Woody Plants Research Articles

Transcriptome analysis and functional validation reveal the novel role of LhCYCL in axillary bud development in hybrid Liriodendron.

Shoot branching significantly influences yield and timber quality in woody plants, with hybrid Liriodendron being particularly valuable due to its rapid growth. However, understanding of the mechanisms governing shoot branching in hybrid Liriodendron remains limited. In this study, we systematically examined axillary bud development using morphological and anatomical approaches and selected four distinct developmental stages for an extensive transcriptome analysis. A total of 9,449 differentially expressed genes have been identified, many of which are involved in plant hormone signal transduction pathways. Additionally, we identified several transcription factors downregulated during early axillarybud development, including a noteworthy gene annotated as CYC-like from the TCP TF family, which emerged as a strong candidate for modulating axillary bud development. Quantitative real-time polymerase chain reaction results confirmed the highest expression levels of LhCYCL in hybrid Liriodendron axillary buds, while histochemical β-glucuronidase staining suggested its potential role in Arabidopsis thaliana leaf axil development. Ectopic expression of LhCYCL in A. thaliana led to an increase of branches and a decrease of plant height, accompanied by altered expression of genes involved in the plant hormone signaling pathways. This indicates the involvement of LhCYCL in regulating shoot branching through plant hormone signaling pathways. In summary, our results emphasize the pivotal role played by LhCYCL in shoot branching, offering insights into the function of the CYC-like gene and establishing a robust foundation for further investigations into the molecular mechanisms governing axillary bud development in hybrid Liriodendron.

Influence of the leaf-cutting ant Atta sexdens Linnaeus (1758) on the taxonomic and functional diversity of woody plants in Atlantic Forest fragments

Influence of the leaf-cutting ant Atta sexdens Linnaeus (1758) on the taxonomic and functional diversity of woody plants in Atlantic Forest fragments

Comprehensive evaluation and application of woody plants in the green spaces of parks in saline-Alkaline areas from a low-carbon perspective: A case study of Tianjin Qiaoyuan Park.

The field of landscape architecture has placed significant emphasis on low-carbon landscapes due to the increasing challenges posed by global warming and environmental deterioration in recent years. The soil ecological conditions in saline-alkaline areas are characterized by poor quality, resulting in suboptimal growth conditions for trees. This, in turn, hampers their ability to effectively sequester carbon, thereby diminishing the potential benefits of carbon sinks. Additionally, the maintenance of tree landscapes in such areas generates more carbon emissions than does conventional green land, making it difficult to reap the benefits of tree-based carbon. A comprehensive evaluation of trees in green park spaces in saline-alkaline areas is conducted from a low-carbon perspective; by identifying the dominant tree species that are well suited to greening, we can offer a precise scientific foundation for implementing low-carbon greening initiatives in cities situated in saline-alkaline environments. Therefore, as a case study, this study investigates Tianjin Qiaoyuan Park, a typical saline park in the Bohai Bay region. The hierarchical analysis method (AHP) was used to evaluate 50 species of trees and shrubs in the park from a low-carbon perspective. The results show that the evaluation system consists of four criterion layers and 15 indicator factors. The relative weight of the criterion layer followed the order of habitat adaptability (B2) > carbon sequestration capacity (B1) > low-carbon management and conservation (B3) > landscape aesthetics (B4). The indicator layer assigned greater weight values to net assimilation (C1), saline and alkaline adaptability (C3), drought tolerance (C4), irr igation and fertilization needs (C8), growth rate (C2), and adaptability to barrenness (C5). The trees were classified into five distinct categories, with each exhibiting significant variation in terms of the strengths and weaknesses of the indicators. According to the comprehensive score, the trees were categorized into three levels. The Grade I plants exhibited the best carbon efficiency performance, comprising a total of 12 species (e.g. Sabina chinensis, Fraxinus chinensis 'Aurea' and Hibiscus syriacu), and demonstrated superior performance in all aspects. Grade II trees, consisting of 26 species (e.g Pinus tabuliformis, Paulownia fortunei, Ligustrum × vicaryi), had the second-highest comprehensive score. Moreover, Grade III trees, encompassing 12 species (e.g Acer mono, Cedrus deodara, Magnolia denudata), exhibited lower comprehensive scores. The extensive use of Grade I and II tree species is recommended in the implementation of low-carbon greening projects in the Bohai Bay region, while Grade III tree species should be judiciously utilized. The findings of this research can serve as a valuable resource for the scientific identification of tree species that are suitable for urban park green spaces in the Bohai Bay region, which is characterized by predominantly saline and alkaline soil. Additionally, the development of an evaluation system can guide the selection of low-carbon tree species when evaluating other types of saline and alkaline lands.

