Tracheid Cell Walls Research Articles

Norway spruce (Picea abies) laccases: characterization of a laccase in a lignin-forming tissue culture.

Secondarily thickened cell walls of water-conducting vessels and tracheids and support-giving sclerenchyma cells contain lignin that makes the cell walls water impermeable and strong. To what extent laccases and peroxidases contribute to lignin biosynthesis in muro is under active evaluation. We performed an in silico study of Norway spruce (Picea abies (L.) Karst.) laccases utilizing available genomic data. As many as 292 laccase encoding sequences (genes, gene fragments, and pseudogenes) were detected in the spruce genome. Out of the 112 genes annotated as laccases, 79 are expressed at some level. We isolated five full-length laccase cDNAs from developing xylem and an extracellular lignin-forming cell culture of spruce. In addition, we purified and biochemically characterized one culture medium laccase from the lignin-forming cell culture. This laccase has an acidic pH optimum (pH 3.8-4.2) for coniferyl alcohol oxidation. It has a high affinity to coniferyl alcohol with an apparent Km value of 3.5 μM; however, the laccase has a lower catalytic efficiency (V(max)/K(m)) for coniferyl alcohol oxidation compared with some purified culture medium peroxidases. The properties are discussed in the context of the information already known about laccases/coniferyl alcohol oxidases of coniferous plants.

MICROSTRUCTURE DESCRIPTION AND MICROMECHANICAL PROPERTIES OF SPRUCE WOOD

The knowledge about the microstructure and morphology of individual phases also allows an artificial modification of the material, or its processing, to produce engineered products with required properties. The purpose of our work was to investigate the distribution of elastic stiffness within the tissues of individual cells using the finest equipment and to observe the morphology of individual phases. A quasi-static nanoindentation was carried out on the cell walls of earlywood and latewood tracheids of spruce wood. The dynamic modulus mapping, also known as nanoDMA, was utilized to obtain the map of elastic moduli over the entire tracheid cross-section. In particular, it was found that the stiffness cells walls is approximately 10.5 GPa and 12.5 GPa in the case of earlywood and latewood tracheids, respectively. The difference between earlywood and latewood elastic stiffness is attributed to a different chemical composition and orientation of fibrils. The acquired data are indispensible for micromechanical modeling and design of engineered products with superior mechanical properties.

Hydrogel-based boron salt formulations for wood preservation

Mixing boron with two thermoreversible hydrogels, obtained from an amphiphilic tripeptide and a carbamate derivative of glycerol, respectively, allows the formation of a supramolecular network that incorporates boron and significant amounts of water upon gelification of the solution when the temperature decreases. Hydrogels obtained from these gelators were impregnated in pine sapwood blocks using vacuum pressure treatment and subjected to leaching. Results indicated that incorporation of boron salts in the hydrogel network effectively protected the wood from degradation caused by the brown rot fungus Poria placenta and from attack by termites also even after leaching. It is believed that these hydrogels fill the tracheid cell walls and lumens, limiting the leachability of boron salts when the wood is humidified.

Wood defects during industrial-scale production of thermally modified Norway spruce and Scots pine

This research investigates wood defects, particularly the formation of surface cracks, during the production of thermally modified wood and its exposure to cyclic moisture changes. Boards of Norway spruce and Scots pine originating from different steps within the production of ThermoWood® were collected and wood defects were investigated at macroscopic and microscopic scale. Subsequently, the wood was exposed to capillary wetting cycles to record its sensitivity towards cracking. After the modification process, typical anatomical defects of conventional kiln-drying became more frequent and severe, with the magnitude being to some extent depending on the presence of defects in the raw material. At microscopic scale, damages to ray parenchyma and epithelial cells as well as longitudinal cracks within the cell walls of earlywood tracheids were evident in thermally modified wood. Despite a lower water uptake and higher dimensional stability, thermally modified wood was more sensitive to surface cracking during wetting cycles than unmodified wood, i.e. at the outside face of outer boards (near bark). For limiting surface cracking of thermally modified wood during service life, the use of high-quality raw material, the exposure of the inside face of the boards (near pith) and the application of a surface coating are considered beneficial.

Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay.

