Why Do Bamboo Parenchyma Cells Show Higher Nanofibrillation Efficiency than Fibers: An Investigation on Their Hierarchical Cell Wall Structure.

The cell walls of parenchyma cells and fibers in bamboo are both highly lignified with secondary thickening. However, the former were found to have much higher nanofibrillation efficiency than fibers via both protocols of ultrasonication and high pressure homogenization. To elucidate the inherent mechanism, detailed comparisons of chemical composition, cell morphology, cell wall density, pore structures, and structural organization of cell wall polymers were performed on native and pretreated cell walls of both parenchyma cells and fibers. Chemical compositional analysis showed that fibers have much higher cellulose (49.8% to 35.5%) but lower xylan content (21.1% to 36.2%) than parenchyma, while their lignin contents were similar (24.9% vs 22.9%). Polarized FTIR further revealed clear differences in the structural organization of polymers between the two types of cells, with all the polymers of fibers being more orderly assembled than those of parenchyma cells. The compact arrangement of polymers in the fibers was also supported by the much higher cell wall density (1.52 vs 1.28 g/cm3) and lower porosity (0.007 vs 0.013 cc/g after chemical pretreatments), as compared to the parenchyma cells. The study provides evidence that the anatomical characteristics of huge cavity-wall ratio, higher cell wall porosity, and less ordered arrangement of cell wall matrix polymers (mainly lignin) in parenchyma cells contribute to their higher nanofibrillation efficiency compared to fibers.

Cell wall polymer distribution in bamboo visualized with in situ imaging FTIR.

To better understand the high recalcitrance of bamboo during bioconversion, the fine spatial distribution of polymers in bamboo was studied with Imaging FTIR microscopy under both transmission and ATR modes, combined with PCA data processing. The results demonstrated that lignin, xylan and hydroxycinnamic acid (HCA) were more concentrated in the fibers near the xylem conduit, while cellulose was evenly distributed across the whole fiber sheath. PCA processing produced a clear separation between bamboo fibers and parenchyma cells, indicating that the parenchyma cells contains more pectin and HCA than fibers. It also demonstrated that cellulose, xylan and S-lignin were concentrated most heavily in bamboo fiber secondary cell walls, while G-lignin, pectin and HCA were found more in the compound middle lamella. The revealed information regarding polymer distribution is of great significance for better understanding of the inherent design mechanism of plant cell wall and its efficient utilization.

Molecular origin of mechano-sorptive creep in cellulosic fibres.

Mechano-sorptive creep, i.e. the increased rate of creep that occurs during changing relative humidity, when loading paper or wood, is a phenomenon still not fully understood. This phenomenon was here investigated by examining the changes occurring at the molecular level utilising FTIR spectroscopy. By subjecting the paper to deuterated water, the changes involving both the crystalline hydroxyls as well as those in accessible regions could be examined. During loading, the cellulose molecular chains are stretched taking the load. In addition, during mechano-sorptive creep a further exchange from OH to OD groups occurred. This was interpreted as caused by slippage between cellulose fibrils allowing previously non-accessible hydroxyls to become available for deuterium exchange. Thus, the loosening of the structure, during the changing moisture conditions, is interpreted as what has led to the increased creep and the possibility for new areas of cellulose fibril/fibril aggregates to be exposed to the deuterium exchange.

Structural organization of the cell wall polymers in compression wood as revealed by FTIR microspectroscopy.

MAIN CONCLUSION: Glucomannan was more strongly oriented, in line with the orientation of cellulose, than the xylan in both compression wood and normal wood of Chinese fir. Lignin in compression wood was somewhat more oriented in the direction of the cellulose microfibrils than in normal wood. The structural organization in compression wood (CW) is quite different from that in normal wood (NW). To shed more light on the structural organization of the polymers in plant cell walls, Fourier Transform Infrared (FTIR) microscopy in transmission mode has been used to compare the S2-dominated mean orientation of wood polymers in CW with that in NW from Chinese fir (Cunninghamia lanceolata). Polarized FTIR measurements revealed that in both CW and NW samples, glucomannan and xylan showed a parallel orientation with respect to the cellulose microfibrils. In both wood samples, the glucomannan showed a much greater degree of orientation than the xylan, indicating that the glucomannan has established a stronger interaction with cellulose than xylan. For the lignin, the absorption peak also indicated an orientation along the direction of the cellulose microfibrils, but this orientation was more pronounced in CW than in NW, indicating that the lignin is affected by the orientation of the cellulose microfibrils more strongly in CW than it is in NW.

