Wood Formation Research Articles

Reflections on the contributions of Populus research at Rhinelander, Wisconsin, USA

The United States Department of Agriculture Forest Service established three regional Institutes of Forest Genetics in the United States in the 1950s to improve trees for reforestation and improve the management of forests. The institute in Rhinelander, Wisconsin, started in 1956 as part of the Lake States Forest Experiment Station. Since that time, the “Rhinelander Lab” has undergone changes in research priorities, organizational changes, and name changes while becoming an international center of forest scientific excellence. Many of the researchers’ key findings over the years were published in the Canadian Journal of Forest Research. In this paper, for the 50th anniversary edition of the Journal, we reflect upon one part of those accomplishments: the history of the contributions of Populus L. research at Rhinelander. We discuss major research programs and the scientists conducting this work, including (i) physiology of wood formation, (ii) short rotation intensive culture and short rotation woody crops, (iii) intensively cultured plantations, (iv) physiology and utilization of short rotation poplar yields, (v) breeding and selection, (vi) biotechnology and molecular genetics, (vii) atmospheric pollution and climate change, (viii) phytotechnologies, and (ix) ecosystem services. Also, we describe four major international conferences held in Rhinelander and (or) hosted by Rhinelander researchers.

Integrative analysis of wood biomass and developing xylem transcriptome provide insights into mechanisms of lignin biosynthesis in wood formation of Pinus massoniana

Lignin is an important renewable energy source as an excellent new battery fuel and ideal substitutes for the petrochemical industry. However, the molecular mechanism underlying lignin biosynthesis in wood formation of P. massoniana remains unexplored. Thus, an integrative analysis of wood biomass and the developing xylem transcriptome was performed to identify genes involved in lignin biosynthesis. A total of 1624 differentially expressed genes (DEGs) were identified, consisting of 797 upregulated and 827 downregulated genes (MaxG vs MinG). Additionally, 122 candidate genes and 17 DEGs were successfully annotated to the lignin biosynthesis pathway. All upregulated MYB and NAC genes were regulators of secondary cell wall formation. Moreover, the qRT-PCR analyses shown that 9 lignin biosynthesis-related genes and 7 transcription factor-encoding genes were upregulated (MaxG vs MinG), which indicated that the downregulation of lignin biosynthesis-related genes might be the possible causes of growth retardation and dwarf phenotype in some P. massoniana individuals. The identification of lignin biosynthesis-related genes can provide valuable genetic basis and resource for further researches on molecular mechanisms of lignin biosynthesis and contribute to the future investigations of bioengineering and synthetic biology to regulate lignin content in wood formation for the pulp and wood utilization industry.

Genome-wide analysis of lncRNA and mRNA expression and endogenous hormone regulation during tension wood formation in Catalpa bungei

BackgroundPhytohormones are the key factors regulating vascular development in plants, and they are also involved in tension wood (TW) formation. Although the theory of hormone distribution in TW formation is widely supported, the effects of endogenous hormones on TW formation have not yet been assessed. In this study, TW formation was induced in Catalpa bungei by artificial bending. The phytohormone content of TW, opposite wood (OW) and normal wood (NW) was determined using liquid chromatography-mass spectrometry (LC-MS), and transcriptome sequencing was performed. The hormone content and related gene expression data were comprehensively analyzed.ResultsThe results of analyses of the plant hormone contents indicated significantly higher levels of cis-zeatin (cZ), indoleacetic acid (IAA) and abscisic acid (ABA) in TW than in OW. Genes involved in the IAA and ABA synthesis pathways, such as ALDH (evm.model.group5.1511) and UGT (evm.model.scaffold36.20), were significantly upregulated in TW. and the expression levels of ARF (evm.model.group5.1332), A-ARR (evm.model.group0.1600), and TCH4 (evm.model.group2.745), which participate in IAA, cZ and Brassinolide (BR) signal transduction, were significantly increased in TW. In particular, ARF expression may be regulated by long noncoding RNAs (lncRNAs) and the HD-ZIP transcription factor ATHB-15.ConclusionsWe constructed a multiple hormone-mediated network of C. bungei TW formation based on hormone levels and transcriptional expression profiles were identified during TW formation.

