression level of PwNAC31 is significantly upregulated under drought and ABA treatments. The heterologous expression of PwNAC31 in atnac072 Arabidopsis mutants enhances the seed vigor and germination rates and restores the hypersensitive phenotype of atnac072 under drought stress, accompanied by the up-regulated expression of drought-responsive genes such as DREB2A (DEHYDRATION-RESPONSIVE ELEMENT BINDING PROTEIN 2A) and ERD1 (EARLY RESPONSIVE TO DEHYDRATION STRESS 1). Yeast two-hybrid and bimolecular fluorescence complementation assays confirmed that PwNAC31 interacts with DREB2A and ABF3 (ABSCISIC ACID-RESPONSIVE ELEMENT-BINDING FACTOR 3). Yeast one-hybrid and dual-luciferase assays showed that PwNAC31, together with its interaction protein DREB2A, directly regulated the expression of ERD1 by binding to the DRE element of the ERD1 promoter. Collectively, our study provides evidence that PwNAC31 activates ERD1 by interacting with DREB2A to enhance drought tolerance in transgenic Arabidopsis.