Genome-Wide Analysis of C/S1-bZIP Subfamilies in Populus tomentosa and Unraveling the Role of PtobZIP55/21 in Response to Low Energy

C/S1 basic leucine zipper (bZIP) transcription factors are essential for plant survival under energy deficiency. However, studies on the responses of C/S1-bZIPs to low energy in woody plants have not yet been reported. In this study, members of C/S1-bZIP subfamilies in Populus tomentosa were systematically analyzed using bioinformatic approaches. Four C-bZIPs and 10 S1-bZIPs were identified, and their protein properties, phylogenetic relationships, gene structures, conserved motifs, and uORFs were systematically investigated. In yeast two-hybrid assays, direct physical interactions between C-bZIP and S1-bZIP members were observed, highlighting their potential functional synergy. Moreover, expression profile analyses revealed that low energy induced transcription levels of most C/S1-bZIP members, with bZIP55 and bZIP21 (a homolog of bZIP55) exhibiting particularly significant upregulation. When the expression of bZIP55 and bZIP21 was co-suppressed using artificial microRNA mediated gene silencing in transgenic poplars, root growth was promoted. Further analyses revealed that bZIP55/21 negatively regulated the root development of P. tomentosa in response to low energy. These findings provide insights into the molecular mechanisms by which C/S1-bZIPs regulate poplar growth and development in response to energy deprivation.

The use of Groasis Waterboxx devices to improve mineland reclamation efforts by increasing transplant survival and growth of five native woody species

The reclamation of disturbed lands worldwide is a difficult and increasing problem. In the Mountain West region of the United States, mining activities commonly occur in zones dominated by native woody plants. The restoration of woody plants in these areas is challenging, and direct seeding efforts result in poor seedling emergence and establishment. Therefore, seedling transplants are often used, but they frequently experience high mortality during the first year, particularly due to summer stresses. We evaluated if the Groasis Waterboxx device could be used to mitigate drought stress on seedling transplants and improve their establishment on a reclaimed waste rock dump. The Waterboxx device collects precipitation into a polypropylene reservoir and slowly dispenses water onto the soil next to the seedling. To test such a hypothesis, 400 seedlings from five native woody plants (Atriplex canescens, Cercocarpus ledifolius, Pinus edulis, Purshia tridentata, and Rhus glabra) were planted with and without a Waterboxx. The Waterboxx device improved soil water potential, at 40 cm, it took 1 month longer to go below −1 MPa under a Waterboxx. Survival increased 30–65% in four of the five species, and seedling vigor increased for three of the five species. Vigor was higher with all plant species at both planting times and was significantly better in 50% of comparisons. Results support the use of the Waterboxx for improving seedling establishment on mineland overburden sites in the semiarid mountain west. The effectiveness of the Waterboxx device provides merit for future testing where drought challenges seedling establishment.

Genome-wide analysis of the poplar homeobox gene family and the identification of PtHB180 as an important regulator of plant developmental processes and drought stress response

Homeobox (HB) transcription factors have been reported to regulate various developmental processes and hormone signaling and mediate plant adaption to abiotic stresses, including drought. As a major subfamily of HB family, KNOX genes have been widely studied in herbaceous plants. However, the behaviors of KNOX genes in regulating plant developmental processes and stress resistance in woody plants remain poorly understood. Here, we found a KNOX family gene in the model tree species Populus trichocarpa (hereafter “poplar”), PtHB180, which enhances drought tolerance by regulating developmental processes through auxin and gibberellin pathways and mediating the ABA signaling pathway leading to rapid stomatal closure. Overexpression lines of PtHB180 (PtHB180-OE) displayed pleiotropic phenotypes, including increased plant height and decreased leaf area by affecting auxin and/or gibberellin contents and signaling. PtHB180-OE plants also displayed improved drought tolerance under short- and long-term drought stress. Results of transcriptome sequencing suggested the involvement of genes related to ABA signaling in regulating drought tolerance in PtHB180-OE. Specifically, PtHB180-OE plants showed significantly enhanced stomatal sensitivity to exogenous ABA. Taken together, our results indicate that PtHB180 enhances drought tolerance in poplar via regulating plant architecture and mediating ABA signaling to close stomata. Therefore, PtHB180 can be used as a potential candidate gene to improve drought tolerance by genetic transformation in poplar and related plant species.