Alternative splicing and its regulatory role in woody plants.

Alternative splicing (AS) is an important post-transcriptional process to enhance proteome diversity in eukaryotic organisms. In plants, numerous reports have primarily focused on AS analysis in model plant species or herbaceous plants, leading to a notable lack of research on AS in woody plants. More importantly, emerging evidence indicates that many important traits, including wood formation and stress resistance, in woody plants are controlled by AS. In this review article, we summarize the current progress of all kinds of AS studies in different tree species at various stages of development and in response to various stresses, revealing the significant role played by AS in woody plants, as well as the similar properties and differential regulation within their herbaceous counterparts. Furthermore, we propose several potential strategies to facilitate the functional characterization of splicing factors in woody plants and evaluate a general pipeline for the systematic characterization of splicing isoforms in a complex AS regulatory network. The utilization of genetic studies and high-throughput omics integration approaches to analyze AS genes and splicing factors is likely to further advance our understanding of AS modulation in woody plants.

A metabolite roadmap of the wood-forming tissue in Populus tremula.

Wood, or secondary xylem, is the product of xylogenesis, a developmental process that begins with the proliferation of cambial derivatives and ends with mature xylem fibers and vessels with lignified secondary cell walls. Fully mature xylem has undergone a series of cellular processes, including cell division, cell expansion, secondary wall formation, lignification and programmed cell death. A complex network of interactions between transcriptional regulators and signal transduction pathways controls wood formation. However, the role of metabolites during this developmental process has not been comprehensively characterized. To evaluate the role of metabolites during wood formation, we performed a high spatial resolution metabolomics study of the wood-forming zone of Populus tremula, including laser dissected aspen ray and fiber cells. We show that metabolites show specific patterns within the wood-forming zone, following the differentiation process from cell division to cell death. The data from profiled laser dissected aspen ray and fiber cells suggests that these two cell types host distinctly different metabolic processes. Furthermore, by integrating previously published transcriptomic and proteomic profiles generated from the same trees, we provide an integrative picture of molecular processes, for example, deamination of phenylalanine during lignification is of critical importance for nitrogen metabolism during wood formation.

Frost and drought: Effects of extreme weather events on stem carbon dynamics in a Mediterranean beech forest.

The effects of short-term extreme events on tree functioning and physiology are still rather elusive. European beech is one of the most sensitive species to late frost and water shortage. We investigated the intra-annual C dynamics in stems under such conditions. Wood formation and stem CO2 efflux were monitored in a Mediterranean beech forest for 3 years (2015-2017), including a late frost (2016) and a summer drought (2017). The late frost reduced radial growth and, consequently, the amount of carbon fixed in the stem biomass by 80%. Stem carbon dioxide efflux in 2016 was reduced by 25%, which can be attributed to the reduction of effluxes due to growth respiration. Counter to our expectations, we found no effects of the 2017 summer drought on radial growth and stem carbon efflux. The studied extreme weather events had various effects on tree growth. Even though late spring frost had a strong impact on beech radial growth in the current year, trees fully recovered in the following growing season, indicating high resilience of beech to this stressful event.

Evolutionary network genomics of wood formation in a phylogenetic survey of angiosperm forest trees.

Wood formation was present in early angiosperms, but has been highly modified through evolution to generate the anatomical diversity seen in extant angiosperm lineages. In this project, we modeled changes in gene coexpression relationships associated with the evolution of wood formation in a phylogenetic survey of 13 angiosperm tree species. Gravitropic stimulation was used as an experimental treatment to alter wood formation and also perturb gene expression. Gene transcript abundances were determined using RNA sequencing of developing wood tissues from upright trees, and from the top (tension wood) and bottom (opposite wood) tissues of gravistimulated trees. A network-based approach was employed to align gene coexpression networks across species based on orthologous relationships. A large-scale, multilayer network was modeled that identified both lineage-specific gene coexpression modules and modules conserved across multiple species. Functional annotation and analysis of modules identified specific regulatory processes associated with conserved modules, including regulation of hormones, protein phosphorylation, meristem development and epigenetic processes. Our results provide novel insights into the evolution and development of wood formation, and demonstrate the ability to identify biological processes and genes important for the evolution of a foundational trait in nonmodel, undomesticated forest trees.