Compression wood is a type of reaction wood formed on the underside of softwood stems when they are tilted from the vertical and on the underside of branches. Its quantification is still a matter of some scientific debate. We developed a new technique that has the potential to do this based on the higher proportions of (1→4)-β-d-galactans that occur in tracheid cell walls of compression wood. Wood was milled, partially delignified, and the non-cellulosic polysaccharides, including the (1→4)-β-d-galactans, extracted with 6 M sodium hydroxide. After neutralizing, the solution was serially diluted, and the (1→4)-β-d-galactans determined by an immuno-dot assay using the monoclonal antibody LM5, which specifically recognizes this polysaccharide. Spots were quantified using a dilution series of a commercially available (1→4)-β-d-galactan from lupin seeds. Using this method, compression and opposite woods from radiata pine (Pinus radiata) were easily distinguished based on the amounts of (1→4)-β-d-galactans extracted. The non-cellulosic polysaccharides in the milled wood samples were also hydrolysed using 2 M trifluoroacetic acid followed by the separation and quantification of the released neutral monosaccharides by high performance anion exchange chromatography. This confirmed that the compression woods contained higher proportions of galactose-containing polysaccharides than the opposite woods.

Evidence for Systemic Infection by Puccinia horiana, Causal Agent of Chrysanthemum White Rust, in Chrysanthemum.

Puccinia horiana, causal agent of the disease commonly known as chrysanthemum white rust (CWR), is a quarantine-significant fungal pathogen of chrysanthemum in the United States and indigenous to Asia. The pathogen was believed to have been eradicated in the United States but recently reappeared on several occasions in northeastern United States. The objective of the study presented here was to determine whether P. horiana could systemically infect chrysanthemum plants, thus providing a means of survival through winters. Scanning and transmission electron microscopy revealed the development of P. horiana on the surface and within leaves, stems, or crowns of inoculated chrysanthemum plants artificially exposed to northeastern U.S. winter temperatures. P. horiana penetrated leaves directly through the cuticle and then colonized the mesophyll tissue both inter- and intracellularly. An electron-dense material formed at the interface between fungal and host mesophyll cells, suggesting that the pathogen adhered to the plant cells. P. horiana appeared to penetrate mesophyll cell walls by enzymatic digestion, as indicated by the absence of deformation lines in host cell walls at penetration sites. The fungus was common in vascular tissue within the infected crown, often nearly replacing the entire contents of tracheid cell walls. P. horiana frequently passed from one tracheid cell to an adjacent tracheid cell by penetration either through pit pairs or nonpitted areas of the cell walls. Individual, presumed, fungal cells in mature tracheid cells of the crown and stems arising from infected crowns suggested that the pathogen might have been moving at least partially by means of the transpiration stream. The demonstration that chrysanthemum plants can be systemically infected by P. horiana suggests that additional disease control measures are required to effectively control CWR.

Investigation of Low-molecular Weight Phenol Formaldehyde Distribution in Tracheid Cell Walls of Chinese Fir Wood

Treatment with water-soluble low-molecular weight phenol-formaldehyde resin is an effective method to improve wood properties. In this paper, plantation wood of Chinese fir was modified with low-molecular weight phenol-formaldehyde resin. The absorbance by tracheid cell walls of phenol-formaldehyde resin in treated and untreated reference samples were measured with an ultraviolet micro-spectrophotometer. The UV absorbance values of earlywood tracheids and middle lamella in treated wood were significantly increased, with an average increase of 49% and 23%, respectively. Moreover, after treatment with low-molecular weight phenol-formaldehyde resin, the UV absorbance of the earlywood tracheid cell walls of Chinese fir increased to more than 47%, regardless of whether or not the cell lumens were filled with resin. After treatment with low-molecular weight phenol-formaldehyde resin, the UV absorbance of earlywood tracheid cell walls at different locations did not vary greatly. This study provides direct support for the improvement of the physical and mechanical properties of resin-modified Chinese fir in terms of penetration of the resin into the cell walls.