Genome-wide identification of WRKY transcription factors in Casuarina equisetifolia and the function analysis of CeqWRKY11 in response to NaCl/NaHCO3 stresses

BackgroundCasuarina equisetifolia (C. equisetifolia) is a woody species with many excellent features. It has natural resistance against drought, salt and saline-alkali stresses. WRKY transcription factors (TFs) play significant roles in plant response to abiotic stresses, therefore, molecular characterization of WRKY gene family under abiotic stresses holds great significance for improvement of forest trees through molecular biological tools. At present, WRKY TFs from C. equisetifolia have not been thoroughly studied with respect to their role in salt and saline-alkali stresses response. The current study was conducted to bridge the same knowledge gap.ResultsA total of 64 WRKYs were identified in C. equisetifolia and divided into three major groups i.e. group I, II and III, consisting of 10, 42 and 12 WRKY members, respectively. The WRKY members in group II were further divided into 5 subgroups according to their homology with Arabidopsis counterparts. WRKYs belonging to the same group exhibited higher similarities in gene structure and the presence of conserved motifs. Promoter analysis data showed the presence of various response elements, especially those related to hormone signaling and abiotic stresses, such as ABRE (ABA), TGACG (MeJA), W-box ((C/T) TGAC (T/C)) and TC-rich motif. Tissue specific expression data showed that CeqWRKYs were mainly expressed in root under normal growth conditions. Furthermore, most of the CeqWRKYs were up-regulated by NaCl and NaHCO3 stresses with few of WRKYs showing early responsiveness to both stresses while few others exhibiting late response. Although the expressions of CeqWRKYs were also induced by cold stress, the response was delayed compared with other stresses. Transgenic C. equisetifolia plants overexpressing CeqWRKY11 displayed lower electrolyte leakage, higher chlorophyll content, and enhanced tolerance to both stresses. The higher expression of abiotic stress related genes, especially CeqHKT1 and CeqPOD7, in overexpression lines points to the maintenance of optimum Na+/K+ ratio, and ROS scavenging as possible key molecular mechanisms underlying salt stress tolerance.ConclusionsOur results show that CeqWRKYs might be key regulators of NaCl and NaHCO3 stresses response in C. equisetifolia. In addition, positive correlation of CeqWRKY11 expression with increased stress tolerance in C. equisetifolia encourages further research on other WRKY family members through functional genomic tools. The best candidates could be incorporated in other woody plant species for improving stress tolerance.

Liebig review: The role of mineral nutrients in the development of Pseudomonas syringae diseases—Lessons learned and implications for disease control in woody plants

AbstractBackgroundThe plant pathogen Pseudomonas syringae (Ps) causes diseases in a broad range of hosts including important cash crops from several climate zones. In particular for woody crops, effective and environmentally friendly disease control strategies are not available. Although there is increasing evidence of a link between plant nutritional status and disease susceptibility, optimization of plant nutrition is often neglected in efforts to control Ps diseases. This review brings current knowledge on this topic together with the aim of facilitating the development of treatment recommendations in specific contexts.Literature reviewThe article consists of three parts. First, we compiled data on the impact of mineral nutrients on Ps disease severity in woody and herbaceous species. Next, we discuss how nutrients may be related to plant defense and/or Ps virulence. Last, we consider these findings in the context of woody hosts and give suggestions for future research.ConclusionsWe encourage a research focus on typical nutrient imbalances (deficiencies and surpluses) in specific orchard regions; testing the role of foliar fertilizers in spring (the period of highest infection risk); analyses of the interaction between nutrient supply and the microbiome in the phyllosphere; investigating the interaction between nutrient supply and other control measures, and the impact of nutrient supply on Ps diseases in the presence of other stress factors.