Comparing the Cell Dynamics of Tree-Ring Formation Observed in Microcores and as Predicted by the Vaganov-Shashkin Model.

New insights into the intra-annual dynamics of tree-ring formation can improve our understanding of tree-growth response to environmental conditions at high-resolution time scales. Obtaining this information requires, however, a weekly monitoring of wood formation, sampling that is extremely time-intensive and scarcely feasible over vast areas. Estimating the timing of cambial and xylem differentiation by modeling thus represents an interesting alternative for obtaining this important information by other means. Temporal dynamics of cambial divisions can be extracted from the daily tree-ring growth rate computed by the Vaganov–Shashkin (VS) simulation model, assuming that cell production is tightly linked to tree-ring growth. Nonetheless, these predictions have yet to be compared with direct observations of wood development, i.e., via microcoring, over a long time span. We tested the performance of the VS model by comparing the observed and predicted timing of wood formation in black spruce [Picea mariana (Mill.)]. We obtained microcores over 15 years at 5 sites along a latitudinal gradient in Quebec (Canada). The measured variables included cell size and the timing of cell production and differentiation. We calibrated the VS model using daily temperature and precipitation recorded by weather stations located on each site. The predicted and observed timing of cambial and enlarging cells were highly correlated (R 2 = 0.8); nonetheless, we detected a systematic overestimation in the predicted timing of cambial cells, with predictions delayed by 1–20 days compared with observations. The growth rate of cell diameter was correlated with the predicted growth rate assigned to each cambial cell, confirming that cell diameter developmental dynamics have the potential to be inferred by the tree-ring growth curve of the VS model. Model performances decrease substantially in estimating the end of wood formation. The systematic errors suggest that the actual relationships implemented in the model are unable to explain the phenological events in autumn. The mismatch between the observed and predicted timing of wood formation in black spruce within our study area can be reduced by better adapting the VS model to wet sites, a context for which this model has been rarely used.

Age Dynamics of Wind Risk and Tree Sway Characteristics in a Softwood Plantation

Quantifying wind risk in planted forests is critical to ensure the provision of wood-based biomaterials and forest ecosystem services in the future. Tree sway dynamics play a major role in determining their vulnerability to wind storms. While structural engineering theory can greatly assist in predicting tree and branch motion accurately, the effects of forest aging and silvicultural treatments on motion and the likelihood of mechanical failure are still poorly understood. In this study, we simulate the growth of an individual model tree and its architecture for a managed, even-aged pure Cryptomeria japonica D. Don. stand. The dynamic behavior of the tree is predicted for a rotation period at a five-year interval using finite element analysis. Results show that tree natural sway frequency is subject to a five-fold decrease over the tree’s lifetime. The tree is thus more susceptible to absorb turbulent kinetic energy from the wind as it grows older. The damping ratio, which measures how rapidly the tree can return to its rest position after being deflected, is predicted to increase by a factor of four. By simulating the mechanical response to a wind gust, the model predicts a quick transition from a low risk of failure in the juvenile phase to a high-risk situation in the mature phase. The transition occurs between ages 15 and 30 when canopy closure takes place, when juvenile wood formation stops in the lower stem region and when the first thinning is carried out. Wind risk plateaus after the transition period. The age-risk relationship shown by the model is consistent with post-typhoon damage patterns documented by inventories in Japanese planted forests, albeit with a sharper transition.

Photoperiod and temperature as dominant environmental drivers triggering secondary growth resumption in Northern Hemisphere conifers.