Seasonal and clonal variation in cellulose microfibril orientation during cell wall formation of tracheids in Cryptomeria japonica

To investigate the biological mechanism by which trees control the changes in microfibril (MF) orientation among secondary cell wall layers of conifer tracheids, we studied seasonal variation in the orientation of newly deposited MFs during tracheid cell wall development in Japanese cedar (Cryptomeria japonica D. Don) trees growing in Central Japan (36°36'N, 140°39'E). Sample blocks were repeatedly collected from four 16-year-old clones of different origins during the growing season of 2010 to investigate the hypotheses that changes in cellulose MF orientation between wall layers exhibited seasonal and clonal differences. The progressive change in the orientation of newly deposited MFs on the primary and secondary cell wall layers of tracheids was detected by field-emission-scanning electron microscopy. Tracheid production and differentiation was studied by light microscopy. We observed a decreasing trend in the orientation of deposited MFs from earlywood to latewood in the S2 and S1 layers, where MFs appeared in a Z-helix. In contrast, no seasonal pattern in the orientation of the MFs in the S-helix was observed. Minor clonal variation was observed in the phenology of tracheid production and differentiation. We concluded that a seasonal decreasing trend in the orientation of the MFs in the Z-helix in S1 and S2 was present, whereas the MFs in other layers exhibited minor random variations. Thus, the orientation of the MFs in S2 was affected by seasonal factors, whereas the MFs in other layers were more intrinsically controlled. The within-ring variations in the MF orientation and thus the resulting average MF angle might also be related to genotypic differences in the tracheid production and differentiation rate. However, our results do not exclude other intrinsic and environmental regulations in the change in MF orientation, which remains a topic for future studies.

Reaction Wood Anatomy in a Vessel-Less Angiosperm Sarcandra Glabra

Anatomy and lignin distribution in artificially inclined stems of Sarcandra glabra were investigated to clarify the characteristics of reaction wood (RW) in a vessel-less angiosperm species. Of the five coppiced stems studied from a single tree, two stems were fixed straight and classified as normal wood (NW) and the remaining three stems were inclined at 50 degrees from the vertical to induce the formation of the RW. Compared with NW, the lower side of the inclined samples had a relatively high compressive surface-released strain and an increase in the microfibril angle of the S2 layer of tracheids. However, no significant change was observed in the length or cell wall thickness of the tracheids. The results of Wiesner and Mäule colour reactions indicated that the amount of guaiacyl lignin in the cell walls of tracheids was increased in RW. It appears that RW in Sarcandra is formed on the lower side of inclined stems, and its anatomical characteristics and chemical composition are similar to those of the compression wood (CW) found in gymnosperm species (the so-called “CW-like RW” type).

Modeling Microfibril Angle and Tree Age in <i>Acacia</i> <i>mangium</i> Wood Using X-Ray Technique

The term microfibril angle (MFA) in wood refers to the angle between the spiralling cellulose fibrils and the long axis of the tracheid cell wall. Diffraction patterns arising from crystal planes of various sample forms of wood trees had attracted scientific research in determining the crystallographic measurements. As such the tropical hard wood in Sabah, Acacia mangium was chosen for experimental data. Age-contributing factors were measured; the angle of reflection (θ), relative intensity, full width at half maximum (FWHM), the nearest between two neighbouring atoms in the crystalline structure (d-spacing) and the peak height, had been taken into account at different ages, pith and bark of tree. Regressions were done in comparing the microfibril angle, MFA at different ages using the least-square method and cubic-spline interpolation. The latter was able to interpolate a polynomial up to the third order. The range of the optimum angle was found to have benefited foresters in deciding the time for tree cropping and harvesting.

Comparison of Anatomy and Composition Distribution between Normal and Compression Wood of Pinus Bungeana Zucc. Revealed by Microscopic Imaging Techniques

The anatomy and topochemistry in normal and compression wood tracheid cell wall of Pinus bungeana Zucc. were investigated by fluorescence microscopy and confocal Raman microscopy. Using fluorescence microscopy, the severity of compression wood was classed as a mild type for the reason that it did not contain all compression wood features. Chemical imaging by confocal Raman microscopy was used for analyzing the distribution of lignin and cellulose, as well as the functional groups of lignin in tracheid cell walls. By comparison with normal wood, highly lignified outer S2 layer [S2(L)], thicker S1 layer, and obviously reduced lignification in the middle lamella were characteristic of compression wood. In addition, smaller microfibril angle was observed in the S2(L) region. The distribution of coniferyl alcohol and coniferyl aldehyde in normal and compression wood was enriched in S1 and S2 layers but lack in cell corner and/or S2L regions, which showed an opposite pattern to lignin distribution. Confocal Raman microscopy with high spatial resolution contributes to a further understanding of the differences between normal and compression wood in polymers distribution and molecules orientation in situ.

