Xylem cells are surrounded by primary and secondary cell walls. Formation of primary walls is regulated by the cell wall integrity surveillance system, but it is unclear if the deposition of secondary walls is similarly regulated. To study this question, we introduced to aspen three different enzymes cleaving cell wall-localized xylan and we suppressed xylan synthase components either ubiquitously or specifically during secondary wall formation using the Populus trichocarpa GT43B promoter. When xylan was ubiquitously altered, 95% of lines showed reduced growth, whereas when it was altered during secondary wall deposition, 30% of lines grew better, with the rest having no growth impairment, suggesting opposite effects of primary and secondary wall disturbances. To detect the mechanism of growth stimulation by disturbed deposition of the secondary wall, we analyzed changes in wood quality traits, chemistry, transcriptomics, metabolomics and hormonomics in transgenic lines. We found increased tension wood production, reduced S- and H-lignin, and changes in several metabolites in common in these lines. Remorin REM1.3 and NRL2 (NPH3 family) transcripts increased, and changes in jasmonates, abscisic acid, and salicylic acid occurred in secondary wall-forming xylem, suggesting their involvement in secondary wall integrity surveyance and signaling. The data indicate that a unique program mediates responses to secondary wall impairment that induces growth.

Understanding the plant cell wall architecture is essential for elucidating its biological function and mechanical properties. This study employs a synthetic approach using spherical core-shell capsules with shells composed of deuterated bacterial cellulose (d-BC) and pectin. The shell structure was created via a bottom-up layer-by-layer assembly onto CaCO3 templates, followed by characterization through microscopy and scattering techniques. Small-angle X-ray scattering (SAXS) and confocal laser scanning microscopy revealed increased pore sizes in hydrated d-BC/pectin shells compared to those of hydrated wood-derived cellulose nanofiber (CNF)-based shells from a previous study. Using small-angle neutron scattering (SANS) with contrast variation, structural changes of individual wall components under varying salinities (0 or 10 mM NaCl) were analyzed. The presence of NaCl selectively influenced the pectin phase, while the d-BC network retained structural stability, highlighting its robustness as a wall component. This platform provides a useful tool for testing hypotheses and advancing our understanding of cell wall porosity and composition-dependent permeability.

Side streams from wheat processing, such as the bran and gluten fractions, show great potential as a feedstock for the production of novel food ingredients and materials. In this study, we prepared hybrid polysaccharide-protein hydrogels via enzymatic crosslinking of wheat bran arabinoxylan and gluten fractions. Arabinoxylan was first isolated from wheat bran via subcritical water extraction, which preserved the covalently bound ferulic acid moieties to the arabinoxylan core amenable for laccase crosslinking. Gluten was fractionated into its main protein components (glutenin and gliadin) via treatment with aqueous ethanol. Hydrogels with different contents of arabinoxylan and gluten were prepared, demonstrating the integration of the protein fractions within the polysaccharide gel network. Increased addition of gluten led to gradually softer hydrogels, suggesting that the gluten fractions were not involved in the covalent crosslinking with the ferulic acid moieties to any noticeable level. Freeze-drying and regeneration of the hydrogels led to a 3-fold-10-fold increase in the storage and loss moduli, depending on the sample. Analysis of the structure of the hydrogels revealed that the addition of gluten upon enzymatic crosslinking impacted the physical interactions and crystallinity of the arabinoxylan populations, resulting in phase separation of the protein and polysaccharide components. This study demonstrates that tunable hydrogels can be prepared from cereal side streams, with potential as functional plant-based food hydrocolloids with improved nutritional properties, combining dietary fibre and protein components.

Alterations in the structure and chemistry of cell wall polymers during wood decay by white-rot fungi could be one of the best experimental systems to studying the association between different cell wall polymers and the biology of plant-microbe interactions. We investigated the spatial and temporal changes in the distribution patterns of matrix polysaccharides and lignin in the fibre cell walls of D. sissoo wood subjected to preferential delignification and simultaneous decay by two species of white rot fungi. Transmission electron microscopy analysis of fibre walls affected with L. betulina showed removal of lignin from the S1 layer of the secondary walls (SW), resulting in cell separation. Subsequently, preferential removal of lignin from the S2 and S3 layers was observed. The structural changes in the SW of fibres inoculated with D. flavida directly correlated with the simultaneous degradation of all wall polymers. Immunogold labelling-TEM analysis revealed degradation of xyloglucan in the compound middle lamellae (CML) region, undergoing preferential delignification. Weak labelling for less substituted heteroxylans was evident in S2 and S3 layers of preferentially delignified fibre walls. Highly substituted heteroxylans showed a higher distribution in the outer layers of SW even at late stages of degradation. The degradation pattern of cell wall polymers suggests a close association between lignin-heteroxylans in the SW as they were removed simultaneously during preferential delignification in the xylem fibres.