Wood formation consumes around 15% of the anthropogenic CO2 emissions per year and plays a critical role in long-term sequestration of carbon on Earth. However, the exogenous factors driving wood formation onset and the underlying cellular mechanisms are still poorly understood and quantified, and this hampers an effective assessment of terrestrial forest productivity and carbon budget under global warming. Here, we used an extensive collection of unique datasets of weekly xylem tissue formation (wood formation) from 21 coniferous species across the Northern Hemisphere (latitudes 23 to 67°N) to present a quantitative demonstration that the onset of wood formation in Northern Hemisphere conifers is primarily driven by photoperiod and mean annual temperature (MAT), and only secondarily by spring forcing, winter chilling, and moisture availability. Photoperiod interacts with MAT and plays the dominant role in regulating the onset of secondary meristem growth, contrary to its as-yet-unquantified role in affecting the springtime phenology of primary meristems. The unique relationships between exogenous factors and wood formation could help to predict how forest ecosystems respond and adapt to climate warming and could provide a better understanding of the feedback occurring between vegetation and climate that is mediated by phenology. Our study quantifies the role of major environmental drivers for incorporation into state-of-the-art Earth system models (ESMs), thereby providing an improved assessment of long-term and high-resolution observations of biogeochemical cycles across terrestrial biomes.

Genome Cytosine Methylation May Affect Growth and Wood Property Traits in Populations of Populus tomentosa

Growth and wood formation are crucial and complex biological processes during tree development. These biological regulatory processes are presumed to be controlled by DNA methylation. However, there is little direct evidence to show that genes taking part in wood regulation are affected by cytosine methylation, resulting in phenotypic variations. Here, we detected epimarkers using a methylation-sensitive amplification polymorphism (MSAP) method and performed epimarker–trait association analysis on the basis of nine growth and wood property traits within populations of 432 genotypes of Populus tomentosa. Tree height was positively correlated with relative full-methylation level, and 1101 out of 2393 polymorphic epimarkers were associated with phenotypic traits, explaining 1.1–7.8% of the phenotypic variation. In total, 116 epimarkers were successfully sequenced, and 96 out of these sequences were linked to putative genes. Among them, 13 candidate genes were randomly selected for verification using quantitative real-time PCR (qRT-PCR), and it also showed the expression of nine putative genes of PtCYP450, PtCpn60, PtPME, PtSCP, PtGH, PtMYB, PtWRKY, PtSTP, and PtABC were negatively correlated with DNA methylation level. Therefore, it suggested that changes in DNA methylation might contribute to regulating tree growth and wood property traits.

Anatomical Features and Its Radial Variations among Different Catalpa bungei Clones

Research highlights: Annual wood anatomy (xylem) aids our understanding of mature wood formation and the growth strategies of trees. Background and Objectives: Catalpa bungei is an important native species in China that produces excellent quality wood. Herein, we clarified the effects of the genetic origin and cambial age on the anatomical characteristics of C. bungei wood. Materials and Methods: Six new 13-year-old C. bungei clones: ‘1-1’ (n trees = 3), ‘1-3’ (n trees = 3), ‘2-7’ (n trees = 3), ‘2-8’ (n trees = 3), ‘8-1’ (n trees = 4), and ‘9-1’ (n trees = 3) were removed for study from a plantation in Tianshui City, Gansu province, China. Xylem features were observed and the anatomical variables were manually measured via image analysis on (macro- micro-, and ultra-) features cut from radial increments of earlywood and latewood sampled at breast height. Results: Between the age of 1 and 2 years, wood was diffuse-porous; between the age of 3 and 9 years, wood was semi-ring-porous; and between the age of 10 and 13 years, wood was ring-porous. The effect of clones on anatomical characteristics was significant except for the microfibril angle in latewood and ring width. The transition between juvenile and mature wood was between 7 and 8 years based on patterns of radial variation in fiber length (earlywood) and microfibril angle. From the pith to the bark, fiber length, double wall thickness, fiber wall: lumen ratio, vessel diameter in earlywood, proportion of vessel in earlywood, and axial parenchyma in latewood increased significantly, whereas ring width, earlywood vessels, and the proportion of fiber decreased significantly. In addition, other features, such as vessel length, microfibril angle, and ray proportion, did not differ significantly from the pith to the bark. Conclusions: Breeding program must consider both clone and cambial age to improve the economic profitability of wood production.

