Plant micro- and nanomechanics: experimental techniques for plant cell-wall analysis

In the last few decades, micro- and nanomechanical methods have become increasingly important analytical techniques to gain deeper insight into the nanostructure and mechanical design of plant cell walls. The objective of this article is to review the most common micro- and nanomechanical approaches that are utilized to study primary and secondary cell walls from a biomechanics perspective. In light of their quite disparate functions, the common and opposing structural features of primary and secondary cell walls are reviewed briefly. A significant part of the article is devoted to an overview of the methodological aspects of the mechanical characterization techniques with a particular focus on new developments and advancements in the field of nanomechanics. This is followed and complemented by a review of numerous studies on the mechanical role of cellulose fibrils and the various matrix components as well as the polymer interactions in the context of primary and secondary cell-wall functio

Properties of chemically and mechanically isolated fibres of spruce (Picea abies [L.] Karst.). Part 1: Structural and chemical characterisation

Single fibres of spruce (Picea abies [L.] Karst.) were isolated both chemically and mechanically from a solid wood sample. Mechanical isolation was carried out using very fine tweezers to peel out fibres, thereby taking advantage of the low shear strength between them. Chemical isolation was achieved using hydrogen peroxide and glacial acetic acid. Fibres were examined with Fourier-transform infrared (FT-IR) microscopy, and field-emission environmental scanning electron microscopy (FE-ESEM) in low-Vacuum mode to compare the isolation techniques with respect to their influence on cell wall structure and polymer assembly. The chemical treatment led to degradation of lignin and hemicelluloses, significantly influencing the cell wall assembly and structure. The cell wall polymers of mechanically isolated fibres remained in their natural constitution. As expected, the peeling process caused separation of cell wall layers. Our examinations indicate that delamination predominately took place at the interface between the secondary cell wall and the compound middle lamella. However, fracture between the S1 and S2 layers was examined as well. With respect to fibre quality, it was of particular importance that transverse crack propagation in the secondary cell walls (S2) was not observe

Micromechanical properties of common yew ( Taxus baccata ) and Norway spruce ( Picea abies ) transition wood fibers subjected to longitudinal tension

The longitudinal modulus of elasticity of common yew is astonishingly low in light of its high raw density. At least this was found for specimens examined at the solid wood level and at the tissue level. However, to reveal if this low axial stiffness is also present at the cellular level, tensile tests were performed on individual yew fibers and on spruce fibers for reference. The results revealed a low stiffness and a high strain to fracture for yew when compared with spruce. This compliant behavior was ascribed to a relatively high microfibril angle of yew measured by X-ray scattering. It can be concluded that the high compliance of yew observed at higher hierarchical levels is obviously controlled by a structural feature present at the cell wall level. In future studies, the biomechanical function of this compliant behavior for the living yew tree would be of particular interes

Plant material features responsible for bamboo's excellent mechanical performance: a comparison of tensile properties of bamboo and spruce at the tissue, fibre and cell wall levels

Background and Aims Bamboo is well known for its fast growth and excellent mechanical performance, but the underlying relationships between its structure and properties are only partially known. Since it lacks secondary thickening, bamboo cannot use adaptive growth in the same way as a tree would in order to modify the geometry of the stem and increase its moment of inertia to cope with bending stresses caused by wind loads. Consequently, mechanical adaptation can only be achieved at the tissue level, and this study aims to examine how this is achieved by comparison with a softwood tree species at the tissue, fibre and cell wall levels. Methods The mechanical properties of single fibres and tissue slices of stems of mature moso bamboo (Phyllostachys pubescens) and spruce (Picea abies) latewood were investigated in microtensile tests. Cell parameters, cellulose microfibril angles and chemical composition were determined using light and electron microscopy, wide-angle X-ray scattering and confocal Raman microscopy. Key Results Pronounced differences in tensile stiffness and strength were found at the tissue and fibre levels, but not at the cell wall level. Thus, under tensile loads, the differing wall structures of bamboo (multilayered) and spruce (sandwich-like) appear to be of minor relevance. Conclusions The superior tensile properties of bamboo fibres and fibre bundles are mainly a result of amplified cell wall formation, leading to a densely packed tissue, rather than being based on specific cell wall properties. The material optimization towards extremely compact fibres with a multi-lamellar cell wall in bamboo might be a result of a plant growth strategy that compensates for the lack of secondary thickening growth at the tissue level, which is not only favourable for the biomechanics of the plant but is also increasingly utilized in terms of engineering products made from bamboo culm