On the origin of sorption hysteresis in cellulosic materials.

Moisture sorption and moisture sorption hysteresis of carbohydrates are phenomena which affect the utilisation of products made thereof. Although extensively studied, there is still no consensus regarding the mechanisms behind sorption hysteresis. Attempts have been made to link the behaviour to molecular properties, in particular to softening properties, and the moisture sorption hysteresis has therefore here been investigated by modifying cellulosic fibres to affect their softening properties. The results show that the moisture sorption hysteresis diminishes with decreasing softening temperature, and was even completely absent at the higher degrees of modification. The moisture sorption characteristics also changed from a type II sorption to a more type III sorption behaviour, a feature more prominent the higher the degree of modification and the higher the temperature. For the highest degree of modification studied the sorption characteristics changed from sorbing less water the higher the temperature to sorbing more water with increasing temperature.

Non-exchanging hydroxyl groups on the surface of cellulose fibrils: The role of interaction with water.

The interaction of water with cellulose stages many unresolved questions. Here 2H MAS NMR and IR spectra recorded under carefully selected conditions in 1H-2H exchanged, and re-exchanged, cellulose samples are presented. It is shown here, by a quantitative and robust approach, that only two of the three available hydroxyl groups on the surface of cellulose fibrils are exchanging their hydrogen with the surrounding water molecules. This finding is additionally verified and explained by MD simulations which demonstrate that the 1HO(2) and 1HO(6) hydroxyl groups of the constituting glucose units act as hydrogen-bond donors to water, while the 1HO(3) groups behave exclusively as hydrogen-bond acceptors from water and donate hydrogen to their intra-chain neighbors O(5). We conclude that such a behavior makes the latter hydroxyl group unreactive to hydrogen exchange with water.

Changes of wood cell walls in response to hygro-mechanical steam treatment.

The effects of compression combined with steam treatment (CS-treatment), i.e. a hygro-mechanical steam treatment on Spruce wood were studied on a cell-structure level to understand the chemical and physical changes of the secondary cell wall occurring under such conditions. Specially, imaging FT-IR microscopy, nanoindentation and dynamic vapour absorption were used to track changes in the chemical structure, in micromechanical and hygroscopic properties. It was shown that CS-treatment resulted in different changes in morphological, chemical and physical properties of the cell wall, in comparison with those under pure steam treatment. After CS-treatment, the cellular structure displayed significant deformations, and the biopolymer components, e.g. hemicellulose and lignin, were degraded, resulting in decreased hygroscopicity and increased mechanical properties of the wood compared to both untreated and steam treated wood. Moreover, CS-treatment resulted in a higher degree of degradation especially in earlywood compared to a more uniform behaviour of wood treated only by steam.

Deposition and organisation of cell wall polymers during maturation of poplar tension wood by FTIR microspectroscopy.

To advance our understanding of the formation of tension wood, we investigated the macromolecular arrangement in cell walls by Fourier transform infrared microspectroscopy (FTIR) during maturation of tension wood in poplar (Populus tremula x P. alba, clone INRA 717-1B4). The relation between changes in composition and the deposition of the G-layer in tension wood was analysed. Polarised FTIR measurements indicated that in tension wood, already before G-layer formation, a more ordered structure of carbohydrates at an angle more parallel to the fibre axis exists. This was clearly different from the behaviour of opposite wood. With the formation of the S2 layer in opposite wood and the G-layer in tension wood, the orientation signals from the amorphous carbohydrates like hemicelluloses and pectins were different between opposite wood and tension wood. For tension wood, the orientation for these bands remains the same all along the cell wall maturation process, probably reflecting a continued deposition of xyloglucan or xylan, with an orientation different to that in the S2 wall throughout the whole process. In tension wood, the lignin was more highly oriented in the S2 layer than in opposite wood.

Xylan as limiting factor in enzymatic hydrolysis of nanocellulose.