Cuticle - a hydrophobic barrier of cutin and waxes covering the outer cell wall surface of plants - enables survival in terrestrial habitats. However, it is not understood how the hydrophobic cuticle precursors travel through the homogalacturonan-rich hydrophilic cell wall. To elucidate the role of homogalacturonan in cuticle development, we disrupted its integrity by overexpressing a pectate lyase, PtxtPL1-27, in aspen. PtxtPL1-27 had pleiotropic effects on shoot development, including the reduction of cuticle thickness and changes in cutin and wax composition, but the expression of cutin biosynthetic genes was little affected. Despite a reduction in homogalacturonan content in the leaves, labeling with the homogalacturonan-specific antibody JIM5 in the outer epidermal cell wall layer increased and displayed an altered pattern. Moreover, the ultrastructure of cell walls was changed concomitant with lipid accumulation. We propose that the disruption of homogalacturonan integrity affected the cutinsome-dependent transport and polymerization of cutin monomers in the cell wall.

Spatial information on wood structure and chemistry is crucial for understanding wood functionality. We present a high-throughput and high-resolution near-infrared (NIR) method for combined imaging of the physical and chemical properties of stem sections from Populus trees. Pyrolysis-GC/MS data was used for sensitive and spatially resolved calibration of wood chemistry while SilviScan'" analyses provided reference data for wood physical properties with 25 mu m resolution for wood density and 0.2-2.0 mm for microfibril angle (MFA). NIR prediction models were trained and calibrated on material from both fieldand greenhouse-grown trees. Thus, the method was developed for NIR imaging of stem samples as small as 4 mm in diameter with an image resolution of 0.03 mm for small-diameter samples and 0.5 mm for samples with multiple annual rings. The NIR model performance, tested against data not used in the training set, reached the coefficient of determination (R2pred) values for wood density and MFA of 0.60 and 0.72, respectively. The NIR models for wood chemistry showed R2pred values of 0.78 and 0.77 for carbohydrates and lignin, respectively. Models for the G-, S- and H-type lignin had R2pred values between 0.58 and 0.86. In addition, we developed a prediction model for the determination of tension wood distribution. According to this model, tension wood was frequently observed in young greenhouse samples, which might explain the higher variation found in the chemical and physical properties of wood in greenhouse-grown compared to field-grown trees. The study also demonstrated that NIR-model estimations in image format can capture spatial variations that are not detectable in bulk analyses of wood properties. Examples of the method applied to greenhouse-grown trees highlight the efforts to develop NIR models with good prediction accuracies based on high-resolution data.

Wood is the most abundant renewable natural resource composed of different polysaccharides and lignin, but its utilisation is hampered by intermolecular linkages between these components forming lignin-carbohydrate complexes (LCCs) causing recalcitrance. The links between glucuronoxylan and the γ-C of lignin (γ-ester linkages) are thought to contribute to one-third of LCCs, but direct evidence for their natural occurrence and their role in recalcitrance has been scarce so far. To address these issues, Phanerochaete carnosa glucuronoyl esterase (PcGCE), hydrolysing γ-ester linkages, was expressed in cell walls of developing wood in hybrid aspen (Populus tremula L. × tremuloides Michx.). The enzyme reduced HSQC 2D NMR signals corresponding to the γ-esters and xylan in dioxane-extracted LCCs without altering glucuronoxylan content or structure. This increased acid solubility of lignin and decreased lignin content. Reduced wood recalcitrance was shown by increased sugar yields and glucose production rates (by approx. 20%) in saccharification without pretreatment and increased xylan extractability by subcritical water (by approx. 70%). Moreover, trees expressing PcGCE exhibited greater primary and secondary growth. Transcriptomics and metabolomics analyses in developing wood suggested that growth could have been induced by a higher transcription of SMR2 and RPOTmp, which was likely triggered by the secondary cell wall integrity signalling. The results provide evidence for the natural existence of LCC γ-esters and their significant contribution to lignocellulose recalcitrance. Furthermore, they show that reducing γ-ester linkages could increase plant productivity.