Hydroxyl accessibility in wood cell walls as affected by drying and re-wetting procedures

The first drying of wood cell walls from the native state has sometimes been described as producing irreversible structural changes which reduce the accessibility to water, a phenomenon often referred to as hornification. This study demonstrates that while changes do seem to take place, these are more complex than what has hitherto been described. The accessibility of wood cell wall hydroxyls to deuteration in the form of liquid water was not found to be affected by drying, since vacuum impregnation with liquid water restores the native cell wall accessibility. Contrary to this, hydroxyl accessibility to deuteration by water vapour was found to decrease to different levels depending on the drying conditions. Vacuum drying at 60 °C for 3 days reduced the accessibility more than drying for 1 day at 103 °C without vacuum. Drying for 3 days at 103 °C increased the hydroxyl accessibility compared to 1 day. Moreover, the decrease in hydroxyl accessibility to deuteration by water vapour induced by the first drying could be at least partially erased by subsequent vacuum impregnation with liquid water, indicating reversibility. For the drying of solid, non-degraded wood cell walls the results challenge the often supposed process of hornification, understood as a permanent decrease in hydroxyl accessibility to water.ISSN:1572-882XISSN:0969-023

Significant influence of lignin on axial elastic modulus of poplar wood at low microfibril angles under wet conditions

Wood is extensively used as construction material. Despite accumulating knowledge on its mechanical properties, the contribution of the cell wall matrix polymers to wood mechanics is still not well understood. Previous studies have shown that axial stiffness correlates with lignin content only for cellulose microfibril angles larger than around 20°, while no influence was found for smaller angles. Here, by analyzing the wood of poplar plants with reduced lignin content due to down-regulation of CAFFEOYL SHIKIMATE ESTERASE, we showed that lignin content influences axial stiffness also at smaller angles. Micro-tensile tests of the xylem revealed that axial stiffness was strongly reduced in the low-lignin transgenic lines. Strikingly, microfibril angles were around 15° for both wild type and transgenic poplars, suggesting that cellulose orientation is not responsible for the observed changes in mechanical behavior. Multiple linear regression analysis showed that the decrease in stiffness was almost completely related to the variation in both density and lignin content. We suggest that the influence of lignin content on axial stiffness may gradually increase as a function of the microfibril angle. The obtained results may help setting up comprehensive models of the cell wall for unraveling the individual role of the cell wall matrix polymers

Mineralization of wood by calcium carbonate insertion for improved flame retardancy

Wood can be considered as a highly porous, three-dimensional organic scaffold. It can be mineralized to create hierarchically structured organic-inorganic hybrid materials with novel properties. In the present paper, the precipitation of CaCO3 mineral in Norway spruce and European beech wood has been studied by alternating impregnation with aqueous and alcoholic electrolyte solutions. Microstructural imaging by SEM and confocal Raman microscopy shows the distribution of calcite and vaterite as two CaCO3 polymorphs, which are deposited deep inside the cellular structure of the wood. The confined microenvironment of the wood cell wall seems to favor a formation of vaterite, as visible by XRD and Raman spectroscopy. In view of a practical application, the mineralization of wood opens up ways for sustainable wood-based hybrid materials with a significantly improved fire resistance, as proven via pyrolysis combustion flow calorimetry and cone calorimetry tests. Beyond that, this versatile solute-exchange approach provides an opportunity for the incorporation of a broad range of different mineral phases into wood for novel material property combinations

Biotehnološka razgradnja i molekularni mehanizmi razgradnje drveta pomoću selektivnih gljiva, uzročnika bijele truleži