Multi-elements linear discriminant analysis of herbaceous and woody plants in southwest china karst region using orthogonal partial least squares model

Multi-elements linear discriminant analysis of herbaceous and woody plants in southwest china karst region using orthogonal partial least squares model

Production of Polyphenolic Natural Products by Bract-Derived Tissue Cultures of Three Medicinal Tilia spp.: A Comparative Untargeted Metabolomics Study

Medicinal plant tissue cultures are potential sources of bioactive compounds. In this study, we report the chemical characterization of the callus cultures of three medicinal Tilia spp. (Tilia cordata, Tilia vulgaris and Tilia tomentosa), along with the comparison to bracts and flowers of the same species. Our aim was to show that calli of Tilia spp. are good alternatives to the calli of T. americana for the production of polyphenols and are better sources of a subset of polyphenolic metabolites, compared to the original organs. Calli were initiated from young bracts and grown on woody plant medium containing 1 mg L−1 2,4-D and 0.1 mg L−1 BAP. For chemical characterization, a quality-controlled untargeted metabolomics approach and the quantification of several bioactive compounds was performed with the use of LC-ESI-MS/MS. While bracts and flowers contained flavonoid glycosides (astragalin, isoquercitrin) as major polyphenols, calli of all species contained catechins, coumarins (fraxin, esculin and scopoletin) and flavane aglyca. T. tomentosa calli contained 5397 µg g DW−1 catechin, 201 µg g DW−1 esculin, 218 µg g DW−1 taxifolin and 273 µg g DW−1 eriodictyol, while calli from other species contained lower amounts. T. cordata and T. tomentosa flowers were rich in isoquercitrin, containing 8134 and 6385 µg g DW−1, respectively. The currently tested species contained many of the bioactive metabolites described from T. americana. The production of catechin was shown to be comparable to the most efficient tissue cultures reported. Flowers and bracts contained flavonoid glycosides, including tiliroside, resembling bioactive fractions of T. americana. In addition, untargeted metabolomics has shown fingerprint-like differences among species, highlighting possible chemotaxonomic and quality control applications, especially for bracts.

New Understanding of Meta-Topolin Riboside Metabolism in Micropropagated Woody Plants.

Topolin cytokinins have emerged as valuable tools in micropropagation. This study investigates the metabolism of meta-topolin riboside (mTR) in three distinct tree species: Handroanthus guayacan and Tabebuia rosea (Bignoniaceae), and Tectona grandis (Lamiaceae). Employing labeled N15 mTR, we unraveled the complex mechanisms underlying cytokinin homeostasis, identifying N9-glucosylation as the principal deactivation pathway. Our findings demonstrate a capacity in T. rosea and H. guayacan to reposition the hydroxyl group on the cytokinin molecule, a previously unexplored metabolic pathway. Notably, this study reveals remarkable interfamilial and interspecies differences in mTR metabolism, challenging established perspectives on the role of callus tissue in cytokinin storage. These insights not only illuminate the metabolic intricacies of mTR, a cytokinin with interesting applications in plant tissue culture, but also enhances our understanding of cytokinin dynamics in plant systems, thereby enriching the scientific discourse on plant physiology and cytokinin biology.

Kansas Prairie Streams Are Getting Choked, Maybe for Good

A herculean effort to fight back woody plants in the Konza Prairie has largely failed. The outcome shows how difficult it can be to retore these ecosystems.

A modified CTAB method for the extraction of high-quality RNA from mono-and dicotyledonous plants rich in secondary metabolites