Xylem cells are surrounded by primary and secondary cell walls. Formation of primary walls is regulated by the cell wall integrity surveyance system, but it is unclear if the deposition of secondary walls is similarly regulated. To study this question, we introduced to aspen three different enzymes cleaving cell wall-localized xylan and we suppressed xylan synthase components either ubiquitously or specifically during secondary wall formation. When xylan was ubiquitously altered, 95% of lines showed reduced growth, whereas when it was altered during secondary wall deposition, 30% of lines grew better with the rest having no growth impairment, suggesting opposite effects of primary and secondary wall disturbances. To detect mechanism of growth stimulation by disturbed deposition of secondary wall, we analyzed changes in wood quality traits, chemistry, transcriptomics, metabolomics and hormonomics in transgenic lines. We found increased tension wood production, reduced S- and H-lignin, and changes in several metabolites in common in these lines. Remorin REM1.3 and NRL2 ( NPH3 family) transcripts increased and changes in jasmonates, ABA and SA occurred in secondary wall-forming xylem suggesting their involvement in secondary wall integrity surveyance and signaling. The data indicate that a unique program mediates responses to secondary wall impairment that induces growth. Highlight Disturbance of the deposition of secondary walls that are responsible for the mechanical strength of plant bodies can increase growth with discrete changes in gene expression, hormone levels and metabolism. ### Competing Interest Statement The authors have declared no competing interest.

Living xylem fibres play important physiological and mechanical functions in trees. Despite a significant role of secondary cell wall chemical composition in performing these functions, little information is available on the developmental changes in the cell wall of living fibres. In the present study, the distribution pattern of pectins, hemicelluloses, and lignin in the cell walls during the differentiation and maturation of living fibres in the secondary xylem of Leucaena leucocephala was examined by light and electron microscopy. The expansion of primary walls during the early stage of fibre development was characterised by a change in the organisation of pectic polysaccharides in the middle lamellae region. The intercellular regions became filled with pectic polysaccharides following initiation of secondary wall deposition. Subsequently, lignification started at cell corners with the deposition of guaiacyl units that co-polymerise with syringyl moieties in the final stages of fibre development. The transmission electron microscopic analysis confirmed the disorganisation of pectic polysaccharides in the middle lamellae region during cell expansion and their inhomogeneous distribution in the cell corners following secondary wall deposition. Immunofluorescence microscopy revealed that β(1–4)-galactans are mainly incorporated in the middle lamellae region that undergoes disorganisation and reorganisation during and after cell expansion. Immunogold labelling experiments using JIM5, JIM7 and CCRCM1 antibodies revealed dynamic changes in the distribution pattern of homogalacturonan with different degrees of methylation and fucosylated xyloglucans during cell wall loosening and secondary wall maturation stages of xylem fibres. In mature fibres, LM10 labelling indicated that the less substituted xylans are distributed throughout the secondary wall, while labelling of highly substituted xylans with LM11 appeared more intense at the corner regions of the secondary wall compared to other regions. The KMnO4 staining revealed the relatively higher lignin distribution in xylem fibres in compound middle lamellae and S3 wall layers. The transition zone between S1 and S2 layers showed relatively high lignin distribution compared to the rest of the S2 wall layer. The ultrastructural studies demonstrated that the inhomogeneous distribution of lignin corresponds with that of pectins at the cell corners of fibres. The cell wall delignification resulted in a significant reduction of lignin at cell corners, compound middle lamellae and secondary wall layers of fibres. This study revealed the dynamic developmental changes in the structure of the cell wall during secondary wall development by assembly of cell wall polysaccharides and lignin during differentiation of living fibres in L. leucocephala. These insights are very important to understand the development and functional dynamics of wood fibres.