Microbial mechanisms of lignin degradation may be utilised for solid-state fermentations other than biopulping, during which the selective conversion of lignin is required. The current paper reviews current work into selective lignin conversion, with emphasis on the contributions made by our research group, which consists of researchers from five different laboratories. Three of them cooperate within Wood K plus. The recent research of this group has focussed on fermentations utilising the unique metabolism of selective white-rot fungi to modify wood surfaces during relatively short fermentation times of less than one week and on research into the molecular mechanisms causing these modifications. Lignin degradation by selective fungi (e.g. Ceriporiopsis subvermispora and species of the genus Phlebia) on the wood surfaces was significant after three days. After seven days the overall lignin content of spruce wood shavings was reduced by more than 3.5 %. Lignin loss was accompanied by an increase of extractable substances. To evaluate small changes and to trace the fungal modification processes, Fourier transform infrared spectroscopic (FTIR) techniques and electron paramagnetic resonance (EPR) spectroscopy were applied and adapted. The spectra recorded in the near infrared region (FT-NIR) turned out to be very useful for kinetic studies of the biopulping/biomodification processes and a good method to evaluate the capabilities of fungi to modify wood surfaces within this short period.Mikrobiološki mehanizmi razgradnje lignina, osim biološke proizvodnje pulpe tijekom koje dolazi do selektivne pretvorbe lignina, mogu se primijeniti tijekom fermentacije na krutoj podlozi. U ovom je revijalnom prikazu dan pregled istraživanja selektivne pretvorbe lignina, a osobito rad znanstvenoga tima, koji se sastoji od istraživača iz pet raznih laboratorija. Troje istraživaa surađuju u centru Wood K plus. Istraživanja te skupine bila su usmjerena na fermentaciju primjenom jedinstvenog metabolizma selektivne gljive, uzročnika bijele truleži, u razgradnji površine drveta tijekom relativno kratkog vremena fermentacije (manje od tjedan dana) i na istraživanje molekularnih mehanizama koji uzrokuju te promjene. Razgradnja lignina s pomoću selektivnih gljiva (npr. Ceriporiopsis subvermispora i vrste roda Phlebia) na površini drveta bila je značajna nakon tri dana. Nakon sedam dana ukupni udjel lignina u piljevini drva smreke smanjen je za više od 3,5 %. Gubitak lignina praćen je povećanjem količine ekstraktibilnih tvari. Da bi se pratio proces modifikacije s pomoću gljiva, primijenjene su prilagođene metode Fourier transformacijske infracrvene spektroskopije (FTIR) i elektronske paramagnetske rezonancije (EPR). Spektar snimljen blizu infracrvenog područja (FT-NIR) bio je vrlo koristan za istraživanje kinetike biološke proizvodnje pulpe odnosno procesa biomodifikacije i dobra je metoda za procjenu sposobnosti gljiva da u vrlo kratkom vremenskom roku razgrađuju površinu drveta

Even Visually Intact Cell Walls in Waterlogged Archaeological Wood Are Chemically Deteriorated and Mechanically Fragile: A Case of a 170 Year-Old Shipwreck

Structural and chemical deterioration and its impact on cell wall mechanics were investigated for visually intact cell walls (VICWs) in waterlogged archaeological wood (WAW). Cell wall mechanical properties were examined by nanoindentation without prior embedding. WAW showed more than 25% decrease of both hardness and elastic modulus. Changes of cell wall composition, cellulose crystallite structure and porosity were investigated by ATR-FTIR imaging, Raman imaging, wet chemistry, 13C-solid state NMR, pyrolysis-GC/MS, wide angle X-ray scattering, and N2 nitrogen adsorption. VICWs in WAW possessed a cleavage of carboxyl in side chains of xylan, a serious loss of polysaccharides, and a partial breakage of β-O-4 interlinks in lignin. This was accompanied by a higher amount of mesopores in cell walls. Even VICWs in WAW were severely deteriorated at the nanoscale with impact on mechanics, which has strong implications for the conservation of archaeological shipwrecks

Even Visually Intact Cell Walls in Waterlogged Archaeological Wood Are Chemically Deteriorated and Mechanically Fragile : A Case of a 170 Year-Old Shipwreck

Structural and chemical deterioration and its impact on cell wall mechanics were investigated for visually intact cell walls (VICWs) in waterlogged archaeological wood (WAW). Cell wall mechanical properties were examined by nanoindentation without prior embedding. WAW showed more than 25% decrease of both hardness and elastic modulus. Changes of cell wall composition, cellulose crystallite structure and porosity were investigated by ATR-FTIR imaging, Raman imaging, wet chemistry, C-13-solid state NMR, pyrolysis-GC/MS, wide angle X-ray scattering, and N-2 nitrogen adsorption. VICWs in WAW possessed a cleavage of carboxyl in side chains of xylan, a serious loss of polysaccharides, and a partial breakage of beta -O-4 interlinks in lignin. This was accompanied by a higher amount of mesopores in cell walls. Even VICWs in WAW were severely deteriorated at the nanoscale with impact on mechanics, which has strong implications for the conservation of archaeological shipwrecks.Peer reviewe