Tree physiological monitoring of the 2018 larch budmoth outbreak: preference for leaf recovery and carbon storage over stem wood formation in Larix decidua.

Insect defoliation impacts forest productivity worldwide, highlighting the relevance of plant-insect interactions. The larch budmoth (Zeiraphera griseana Hübner) is one of the most extensively studied defoliators, where numerous tree ring-based analyses on its host (Larix decidua Mill.) have aided in identifying outbreak dynamics over the past millennia. Yet, outbreaks have been widely absent after the early 1980s, and little is known about the in situ tree physiological responses and the allocation of carbon resources during and after defoliation. In summer 2018, we tracked an ongoing larch budmoth outbreak in a well-studied larch forest in the Swiss Alps. We performed biweekly monitoring on an affected and unaffected site using a unique combination of xylogenesis observations, measurements of non-structural carbohydrates, isotopic analysis of needle assimilates and ground-based and remote-sensed leaf trait observations. The budmoth induced a defoliation that lasted 40days and could be detected by satellite observations. Soluble sugars significantly decreased in needles and stem phloem of the defoliated trees, while starch levels remained stable in the stem and root xylem compared to the control. Carbon and oxygen isotope ratios in needle assimilates indicated that neither photosynthetic assimilation rates nor stomatal conductance was different between sites before, during and after the outbreak. Defoliated trees ceased cell wall thickening 17days earlier than unaffected trees, showing the earliest halt of ring formation recorded from 2007 untill 2013 and causing significant thinner cell walls, particularly in the latewood. No significant differences were found for cell enlargement rates and ring width. Our study revealed that an outbreak causes a downregulation of cell wall thickening first, while no starch is mobilized or leaf physiology is adjusted to compensate for the reduced carbon source due to defoliation. Our observations suggest that affected larch trees prioritize leaf recovery and carbon storage over wood biomass development.

DNA methylation and its effects on gene expression during primary to secondary growth in poplar stems

BackgroundAs an important epigenetic mark, 5-methylcytosine (5mC) methylation is involved in many DNA-dependent biological processes and plays a role during development and differentiation of multicellular organisms. However, there is still a lack of knowledge about the dynamic aspects and the roles of global 5mC methylation in wood formation in tree trunks. In this study, we not only scrutinized single-base resolution methylomes of primary stems (PS), transitional stems (TS), and secondary stems (SS) of Populus trichocarpa using a high-throughput bisulfite sequencing technique, but also analyzed the effects of 5mC methylation on the expression of genes involved in wood formation.ResultsThe overall average percentages of CG, CHG, and CHH methylation in poplar stems were ~ 53.6%, ~ 37.7%, and ~ 8.5%, respectively, and the differences of 5mC in genome-wide CG/CHG/CHH contexts among PS, TS, and SS were statistically significant (p < 0.05). The evident differences in CG, CHG, and CHH methylation contexts among 2 kb proximal promoters, gene bodies, and 2 kb downstream regions were observed among PS, TS, and SS. Further analysis revealed a perceptible global correlation between 5mC methylation levels of gene bodies and transcript levels but failed to reveal a correlation between 5mC methylation levels of proximal promoter regions and transcript levels. We identified 653 and 858 DMGs and 4978 and 4780 DEGs in PS vs TS and TS vs SS comparisons, respectively. Only 113 genes of 653 DMGs and 4978 DEGs, and 114 genes of 858 DMGs and 4780 DEG were common. Counterparts of some of these common genes in other species, including Arabidopsis thaliana, are known to be involved in secondary cell wall biosynthesis and hormone signaling. This indicates that methylation may directly modulate wood formation genes and indirectly attune hormone signaling genes, which in turn impact wood formation.ConclusionsDNA methylation only marginally affects pathway genes or regulators involved in wood formation, suggesting that further studies of wood formation should lean towards the indirect effects of methylation. The information and data we provide here will be instrumental for understanding the roles of methylation in wood formation in tree species.