Glucuronoxylan is the main hemicellulose in the secondary cell wall of angiosperms. Elucidating its molecular structure provides a basis for more accurate plant cell wall models and the utilization of xylan in biorefinery processes. Here, we investigated the spacing of acetyl, glucuronopyranosyl and galactopyranosyl substitutions on Eucalyptus glucuronoxylan using sequential extraction combined with enzymatic hydrolysis and mass spectrometry. We found that the acetyl groups are preferentially spaced with an even pattern and that consecutive acetylation is present as a minor motif. Distinct odd and even patterns of glucuronidation with tight and sparse spacing were observed. Furthermore, the occurrence of consecutive glucuronidation is reported, which adds to the growing body of evidence that this motif is not only present in gymnosperms but also in angiosperms. In addition, the presence of terminal galactopyranosyl units, which can be released by β-galactosidase, altered the digestibility of the glucuronoxylan by GH30 and GH10 xylanase and appeared to be clustered within the polymeric backbone. These findings increase our understanding of the complex structure of glucuronoxylans and its effect on the extractability and biological degradation of Eucalyptus wood.

Overcoming lignocellulose recalcitrance to enzymatic or chemical processing is a prerequisite for biorefinery applications. Expansins and loosenins are non-lytic proteins that could assist reducing this recalcitrance by disrupting the intermolecular contacts between plant cell wall components. Here, immunolocalization with fluorescence and transmission electron microscopy (TEM) were used to study the ability of a Bacillus subtilis expansin-like protein (BsEXLX1), a Phanerochaete carnosa loosenin protein (PcaLOOL12) and a fusion protein of PcaLOOL12 with the carbohydrate-binding module 63 (CBM63) of BsEXLX1 (i.e., PcaLOOL12-CBM63) to bind secondary cell walls (SCW) of aspen fibres, including fresh aspen wood, milled wood fibres (MWF) and MWF subjected to subcritical water extraction. The immunofluorescence labelling of fresh wood samples showed a weak signal for PcaLOOL12 and a strong signal for BsEXLX1 and PcaLOOL12-CBM63, suggesting the importance of CBM63 for protein adsorption to SCW components. TEM analysis after immunogold labelling revealed the presence of BsEXLX1 and PcaLOOL12-CBM63 in all secondary cell wall layers. Pretreatment of wood samples with the proteins reduced the binding of glucomannan- and glucuronoxylan-specific monoclonal antibodies. Similarly, protein adsorption to MWF was higher before subcritical water extraction. Together, these results suggest the adsorption of BsEXLX1 and PcaLOOL12-CBM63 to SCWs was mediated at least in part by their interaction with hemicelluloses. Our study demonstrates that microbial expansin-related proteins can bind to the secondary walls of aspen wood through potential interaction of CBM63 with hemicelluloses.

Microbial expansin-related proteins include fungal loosenins, which have been previously shown to disrupt cellulose networks and enhance the enzymatic conversion of cellulosic substrates. Despite showing beneficial impacts to cellulose processing, detailed characterization of cellulosic materials after loosenin treatment is lacking. In this study, small-angle neutron scattering (SANS) was used to investigate the effects of three recombinantly produced loosenins that originate from Phanerochaete carnosa, PcaLOOL7, PcaLOOL9, and PcaLOOL12, on the organization of holocellulose preparations from Eucalyptus and Spruce wood samples. Whereas the SANS analysis of Spruce holocellulose revealed an increase in inter-microfibril spacing of neighboring cellulose microfibrils following treatment with PcaLOOL12 and to a lesser extent PcaLOOL7, the analysis of Eucalyptus holocellulose revealed a reduction in the ordered arrangement of microfibrils following treatment with PcaLOOL12 and to a lesser extent PcaLOOL9. Parallel SEC-SAXS characterization of PcaLOOL7, PcaLOOL9, and PcaLOOL12 indicated the proteins likely function as monomers; moreover, all appear to retain a flexible disordered N-terminus and folded C-terminal region. The comparatively high impact of PcaLOOL12 motivated its NMR structural characterization, revealing a double-psi β-barrel (DPBB) domain surrounded by three α-helices—the largest nestled against the DPBB core and the other two part of loops extending from the core. The SANS analysis of PcaLOOL action on holocellulose samples confirms their ability to disrupt cellulose fiber networks and suggests a progression from reducing regular order in the microfibril arrangement to increasing inter-microfibril spacing. The most impactful PcaLOOL, PcaLOOL12, was previously observed to be the most highly expressed loosenin in P. carnosa. Its structural characterization herein reveals its stabilization through two disulfide linkages, and an extended N-terminal region distal to a negatively charged and surface accessible polysaccharide binding groove.