Effect of Modification with Phenol Formaldehyde Resin on the Mechanical Properties of Wood from Chinese Fir

Samples of Chinese fir were treated with either low-molecular-weight or commercial phenol-formaldehyde (PF) resins. The macro- and micromechanical properties of the treated and untreated samples were determined. The average longitudinal tensile modulus of elasticity (MOE) was 30.88% larger for the samples treated with the low-molecular-weight PF resin than it was for the untreated samples. The average MOE of the samples treated with the commercial PF resin was 29.84% less than that of the untreated samples. The micromechanical properties of the samples were investigated through nanoindentation studies. For the samples modified with low-molecular-weight PF resin, the values of average MOE and hardness were 32.94 and 32.93%, respectively, greater than those of the untreated samples. In contrast, the average MOE and hardness values were 11.99 and 18.14%, respectively, greater for the samples modified with commercial PF resin compared to the untreated samples. It could be inferred that the low-molecular-weight PF resin was able to diffuse into the nanopores in the S2 layer of the tracheid cell wall of the Chinese fir, thereby improving its macromechanical properties. Modification with low-molecular-weight PF resin was an effective way to enhance the longitudinal macromechanical properties of wood from the Chinese fir.

Correlative light and scanning electron microscopy of the same sections gives new insights into the effects of pectin lyase on bordered pit membranes in Pinus radiata wood

Bordered pits are structures in the cell walls of softwood tracheids which permit the movement of water between adjacent cells. These structures contain a central pit membrane composed of an outer porous ring (margo) and an inner dense and pectin-rich disc (torus). The membrane is overarched on each side by pit borders. Pits may be aspirated, a condition where the torus seals against the pit border, effectively blocking the pathway between cells. In living trees this maintains overall continuity of water conduction in xylem by sealing off tracheids containing air. Drying of timber results in further pit aspiration, which reduces wood permeability to liquid treatment agents such as antifungal chemicals. One possible way to increase permeability is by treating wood with pectin lyase to modify or remove the torus. The effectiveness of this treatment was initially evaluated using light microscopy (LM) of toluidine blue stained wood. Pectic material is coloured pink-magenta with this stain, and loss of this colour after treatment has been interpreted as indicating destruction of the torus. However, correlative light (LM) and scanning electron (SEM) microscopic observations of identical areas of toluidine blue stained sections revealed that many unstained pits had intact but modified tori when viewed with SEM. These observations indicate that LM alone is not sufficient to evaluate the effects of pectin lyase on pit membranes in wood. Combining LM and SEM gives more complete information

Modeling Microfibril Angle and Tree Age in Acacia Mangium Wood Using X-Ray Diffraction Technique

 Abstract—The term microfibril angle (MFA) in wood refers to the angle between the spiralling cellulose fibrils and the long axis of the tracheid cell wall. Diffraction patterns arising from crystal planes of various sample forms of wood trees had attracted scientific research in determining the crystallographic measurements. Acacia mangium classified as a hardwood was chosen for experimental data. Age- contributing factors were measured; the angle of reflection (θ), relative intensity, full width at half maximum (FWHM), the nearest between two neighbouring atoms in the crystalline structure (d-spacing) and the peak height, had been taken into account at different ages, pith and bark of tree. Regressions were done in comparing the microfibril angle, MFA at different ages using the least-square method and cubic-spline interpolation. The latter was able to interpolate a polynomial up to the third order. The range of the optimum angle was found to have benefited foresters in deciding the time for tree cropping and harvesting.

Ultrastructure of the innermost surface of differentiating normal and compression wood tracheids as revealed by field emission scanning electron microscopy

The ultrastructure of the innermost surface of Cryptomeria japonica differentiating normal wood (NW) and compression wood (CW) was comparatively investigated by field emission electron microscopy (FE-SEM) combined with enzymatic degradation of hemicelluloses. Cellulose microfibril (CMF) bundles were readily observed in NW tracheids in the early stage of secondary cell wall formation, but not in CW tracheids because of the heavy accumulation of amorphous materials composed mainly of galactans and lignin. This result suggests that the ultrastructural deposition of cell wall components in the tracheid cell wall differ between NW and CW from the early stage of secondary cell wall formation. Delignified NW and CW tracheids showed similar structural changes during differentiating stages after xylanase or β-mannanase treatment, whereas they exhibited clear differences in ultrastructure in mature stages. Although thin CMF bundles were exposed in both delignified mature NW and CW tracheids by xylanase treatment, ultrastructural changes following β-mannanase treatment were only observed in CW tracheids. CW tracheids also showed different degradation patterns between xylanase and β-mannanase. CMF bundles showed a smooth surface in delignified mature CW tracheids treated with xylanase, whereas they had an uneven surface in delignified mature CW tracheids treated with β-mannanase, indicating that the uneven surface of CMF bundles was related to xylans. The present results suggest that ultrastructural deposition and organization of lignin and hemicelluloses in CW tracheids may differ from those of NW tracheids.