Transcriptomic Profiling of Cryptomeria fortunei Hooibrenk Vascular Cambium Identifies Candidate Genes Involved in Phenylpropanoid Metabolism

Cryptomeria fortunei Hooibrenk (Chinese cedar) is a coniferous tree from southern China that has an important function in landscaping and timber production. Lignin is one of the key components of secondary cell walls, which have a crucial role in conducting water and providing mechanical support for the upward growth of plants. It is mainly biosynthesized via the phenylpropanoid metabolic pathway, of which the molecular mechanism remains so far unresolved in C. fortunei. In order to obtain further insight into this pathway, we performed transcriptome sequencing of the C. fortunei cambial zone at 5 successive growth stages. We generated 78,673 unigenes from transcriptome data, of which 45,214 (57.47%) were successfully annotated in the non-redundant protein database (NR). A total of 8975 unigenes were identified to be significantly differentially expressed between Sample_B and Sample_A after analyzing their expression profiles. Of the differentially expressed genes (DEGs), 6817 (75.96%) and 2158 (24.04%) were up- and down-regulated, respectively. 83 DEGs were involved in phenylpropanoid metabolism, 37 DEGs that encoded v-Myb avian myeloblastosis viral oncogene homolog (MYB) transcription factor (TF), and many candidates that encoded lignin synthesizing enzymes. These findings contribute to understanding the expression pattern of C. fortunei cambial zone transcriptome. Furthermore, our results provide additional insight towards understanding the molecular mechanisms of wood formation in C. fortunei.

Transcriptome analysis of salt-responsive and wood-associated NACs in Populus simonii \xd7\xa0Populus nigra

BackgroundNAC (NAM, ATAF1–2, and CUC2) family is one of the largest plant-specific transcription factor families known to play significant roles in plant development processes and stress responses.ResultsIn the study, a total of 112 NACs were identified to be differentially expressed in the comparisons of leaves and stems, leaves and roots, roots and stems of Populus simonii×P. nigra among 289 members by RNA-Seq. And 148, 144 and 134 NACs were detected to be salt-responsive in the roots, stems and leaves under 150 mM NaCl stress, respectively. Among them, a total of 53 salt-responsive NACs were shared across the three tissues. Under salt stress, 41/37 NACs were identified to be up/down-regulated in the leaves of Populus simonii × P.nigra among 170 non-redundant NACs by RT-qPCR, which was similar with RNA-Seq results. The expression pattern analysis of 6 NACs including four randomly up-regulated genes (NAC86, NAC105, NAC139 and NAC163) and two down-regulated genes (NAC15 and NAC149) indicated a few NACs showed specific temporal and spatial expression patterns in the three tissues of Populus simonii×P.nigra. Based on transcriptome screening and phylogenic analysis of differentially expressed NACs in different tissues under salt stress, 18 potential NACs associated with wood formation and 20 involved in stress responses were identified in Populus simonii×P.nigra.ConclusionsThe study further gains an understanding of the connection of tissue specificity and gene function in poplar, and lays the foundation of functional analysis of poplar NACs in stress responses.

Regulation of Lignin Biosynthesis by Post-translational Protein Modifications.

Post-translational modification of proteins exerts essential roles in many biological processes in plants. The function of these chemical modifications has been extensively characterized in many physiological processes, but how these modifications regulate lignin biosynthesis for wood formation remained largely unknown. Over the past decade, post-translational modification of several proteins has been associated with lignification. Phosphorylation, ubiquitination, glycosylation, and S-nitrosylation of transcription factors, monolignol enzymes, and peroxidases were shown to have primordial roles in the regulation of lignin biosynthesis. The main discoveries of post-translational modifications in lignin biosynthesis are discussed in this review.