The role of xylan as a limiting factor in the enzymatic hydrolysis of cellulose was studied by hydrolysing nanocellulose samples prepared by mechanical fibrillation of birch pulp with varying xylan content. Analyzing the nanocelluloses and their hydrolysis residues with dynamic FT-IR spectroscopy revealed that a certain fraction of xylan remained tightly attached to cellulose fibrils despite partial hydrolysis of xylan with xylanase prior to pulp fibrillation and that this fraction remained in the structure during the hydrolysis of nanocellulose with cellulase mixture as well. Thus, a loosely bound fraction of xylan was predicted to have been more likely removed by purified xylanase. The presence of loosely bound xylan seemed to limit the hydrolysis of crystalline cellulose, indicated by an increase in cellulose crystallinity and by preserved crystal width measured with wide-angle X-ray scattering. Removing loosely bound xylan led to a proportional hydrolysis of xylan and cellulose with the cellulase mixture.

Ultra-structural organisation of cell wall polymers in normal and tension wood of aspen revealed by polarisation FTIR microspectroscopy.

Polarisation Fourier transform infra-red (FTIR) microspectroscopy was used to characterize the organisation and orientation of wood polymers in normal wood and tension wood from hybrid aspen (Populus tremula × Populus tremuloides). It is shown that both xylan and lignin in normal wood are highly oriented in the fibre wall. Their orientation is parallel with the cellulose microfibrils and hence in the direction of the fibre axis. In tension wood a similar orientation of lignin was found. However, in tension wood absorption peaks normally assigned to xylan exhibited a 90° change in the orientation dependence of the vibrations as compared with normal wood. The molecular origin of these vibrations are not known, but they are abundant enough to mask the orientation dependence of the xylan signal from the S2 layer in tension wood and could possibly come from other pentose sugars present in, or associated with, the gelatinous layer of tension wood fibres.

Effect of steam treatment on the properties of wood cell walls.

Steam treatment is a hygrothermal method of potential industrial significance for improving the dimensional stability and durability of wood materials. The steaming results in different chemical and micromechanical changes in the nanostructured biocomposite that comprise a wood cell wall. In this study, spruce wood ( Picea abies Karst.) that had been subjected to high-temperature steaming up to 180 °C was examined, using imaging Fourier Transform Infrared (FT-IR) microscopy and nanoindentation to track changes in the chemical structure and the micromechanical properties of the secondary cell wall. Similar changes in the chemical components, due to the steam treatment, were found in earlywood and latewood. A progressive degradation of the carbonyl groups in the glucuronic acid unit of xylan and a loss of mannose units in the glucomannan backbone, that is, a degradation of glucomannan, together with a loss of the C═O group linked to the aromatic skeleton in lignin, was found. The development of the hygroscopic and micromechanical properties that occurred with an elevation in the steam temperature correlated well with this pattern of degradation in the constituents in the biocomposite matrix in the cell wall (hemicellulose and lignin).

Localisation and characterisation of incipient brown-rot decay within spruce wood cell walls using FT-IR imaging microscopy.

Spruce wood that had been degraded by brown-rot fungi (Gloeophyllum trabeum or Poria placenta) exhibiting mass losses up to 16% was investigated by transmission Fourier transform infrared (FT-IR) imaging microscopy. Here the first work on the application of FT-IR imaging microscopy and multivariate image analysis of fungal degraded wood is presented and the first report on the spatial distribution of polysaccharide degradation during incipient brown-rot of wood. Brown-rot starts to become significant in the outer cell wall regions (middle lamellae, primary cell walls, and the outer layer of the secondary cell wall S1). This pattern was detected even in a sample with non-detectable mass loss. Most significant during incipient decay was the cleavage of glycosidic bonds, i.e. depolymerisation of wood polysaccharides and the degradation of pectic substances. Accordingly, intramolecular hydrogen bonding within cellulose was reduced, while the presence of phenolic groups increased.

Ultrastructure and mechanical properties of populus wood with reduced lignin content caused by transgenic down-regulation of cinnamate 4-hydroxylase.

Several key enzymes in lignin biosynthesis of Populus have been down-regulated by transgenic approaches to investigate their role in wood lignification and to explore their potential for lignin modification. Cinnamate 4-hydroxylase is an enzyme in the early phenylpropanoid pathway that has not yet been functionally analyzed in Populus . This study shows that down-regulation of cinnamate 4-hydroxylase reduced Klason lignin content by 30% with no significant change in syringyl to guaiacyl ratio. The lignin reduction resulted in ultrastructural differences of the wood and a 10% decrease in wood density. Mechanical properties investigated by tensile tests and dynamic mechanical analysis showed a decrease in stiffness, which could be explained by the lower density. The study demonstrates that a large modification in lignin content only has minor influences on tensile properties of wood in its axial direction and highlights the usefulness of wood modified beyond its natural variation by transgene technology in exploring the impact of wood biopolymer composition and ultrastructure on its material properties.