Wood of broad-leaf tree species is a valued source of renewable biomass for biorefinery and a target for genetic improvement efforts to reduce its recalcitrance. Glucuronoxylan (GX) plays a key role in recalcitrance through its interactions with cellulose and lignin. To reduce recalcitrance, we modified wood GX by expressing GH10 and GH11 endoxylanases from Aspergillus nidulans in hybrid aspen (Populus tremula L. x tremuloides Michx.) and targeting the enzymes to cell wall. The xylanases reduced tree height, modified cambial activity by increasing phloem and reducing xylem production, and reduced secondary wall deposition. Xylan molecular weight was decreased, and the spacing between acetyl and MeGlcA side chains was reduced in transgenic lines. The transgenic trees produced hypolignified xylem having thin secondary walls and deformed vessels. Glucose yields of enzymatic saccharification without pretreatment almost doubled indicating decreased recalcitrance. The transcriptomics, hormonomics and metabolomics data provided evidence for activation of cytokinin and ethylene signalling pathways, decrease in ABA levels, transcriptional suppression of lignification and a subset of secondary wall biosynthetic program, including xylan glucuronidation and acetylation machinery. Several candidate genes for perception of impairment in xylan integrity were detected. These candidates could provide a new target for uncoupling negative growth effects from reduced recalcitrance. In conclusion, our study supports the hypothesis that xylan modification generates intrinsic signals and evokes novel pathways regulating tree growth and secondary wall biosynthesis. Fungal xylanases expressed in aspen decreased lignin content in the wood of transgenic lines due to downregulation of a subset of lignin biosynthetic genes. This greatly increased saccharification potential. Transgenic plants exhibited altered growth and several signaling and regulatory genes were found affected that could mediate these effects via xylobiose signaling.

BACKGROUND:Microbial expansin-related proteins include fungal loosenins, which have been previously shown to disrupt cellulose networks and enhance the enzymatic conversion of cellulosic substrates. Despite showing beneficial impacts to cellulose processing, detailed characterization of cellulosic materials after loosenin treatment is lacking. In this study, small-angle neutron scattering (SANS) was used to investigate the effects of three recombinantly produced loosenins that originate from Phanerochaete carnosa, PcaLOOL7, PcaLOOL9, and PcaLOOL12, on the organization of holocellulose preparations from Eucalyptus and Spruce wood samples. RESULTS:Whereas the SANS analysis of Spruce holocellulose revealed an increase in interfibril spacing of neighboring cellulose microfibrils following treatment with PcaLOOL12 and to a lesser extent PcaLOOL7, the analysis of Eucalyptus holocellulose revealed a reduction in packing number following treatment with PcaLOOL12 and to a lesser extent PcaLOOL9. Parallel SEC-SAXS characterization of PcaLOOL7, PcaLOOL9, and PcaLOOL12 indicated the proteins likely function as monomers; moreover, all appear to retain a flexible disordered N-terminus and folded C-terminal region. The comparatively high impact of PcaLOOL12 motivated its NMR structural characterization, revealing a double-psi b-barrel (DPBB) domain surrounded by three alpha-helices - the largest nestled against the DPBB core and the other two part of loops extending from the core. CONCLUSIONS:The SANS analysis of PcaLOOL action on holocellulose samples confirms their ability to disrupt cellulose fiber networks and suggests a progression from reducing microfibril packing to increasing interfibril distance. The most impactful PcaLOOL, PcaLOOL12, was previously observed to be the most highly expressed loosenin in P. carnosa. Its structural characterization herein reveals its stabilization through two disulfide linkages, and an extended N-terminal region distal to a negatively charged and surface accessible polysaccharide binding groove.

