Ingo Burgert

Disrupting Two Arabidopsis thaliana Xylosyltransferase Genes Results in Plants Deficient in Xyloglucan, a Major Primary Cell Wall Component

Xyloglucans are the main hemicellulosic polysaccharides found in the primary cell walls of dicots and nongraminaceous monocots, where they are thought to interact with cellulose to form a three-dimensional network that functions as the principal load-bearing structure of the primary cell wall. To determine whether two Arabidopsis thaliana genes that encode xylosyltransferases, XXT1 and XXT2, are involved in xyloglucan biosynthesis in vivo and to determine how the plant cell wall is affected by the lack of expression of XXT1, XXT2, or both, we isolated and characterized xxt1 and xxt2 single and xxt1 xxt2 double T-DNA insertion mutants. Although the xxt1 and xxt2 mutants did not have a gross morphological phenotype, they did have a slight decrease in xyloglucan content and showed slightly altered distribution patterns for xyloglucan epitopes. More interestingly, the xxt1 xxt2 double mutant had aberrant root hairs and lacked detectable xyloglucan. The reduction of xyloglucan in the xxt2 mutant and the lack of detectable xyloglucan in the xxt1 xxt2 double mutant resulted in significant changes in the mechanical properties of these plants. We conclude that XXT1 and XXT2 encode xylosyltransferases that are required for xyloglucan biosynthesis. Moreover, the lack of detectable xyloglucan in the xxt1 xxt2 double mutant challenges conventional models of the plant primary cell wall.

Actuation systems in plants as prototypes for bioinspired devices

Plants have evolved a multitude of mechanisms to actuate organ movement. The osmotic influx and efflux of water in living cells can cause a rapid movement of organs in a predetermined direction. Even dead tissue can be actuated by a swelling or drying of the plant cell walls. The deformation of the organ is controlled at different levels of tissue hierarchy by geometrical constraints at the micrometre level (e.g. cell shape and size) and cell wall polymer composition at the nanoscale (e.g. cellulose fibril orientation). This paper reviews different mechanisms of organ movement in plants and highlights recent research in the field. Particular attention is paid to systems that are activated without any metabolism. The design principles of such systems may be particularly useful for a biomimetic translation into active technical composites and moving devices.

Cell wall features with regard to mechanical performance. A review COST Action E35 2004-2008: Wood machining - micromechanics and fracture

Abstract The mechanical performance of wood and wood products is highly dependent on the structural arrangement and properties of the polymers within the fibre cell wall. To improve utilisation and manufacture of wood materials, there is an increasing need for a more detailed knowledge regarding structure/property relations at the micro- or nanostructural level. In this article, recent developments regarding our understanding of the wood cell wall structure and its mechanical performance are summarised. The new results are interpreted in relation to property performances of wood fibres and wood tissues. Suggestions are made for future requirements for research in this field.

Exploring the micromechanical design of plant cell walls

Plants are hierarchically organized in a way that their macroscopic properties emerge from their micro- and nanostructural level. Hence, micromechanical investigations, which focus on the mechanical design of plant cell walls, are well suited for elucidating the details of the relationship between plant form and function. However, due to the complex nature of primary and secondary cell walls, micromechanical tests on the entire structure cannot provide exact values for polymer properties but must be targeted at the general mechanisms of cell wall deformation and polymer interaction. The success of micromechanical examinations depends on well-considered specimen selection and/or sample pretreatment as well as appropriate experimental setups. Making use of structural differences by taking advantage of the natural variability in plant tissue and cell structure, adaptation strategies can be analyzed at the micro- and nanoscale. Targeted genetic and enzymatic treatments can be utilized to specifically modify individual polymers without degrading the structural integrity of the cell wall. The mechanical properties of such artificial systems reveal the functional roles of individual polymers for a better understanding of the mechanical interactions within the cell wall assembly. In terms of testing methodology, in situ methods that combine micromechanical testing with structural and chemical analyses are particularly well suited for the study of the basic structure-property relationships in plant design. The micromechanical approaches reviewed here are not exhaustive, but they do provide a reasonably comprehensive overview of the methodology with which the general mechanisms underlying the functionality of plant micro- and nanostructure can be explored without destroying the entire cell wall.