Identification of putative lignin biosynthesis genes in Betula pendula

We identified 15 genes encoding enzymes for phenylpropanoid biosynthesis in the genome of birch by combining phylogenetic analysis and tissue-specific expression data. Lignin is one of the most abundant terrestrial biopolymers and is essential for plant structure and defense. An essential step in lignin formation is phenylpropanoid synthesis, we identified 120 gene models in 10 protein families encoding enzymes for phenylpropanoid biosynthesis in the genome of Betula pendula. The transcript abundance was determined for all 120 genes in xylem, root, leaf, and flower tissues using RNA-seq technology. We identified 15 genes that likely encode phenylpropanoid biosynthesis enzymes during wood formation. Ten of these genes are evolutionarily conserved compared to the lignin genes in Populus trichocarpa.

The Osmotin-Like Protein Gene PdOLP1 Is Involved in Secondary Cell Wall Biosynthesis during Wood Formation in Poplar.

Osmotin-like proteins (OLPs) mediate defenses against abiotic and biotic stresses and fungal pathogens in plants. However, no OLPs have been functionally elucidated in poplar. Here, we report an osmotin-like protein designated PdOLP1 from Populus deltoides (Marsh.). Expression analysis showed that PdOLP1 transcripts were mainly present in immature xylem and immature phloem during vascular tissue development in P. deltoides. We conducted phenotypic, anatomical, and molecular analyses of PdOLP1-overexpressing lines and the PdOLP1-downregulated hybrid poplar 84K (Populus alba × Populus glandulosa) (Hybrid poplar 84K PagOLP1, PagOLP2, PagOLP3 and PagOLP4 are highly homologous to PdOLP1, and are downregulated in PdOLP1-downregulated hybrid poplar 84K). The overexpression of PdOLP1 led to a reduction in the radial width and cell layer number in the xylem and phloem zones, in expression of genes involved in lignin biosynthesis, and in the fibers and vessels of xylem cell walls in the overexpressing lines. Additionally, the xylem vessels and fibers of PdOLP1-downregulated poplar exhibited increased secondary cell wall thickness. Elevated expression of secondary wall biosynthetic genes was accompanied by increases in lignin content, dry weight biomass, and carbon storage in PdOLP1-downregulated lines. A PdOLP1 coexpression network was constructed and showed that PdOLP1 was coexpressed with a large number of genes involved in secondary cell wall biosynthesis and wood development in poplar. Moreover, based on transcriptional activation assays, PtobZIP5 and PtobHLH7 activated the PdOLP1 promoter, whereas PtoBLH8 and PtoWRKY40 repressed it. A yeast one-hybrid (Y1H) assay confirmed interaction of PtoBLH8, PtoMYB3, and PtoWRKY40 with the PdOLP1 promoter in vivo. Together, our results suggest that PdOLP1 is a negative regulator of secondary wall biosynthesis and may be valuable for manipulating secondary cell wall deposition to improve carbon fixation efficiency in tree species.

Xylogenesis of Scots Pine in an Uneven-Aged Stand of the Minusinsk Depression (Southern Siberia)

Tree-ring formation studies are important for assessing the impact of environmental factors on tree growth at intra-seasonal resolution. This information is necessary for understanding plant acclimatization to current and expected climate changes. Little is still known about how tree age may affect the duration and rate of annual ring formation. In this study, we investigated tree-ring formation in Scots pine (Pinus sylvestris L.) trees of different ages (30- and 95-year-old trees) from the foreststeppe zone in Southern Siberia. The main objectives were 1) to estimate the timing of cambial activity by distinguishing the phases of division, enlargement, wall thickening, and maturation of tracheids and 2) to compare the anatomical structure of the tracheids forming the annual rings of the differently aged trees. Stem tissue was sampled weekly from April to September 2014. The results showed a 1-2 week difference in duration of the phases of xylem formation between the groups; in addition, the ring width of the young trees was slightly narrower. The size of the tracheids of the entirely formed ring (i.e. the results of the enlargement phase) did not differ between the groups whereas the dynamics of the cell-wall thickness showed significant differences. The data obtained in the present study can provide references to calibrate process-based models linking environment to wood formation. These data can be used to benchmark time-explicit simulated measurements of annual ring increment and cell anatomical structure against the corresponding parameters of mature trees growing under natural conditions