BackgroundHigh-quality RNA extraction from woody plants is difficult because of the presence of polysaccharides and polyphenolics that bind or co-precipitate with the RNA. The CTAB (cetyl trimethylammonium bromide) based method is widely used for the isolation of nucleic acids from polysaccharide-rich plants. Despite the widespread use of the CTAB method, it is necessary to adapt it to particular plant species, tissues and organs. Here we described a simple and generalized method for RNA isolation from mature leaf tissues of several economically important woody (17) and herbaceous plants (2) rich in secondary metabolites. High yields were achieved from small amount (up to 50 mg) of plant material. Two main modifications were applied to the basic protocol: an increase in β-mercaptoethanol concentration (to 10%v/v) and the use of an effective DNase treatment. As opposed to similar studies, we tried to describe a more detailed protocol for isolating RNA, including the exact quantity and concentration of the reagents were used.ResultsOur modified CTAB method is proved to be efficient in extracting the total RNA from a broad range of woody and herbaceous species. The RNA yield was ranged from 2.37 to 91.33 µg/µl. The A260:A280 and A260:A230 absorbance ratios were measured from 1.77 to 2.13 and from 1.81 to 2.22. The RIN value (RNA Integrity Number) of the samples fell between 7.1 and 8.1, which indicated that a small degree of RNA degradation occurred during extraction. The presence of a single peak in the melt curve analyses and low standard errors of the Ct values of replicated measurements indicated the specificity of the primers to bind to the cDNA.ConclusionsOur RNA isolation method, with fine-tuned and detailed instructions, can produce high quality RNA from a small amount of starting plant material that is suitable for use in downstream transcriptional analyses. The use of an increased concentration of the reducing agent β-mercaptoethanol in the extraction buffer, as well as the application of DNaseI-treatment resulted in a method suitable for a wide range of plants without the need of further optimalization, especially in Rhus typhina (Staghorn sumac), for which molecular-genetic studies have not yet been sufficiently explored.

Net O2 exchange rates under dark and light conditions across different stem compartments.

Woody plants display some photosynthetic activity in stems, but the biological role of stem photosynthesis and the specific contributions of bark and wood to carbon uptake and oxygen evolution remain poorly understood. We aimed to elucidate the functional characteristics of chloroplasts in stems of different ages in Fraxinus ornus. Our investigation employed diverse experimental approaches, including microsensor technology to assess oxygen production rates in whole stem, bark, and wood separately. Additionally, we utilized fluorescence lifetime imaging microscopy (FLIM) to characterize the relative abundance of photosystems I and II (PSI : PSII chlorophyll ratio) in bark and wood. Our findings revealed light-induced increases in O2 production in whole stem, bark, and wood. We present the radial profile of O2 production in F. ornus stems, demonstrating the capability of stem chloroplasts to perform light-dependent electron transport. Younger stems exhibited higher light-induced O2 production and dark respiration rates than older ones. While bark emerged as the primary contributor to net O2 production under light conditions, our data underscored that wood chloroplasts are also photosynthetically active. The FLIM analysis unveiled a lower PSI abundance in wood than in bark, suggesting stem chloroplasts are not only active but also acclimate to the spectral composition of light reaching inner compartments.

Effect of artisanal gold mining on woody plant diversity in Western Burkina Faso

Although artisanal gold mining strongly contribute to enhance people incomes in West Africa, this activity can lead to habitat fragmentation and loss of woody species. Therefore, studying the impact of artisanal gold mining on woody plant diversity is needed for efficient management of forest resources. In this study, we investigated the impact of artisanal gold mining on woody plant diversity and dynamics in the Kari classified forest in Western Burkina Faso. Forest inventories were carried out at two sites including a gold panning site and a control site, following a systematic sampling design. We identified all woody species and measured all woody species with a diamtere at breast height ≥5 cm. A total 60 plots of 900 m² were surveyed. Diversity indices and dendrometric parameters were calculated and compared between the two sites. The results showed a higher species richness (59 woody species distributing in 39 genera and 18 families) in the control site compared to the gold panning site (38 species distributing in 25 genera and 12 families). The mean values of the number of species per plot and the Shannon diversity index were higher in the control site (15.50 ± 8.80; 2.16 ± 0.43, respectively) than in the artisal mining site (5.47 ± 2.71; 1.40 ± 0.53, respectively), showing that the control site is more diverse than the site that is under gold panning. The control site also had the highest density (683.33 ± 262.99 ft/ha) and stable woody stands compared to the less gold panning site (137.41 ± 102.31 ft/ha) where unstable woody stands were found. While the gold-panning site was characterised by biophysical impacts such as destruction of the plant cover and soil excavation, no visible impact was observed at the control site.Theses findings highlight the negative impact of artisanl gold mining on woody species diversity and structure. The study provides essential information for decision-making on sustainable forest management in the context of mining boom in Burkina Faso's.