Pore and matrix distribution in the fiber wall revealed by atomic force microscopy and image analysis.

A method for the ultrastructural investigation of fiber cross-sections based on atomic force microscopy in combination with image analysis is presented. A uniform distribution of pores across the matrix material within the fiber wall was revealed by impregnation of pulp fibers with poly(ethylene glycol). The effects of chemical and mechanical processing on the pore and matrix structure and on the arrangement of the cellulose fibril aggregates were investigated. During chemical processing, changes in the fiber ultrastructure occur: a broadening of the pore and matrix lamella widths in combination with a reduction in their number and an enlargement of the cellulose fibril aggregates. It was found that pores formed during pulping are evenly distributed across the fiber wall in the transverse direction. In contrast, refining increases the pore and matrix lamella width in the fiber wall closest to the middle lamella an effect which gradually decrease in size toward the lumen side.

Micromechanical understanding of the cell-wall structure.

For improving properties of pulp fibres, a better understanding of the relationships between its macroscopic mechanical properties, fibre ultrastructure, and properties of the wood polymers is important. This paper discusses such relations between elastic properties of fibres, their matrix structure and the wood polymer elastic constants. It is argued that an orientation of all of the wood polymers in the direction of the cellulose microfibrils is most likely. The elastic longitudinal modulus of cellulose is well described by the value of 134 GPa dominating the longitudinal fibre properties. In the transverse direction the amorphous polymers play a more important role.

Characterization of the crystalline structure of cellulose using static and dynamic FT-IR spectroscopy.

The cellulose structure is a factor of major importance for the strength properties of wood pulp fibers. The ability to characterize small differences in the crystalline structures of cellulose from fibers of different origins is thus highly important. In this work, dynamic FT-IR spectroscopy has been further explored as a method sensitive to cellulose structure variations. Using a model system of two different celluloses, the relation between spectral information and the relative cellulose Ialpha content was investigated. This relation was then used to determine the relative cellulose Ialpha content in different pulps. The estimated cellulose I allomorph compositions were found to be reasonable for both unbleached and bleached chemical pulps. In addition, it was found that the dynamic FT-IR spectroscopy technique had the potential to indicate possible correlation field splitting peaks of cellulose Ibeta.

The association of water to cellulose and hemicellulose in paper examined by FTIR spectroscopy.

The nature of water sorption to different materials has always been a complex matter to address, partly due to the different possibilities of hydrogen-bond formation. For cellulosic materials this is extremely important for its product performance. In order to gain a deeper understanding of the moisture adsorption mechanisms of cellulose and hemicelluloses, the molecular interaction between moisture and paper and between moisture and some wood polymers was studied using FTIR spectroscopy under stable humid conditions. It was found that all the moisture-sorbing sites adsorbed moisture to the same relative degree, and that the rate of adsorption was the same for all these sites. It was also noticed that the moisture is adsorbed in the form of clusters. A direct relationship was found between the moisture weight gain and the increase in the absorbance peaks for humidities up to 50% relative humidity after which the moisture gain increased faster, a fact that still remains to be explained.

Load distribution in native cellulose.

The properties of cellulose materials are dependent on interactions between and within the cellulose chains. To investigate the deformation behavior of cellulose and its relation to molecular straining, sheets with fibers oriented preferably in one direction were studied by dynamic FT-IR spectroscopy. Celluloses with different origins (spruce pulp, Cladophora cellulose, cotton linters) were used. The sheets were stretched sinusoidally at low strains and small amplitudes while being irradiated with polarized infrared radiation. The cellulose fibers showed mainly an elastic response. The cellulose fibers showed mainly an elastic response. The glucose rings and the C-O-C bridges connecting adjacent rings, as well as the O(3)H.O(5) intramolecular hydrogen bonds are the components mainly deformed under stress, whereas the O(2)H.O(6) intramolecular hydrogen bonds play a minor role. The load distribution was also found to be different in the different allomorphic forms of cellulose I, namely, I(alpha) and I(beta).