Localization of Cell Wall Polysaccharides in Normal and Compression Wood of Radiata Pine: Relationships with Lignification and Microfibril Orientation

The distribution of noncellulosic polysaccharides in cell walls of tracheids and xylem parenchyma cells in normal and compression wood of Pinus radiata, was examined to determine the relationships with lignification and cellulose microfibril orientation. Using fluorescence microscopy combined with immunocytochemistry, monoclonal antibodies were used to detect xyloglucan (LM15), β(1,4)-galactan (LM5), heteroxylan (LM10 and LM11), and galactoglucomannan (LM21 and LM22). Lignin and crystalline cellulose were localized on the same sections used for immunocytochemistry by autofluorescence and polarized light microscopy, respectively. Changes in the distribution of noncellulosic polysaccharides between normal and compression wood were associated with changes in lignin distribution. Increased lignification of compression wood secondary walls was associated with novel deposition of β(1,4)-galactan and with reduced amounts of xylan and mannan in the outer S2 (S2L) region of tracheids. Xylan and mannan were detected in all lignified xylem cell types (tracheids, ray tracheids, and thick-walled ray parenchyma) but were not detected in unlignified cell types (thin-walled ray parenchyma and resin canal parenchyma). Mannan was absent from the highly lignified compound middle lamella, but xylan occurred throughout the cell walls of tracheids. Using colocalization measurements, we confirmed that polysaccharides containing galactose, mannose, and xylose have consistent correlations with lignification. Low or unsubstituted xylans were localized in cell wall layers characterized by transverse cellulose microfibril orientation in both normal and compression wood tracheids. Our results support the theory that the assembly of wood cell walls, including lignification and microfibril orientation, may be mediated by changes in the amount and distribution of noncellulosic polysaccharides.

Effect of Physisporinus vitreus on wood properties of Norway spruce. Part 2: Aspects of microtensile strength and chemical changes

AbstractThe biotechnological application of the white rot fungusPhysisporinus vitreusnamed “bioincising” is currently being investigated for permeability improvement of Norway spruce (Picea abies(L.) Karst.) wood. During short-term (<9 weeks) incubation, fungal activity induces degradation of pit membranes and a simultaneous alteration of the tracheid cell wall structure. In Part 1 of this article series, the occurrence of selective delignification and simultaneous degradation was shown by UV-microspectrophotometry (UMSP). Moreover, significant reduction of Brinell hardness was recorded after 7 and 9 weeks incubation. For a better understanding of the chemical alterations in the wood constituents and the corresponding changes of mechanical properties due to fungal activity, we applied microtensile tests on thin strips that were prepared from the surface of incubated Norway spruce wood. Indications for the occurrence of selective delignification and simultaneous degradation were evident. Determination of lignin content and carbohydrate analysis by borate anion exchange chromatography confirmed the results. The present study verifies the findings from Part 1 of this article series and from previously conducted microscopic investigations. Now, the degradation characteristics ofP. vitreusare established and the bioincising process can be further optimized with higher reliability.