Structural patterns and regeneration status of woody plant species in the forest of Northwest Ethiopia

Structural patterns and regeneration status of woody plant species in the forest of Northwest Ethiopia

Repeated clearing as a mechanism for savanna recovery following bush encroachment

Abstract Many savannas are experiencing increased cover of trees and shrubs, resulting in reduced herbaceous productivity, shifts in savanna functional structure and potential reductions in ecotourism. Clearing woody plants has been suggested as an effective management strategy to mitigate these effects and restore these systems to an open state with higher rates of grass production and herbivory. This study investigated the effectiveness of repeated shrub clearing as a tool to mitigate bush encroachment in a semi‐arid savanna in southern Africa. We present data from a 7‐year experiment in the Mthimkhulu Game Reserve bordering Kruger National Park, South Africa. Colophospermum mopane stems and resprouting shoots were basally cut 2–3 times per year (2015–2022) in three pairs of treatment and control plots of 60 × 60 m. We monitored changes in soil moisture, grass biomass and herbivore activity via dung counts. We assessed C. mopane physiological responses to repeated cutting using non‐structural carbohydrates and stable water isotopes to infer changes to energy storage and functional rooting depth, respectively. The cleared treatment had higher soil moisture and grass biomass than the control treatment. Dung counts showed impala and buffalo visited the cleared treatment more frequently than the control treatment. Repeated cutting had limited effects on C. mopane survival in the first 2–3 years after initial clearing, but 80% of individuals were dead after 7 years. Repeatedly cut C. mopane had lower belowground starch concentrations and used water from shallower soil depths than C. mopane in control plots. Synthesis and applications. Repeated cutting increased soil moisture availability and grass biomass, and attracted charismatic grazing herbivores. While more costly than once‐off clearing methods, this practice created more employment opportunities for a neighbouring rural community. Transforming portions of the ecosystem to a grass‐dominated state may increase ecotourism potential through improved game viewing in open systems.

Uptake and translocation of pharmaceutically active compounds by olive tree (Olea europaea L.) irrigated with treated municipal wastewater

IntroductionThe use of treated municipal wastewater (TWW) represents a relevant opportunity for irrigation of agricultural crops in semi-arid regions to counter the increasing water scarcity. Pharmaceutically active compounds (PhACs) are often detected in treated wastewater, posing a risk to humans and the environment. PhACs can accumulate in soils and translocate into different plant tissues, reaching, in some cases, edible organs and entering the food chain.MethodsThis study evaluated the uptake and translocation processes of 10 PhACs by olive trees irrigated with TWW, investigating their accumulation in different plant organs. The experiment was conducted in southern Italy, in 2-year-old plants irrigated with three different types of water: freshwater (FW), TWW spiked with 10 PhACs at a concentration of 200 µg L−1 (1× TWW), and at a triple dose (3× TWW), from July to October 2021. The concentration of PhACs in soil and plant organs was assessed, collecting samples of root, stem, shoot, leaf, fruit, and kernel at 0 (T0), 50 (T1), and 107 (T2) days of irrigation. PhACs extraction from soil and plant organs was carried out using the QuEChERS method, and their concentrations were determined by high-resolution mass spectrometry coupled with liquid chromatography.ResultsResults of uptake factors (UF) showed a different behavior between compounds according to their physicochemical properties, highlighting PhACs accumulation and translocation in different plant organs (also edible part) in 1× TWW and 3× TWW compared to FW. Two PhACs, carbamazepine and fluconazole, showed interactions with the soil–plant system, translocating also in the aerial part of the plant, with a translocation factor (TF) greater than 1, which indicates high root-to-leaf translocation.DiscussionFindings highlight that only few PhACs among the selected compounds can be uptaken by woody plants and accumulated in edible parts at low concentration. No effects of PhACs exposure on plant growth have been detected. Despite the attention to be paid to the few compounds that translocate into edible organs, these results are promising for adapting wastewater irrigation in crops. Increasing knowledge about PhACs behavior in woody plants can be important for developing optimized wastewater irrigation and soil management strategies to reduce PhACs accumulation and translocation in plants.

Physiological, Biochemical, and Molecular Analyses Reveal Dark Heartwood Formation Mechanism in Acacia melanoxylon.