SummaryWood of broad‐leaf tree species is a valued source of renewable biomass for biorefinery and a target for genetic improvement efforts to reduce its recalcitrance. Glucuronoxylan (GX) plays a key role in recalcitrance through its interactions with cellulose and lignin. To reduce recalcitrance, we modified wood GX by expressing GH10 and GH11 endoxylanases from Aspergillus nidulans in hybrid aspen (Populus tremula L. × tremuloides Michx.) and targeting the enzymes to cell wall. The xylanases reduced tree height, modified cambial activity by increasing phloem and reducing xylem production, and reduced secondary wall deposition. Xylan molecular weight was decreased, and the spacing between acetyl and MeGlcA side chains was reduced in transgenic lines. The transgenic trees produced hypolignified xylem having thin secondary walls and deformed vessels. Glucose yields of enzymatic saccharification without pretreatment almost doubled indicating decreased recalcitrance. The transcriptomics, hormonomics and metabolomics data provided evidence for activation of cytokinin and ethylene signalling pathways, decrease in ABA levels, transcriptional suppression of lignification and a subset of secondary wall biosynthetic program, including xylan glucuronidation and acetylation machinery. Several candidate genes for perception of impairment in xylan integrity were detected. These candidates could provide a new target for uncoupling negative growth effects from reduced recalcitrance. In conclusion, our study supports the hypothesis that xylan modification generates intrinsic signals and evokes novel pathways regulating tree growth and secondary wall biosynthesis.

Xylan is a fundamental structural polysaccharide in plant secondary cell walls and a valuable resource for biorefinery applications. Deciphering the molecular motifs of xylans that mediate their interaction with cellulose and lignin is fundamental to understand the structural integrity of plant cell walls and to design lignocellulosic materials. In the present study, we investigated the pattern of acetylation and glucuronidation substitution in hardwood glucuronoxylan (GX) extracted from aspen wood using subcritical water and alkaline conditions. Enzymatic digestions of GX with β-xylanases from glycosyl hydrolase (GH) families GH10, GH11 and GH30 generated xylo-oligosaccharides with controlled structures amenable for mass spectrometric glycan sequencing. We identified the occurrence of intramolecular motifs in aspen GX with block repeats of even glucuronidation (every 2 xylose units) and consecutive glucuronidation, which are unique features for hardwood xylans. The acetylation pattern of aspen GX shows major domains with evenly-spaced decorations, together with minor stretches of highly acetylated domains. These heterogenous patterns of GX can be correlated with its extractability and with its potential interaction with lignin and cellulose. Our study provides new insights into the molecular structure of xylan in hardwood species, which has fundamental implications for overcoming lignocellulose recalcitrance during biochemical conversion.

Tissue cultures are an important study model for woody plant tissue and can be used to study lignin biosynthesis. The greatest disadvantage of protocols based on extraction of lignin from wood biomass is the almost inevitable alteration of the native structure of lignin. Using a Norway spruce tissue culture with the ability to secrete monolignols into a liquid culture medium, fundamental aspects of lignin have been studied in the past, such as its structure, the enzyme activity related to its polymerization, and its interactions with a secondary cell wall hemicellulose. In this study, parameters that can induce monolignol production and secretion in the tissue culture are investigated via gene expression analysis. The impact of the composition of the solid growth medium, which was in some cases supplemented with xylan, was studied in depth through transcriptomic investigation. We find that the state (i.e. liquid or solid) and the xylan content of the medium can impact gene expression, although microscopic analysis suggests that cellular morphology is consistent. Extracellular lignin was collected from a formulation of liquid medium with the same composition as that used for cellular growth, which was previously presumed to be “non-inducing” of lignin biosynthesis. Chemical analysis of this lignin was performed using nuclear magnetic resonance spectroscopy and size exclusion chromatography, which revealed changes in its structure compared to the polymer produced in the previously developed “inducing” liquid medium. These experiments show that there is still much we do not understand about an oft-used tissue culture system, but show the way to a deeper understanding of the genetic control of lignin biosynthesis. ### Competing Interest Statement The authors have declared no competing interest.

The structural integrity of cell walls in wood is maintained by matrix polysaccharides through chemical interactions among themselves and with both cellulose and lignin. During wood formation, matrix polysaccharides are deposited in the cell walls and subsequently modified by spontaneous and/or enzyme-mediated processes. These processes are highly regulated during successive stages of wood formation, including cell division, expansion and secondary (or further tertiary) cell wall layer deposition. They are also adjusted in response to various endogenous and environmental signals by developing xylem cells, with corresponding modifications of cell wall properties. In addition, there is substantial variation among taxonomic groups of plants in the chemical composition of matrix wood polysaccharides. This chapter discusses these dynamic changes in matrix polymers during wood development and adaptive responses to environmental stresses, the matrix variation among cell types and between hardwoods and softwoods and the functional significance of this dynamic variation for wood cell wall properties.