Effect of Physisporinus vitreus on wood properties of Norway spruce. Part 1: Aspects of delignification and surface hardness

Abstract The white rot fungus Physisporinus vitreus is currently tested for several biotechnological applications such as permeability improvement of refractory wood species or the optimization of the acoustic properties of wood for violins. The enzymatic activity of P. vitreus results in the degradation of pit membranes and simultaneous alterations of the tracheid cell wall structure in wood of Norway spruce [Picea abies (L.) Karst]. By this means, selective delignification and simultaneous degradation occur in the latewood tracheids at short incubation times. To study the delignification of individual cell wall layers in latewood tracheids, cellular UV-microspectrophotometry was applied to wood of Norway spruce that had been incubated for between 3 and 9 weeks. By means of this technique, the progressing delignification was demonstrated in the latewood tracheid secondary walls. Moreover, local delignification in close proximity to hyphal tunneling, cavities, and notches was evident. Additionally, the mechanical changes were measured (a) at the macroscopic level by Brinell hardness test and (b) at the cellular level by nanoindentation. Brinell hardness was significantly reduced with increasing incubation time which was attributed to the partial delignification. Unlike Brinell tests, results from nanoindentation tests did not show a clear effect of fungal activity because of the material heterogeneity and the high spatial resolution of this technique. The present study provides methodological approaches for the investigation of wood-fungus interactions and contributes to a better understanding of the characteristics of wood decay at the subcellular level caused by the white rot fungus P. vitreus. Moreover, it establishes the basis for a subsequent chemical analysis, for which the results will be the topic of a second paper in this series.

Microfibril organization modes in plant cell walls of variable curvature: a model system for two dimensional anisotropic soft matter

We present a mechanical model of the plant cell wall viewed as anisotropic two dimensional soft matter, where a dilute dispersion of cellulose fibrils of variable orientations is uniformly distributed on a curved deformable viscoelastic matrix membrane. The plant cell wall model integrates the elastic energy of the curved membrane, the nematic Landau–de Gennes fiber orientation energy, and competing curvophilic and curvophobic interactions mediated by the membrane geometry and the fibrils' orientation. The selected membrane geometry is a straight cylinder of variable cross-section, whose shape varies from a circle to a sharp super-ellipse as in many cell specimens, and whose size increases as in plant cell wall growth. Model predictions indicate that due to curvature–orientation couplings, the fiber orientation displays three modes: (i) line (along the cylindrical axis), (ii) helical (oblique to the axis), and (iii) ring (normal to the axis), where the former arises under large curvature and the latter under small one. In addition to aligning the fibers in the line mode, high curvature also promotes the order of the fibrils. The predicted fiber structure is validated with fiber structures in the cell wall of tracheids. The structure–property relations for super-elliptical membranes with gradient curvature are established and its role in cell growth shape is predicted. The principal mechanisms are the role of fiber orientation and order on bending stiffness: (a) orientation along the axis has no stiffening effect, (b) orientation along the azimuthal direction produces maximal stiffening, and (c) fiber randomization softens the membrane. The largest fiber stiffening effect is found when the membrane length scale (radius of curvature) and fiber length scale (curvophobic/curvophilic energy ratio) are equal. It is found that super-elliptical shape invariant growth and expansion is preferred for cells with sharp/soft corners and straight/stiff sides. Otherwise growth promotes shape changes. Lastly, master plots that categorize fiber structures by the number of multimodal regimes (line/helix, line/helix/ring) that arise in super-ellipses are used to relate shape and size to domain structures that affect toughness and modulus.

Immunolocalization and structural variations of xylan in differentiating earlywood tracheid cell walls of Cryptomeria japonica

We investigated the spatial and temporal distribution of xylans in the cell walls of differentiating earlywood tracheids of Cryptomeria japonica using two different types of monoclonal antibodies (LM10 and LM11) combined with immunomicroscopy. Xylans were first deposited in the corner of the S(1) layer in the early stages of S(1) formation in tracheids. Cell corner middle lamella also showed strong xylan labeling from the early stage of cell wall formation. During secondary cell wall formation, the innermost layer and the boundary between the S(1) and S(2) layers (S(1)/S(2) region) showed weaker labeling than other parts of the cell wall. However, mature tracheids had an almost uniform distribution of xylans throughout the entire cell wall. Xylan localization labeled with LM10 antibody was stronger in the outer S(2) layer than in the inner layer, whereas xylans labeled with LM11 antibody were almost uniformly distributed in the S(2) layer. In addition, the LM10 antibody showed almost no xylan labeling in the S(1)/S(2) region, whereas the LM11 antibody revealed strong xylan labeling in the S(1)/S(2) region. These findings suggest that structurally different types of xylans may be deposited in the tracheid cell wall depending on the developmental stage of, or location in, the cell wall. Our study also indicates that deposition of xylans in the early stages of tracheid cell wall formation may be spatially consistent with the early stage of lignin deposition in the tracheid cell wall.