Acacia melanoxylon is highly valued for its commercial applications, with the heartwood exhibiting a range of colors from dark to light among its various clones. The underlying mechanisms contributing to this color variation, however, have not been fully elucidated. In an effort to understand the factors that influence the development of dark heartwood, a comparative analysis was conducted on the microstructure, substance composition, differential gene expression, and metabolite profiles in the sapwood (SW), transition zone (TZ), and heartwood (HW) of two distinct clones, SR14 and SR25. A microscopic examination revealed that heartwood color variations are associated with an increased substance content within the ray parenchyma cells. A substance analysis indicated that the levels of starches, sugars, and lignin were more abundant in SP compared to HW, while the concentrations of phenols, flavonoids, and terpenoids were found to be higher in HW than in SP. Notably, the dark heartwood of the SR25 clone exhibited greater quantities of phenols and flavonoids compared to the SR14 clone, suggesting that these compounds are pivotal to the color distinction of the heartwood. An integrated analysis of transcriptome and metabolomics data uncovered a significant accumulation of sinapyl alcohol, sinapoyl aldehyde, hesperetin, 2', 3, 4, 4', 6'-peptahydroxychalcone 4'-O-glucoside, homoeriodictyol, and (2S)-liquiritigenin in the heartwood of SR25, which correlates with the up-regulated expression of CCRs (evm.TU.Chr3.1751, evm.TU.Chr4.654_667, evm.TU.Chr4.675, evm.TU.Chr4.699, and evm.TU.Chr4.704), COMTs (evm.TU.Chr13.3082, evm.TU.Chr13.3086, and evm.TU.Chr7.1411), CADs (evm.TU.Chr10.2175, evm.TU.Chr1.3453, and evm.TU.Chr8.1600), and HCTs (evm.TU.Chr4.1122, evm.TU.Chr4.1123, evm.TU.Chr8.1758, and evm.TU.Chr9.2960) in the TZ of A. melanoxylon. Furthermore, a marked differential expression of transcription factors (TFs), including MYBs, AP2/ERFs, bHLHs, bZIPs, C2H2s, and WRKYs, were observed to be closely linked to the phenols and flavonoids metabolites, highlighting the potential role of multiple TFs in regulating the biosynthesis of these metabolites and, consequently, influencing the color variation in the heartwood. This study facilitates molecular breeding for the accumulation of metabolites influencing the heartwood color in A. melanoxylon, and offers new insights into the molecular mechanisms underlying heartwood formation in woody plants.

Gymnosperms demonstrate patterns of fine‐root trait coordination consistent with the global root economics space

Abstract Gymnosperms encompass a diverse group of mostly woody plants with high ecological and economic value, yet little is known about the scope and organization of fine‐root trait diversity among gymnosperms due to the undersampling of most gymnosperm families and the dominance of angiosperm groups in recent syntheses. New and existing data were compiled for morphological traits (root diameter, length, tissue density, specific root length [SRL] and specific root area [SRA]), the architectural trait branching ratio, root nitrogen content [N] and mycorrhizal colonization. We used phylogenetic least squares regression and principal component analysis to determine trait–trait relationships and coordination across 66 species, representing 11 of the 12 extant gymnosperm families from boreal, temperate, subtropical and tropical biomes. Finally, we compared the relationship between family divergence time and mean trait values to determine whether evolutionary history structured variation in fine‐root traits within the gymnosperm phylogeny. Wide variation in gymnosperm root traits could be largely captured by two primary axes of variation defined by SRL and diameter, and root tissue density and root nitrogen, respectively. However, individual root length and SRA each had significant correlations with traits defining both main axes of variation. Neither mycorrhizal colonization nor root branching ratio were closely related to other traits. We did not observe a directional evolution of mean trait values from older to more recently diverged gymnosperm families. Synthesis. Despite their unique evolutionary history, gymnosperms display a root economic space similar to that identified in angiosperms, likely reflecting common constraints on plants adapting to diverse environments in both groups. These findings provide greater confidence that patterns observed in broad syntheses justly capture patterns of trait diversity among multiple, distinct lineages. Additionally, independence between root architecture and other traits may support greater diversity in below‐ground resource acquisition strategies. Unlike angiosperms, there were no clear trends towards increasingly thin roots over evolutionary time, possibly because of lower diversification rates or unique biogeographic history among gymnosperms, though additional observations are needed to more richly test evolutionary trends among gymnosperms.