On the role of interface polymers for the mechanics of natural polymeric composites

Research on the deformation mechanisms of tendons and wood has shown that these tissues deform mostly by shearing of a soft matrix between stiff fibres. For this type of composite to be both strong and tough, tight binding between matrix and fibres is required. Recent results suggest that Nature may have evolved special interface polymers, capable both of binding to the fibres and of forming a matrix. Proteoglycans could play this role in tendons by binding to collagen fibrils with their protein-like ends and by forming an aqueous matrix with their sugar-like ends. Hemicelluloses could play a similar role in the plant cell wall, as they are binding to cellulose fibrils and forming aqueous networks between them. This observation suggests that new biomimetic composites might be developed on the basis of amphiphilic polymers capable of binding to stiff fibres and of forming a gel-like matrix around them.

Plants control the properties and actuation of their organs through the orientation of cellulose fibrils in their cell walls

Plants use the orientation of cellulose microfibrils to create cell walls with anisotropic properties related to specific functions. This enables organisms to control the shape and size of cells during growth, to adjust the mechanical performance of tissues, and to perform bending movements of organs. We review the key function of cellulose orientation in defining structural-functional relationships in cell walls from a biomechanics perspective, and illustrate this by examples mainly from our own work. First, primary cell-wall expansion largely depends on the organization of cellulose microfibrils in newly deposited tissue and model calculations allow an estimate of how their passive re-orientation may influence the growth of cells. Moreover, mechanical properties of secondary cell walls depend to a large extent on the orientation of cellulose fibrils and we discuss strategies whereby plants utilize this interrelationship for adaptation. Lastly, we address the question of how plants regulate complex organ movements by designing appropriate supramolecular architectures at the level of the cell wall. Several examples, from trees to grasses, show that the cellulose architecture in the cell wall may be used to direct the swelling or shrinking of cell walls and thereby generate internal growth stress or movement of organs.

In situ FT-IR microscopic study on enzymatic treatment of poplar wood cross-sections

The feasibility of Fourier transform infrared (FT-IR) microscopy to monitor in situ the enzymatic degradation of wood was investigated. Cross-sections of poplar wood were treated with cellulase Onozuka RS within a custom-built fluidic cell. Light-optical micrographs and FT-IR spectra were acquired in situ from normal and tension wood fibers. Light-optical micrographs showed almost complete removal of the gelatinous (G) layer in tension wood. No structural and spectral changes were observed in the lignified cell walls. The accessibility of cellulose within the lignified cell wall was found to be the main limiting factor, whereas the depletion of the enzyme due to lignin adsorption could be ruled out. The fast, selective hydrolysis of the crystalline cellulose in the G-layer, even at room temperature, might be explained by the gel-like structure and the highly porous surface. Young plantation grown hardwood trees with a high proportion of G-fibers thus represent an interesting resource for bioconversion to fermentable sugars in the process to bioethanol.

CHITINASE-LIKE1/POM-POM1 and Its Homolog CTL2 Are Glucan-Interacting Proteins Important for Cellulose Biosynthesis in Arabidopsis

Cell wall and cellulose structure is imperative for proper cell elongation and, consequently, the architecture of plants, but components regulating cellulose structure are still elusive. This article shows that the secreted CTL1/POM1 and its close homolog CTL2 interact with glucan-based polymers and influence cellulose crystallinity and cell expansion. Plant cells are encased by a cellulose-containing wall that is essential for plant morphogenesis. Cellulose consists of β-1,4-linked glucan chains assembled into paracrystalline microfibrils that are synthesized by plasma membrane–located cellulose synthase (CESA) complexes. Associations with hemicelluloses are important for microfibril spacing and for maintaining cell wall tensile strength. Several components associated with cellulose synthesis have been identified; however, the biological functions for many of them remain elusive. We show that the chitinase-like (CTL) proteins, CTL1/POM1 and CTL2, are functionally equivalent, affect cellulose biosynthesis, and are likely to play a key role in establishing interactions between cellulose microfibrils and hemicelluloses. CTL1/POM1 coincided with CESAs in the endomembrane system and was secreted to the apoplast. The movement of CESAs was compromised in ctl1/pom1 mutant seedlings, and the cellulose content and xyloglucan structures were altered. X-ray analysis revealed reduced crystalline cellulose content in ctl1 ctl2 double mutants, suggesting that the CTLs cooperatively affect assembly of the glucan chains, which may affect interactions between hemicelluloses and cellulose. Consistent with this hypothesis, both CTLs bound glucan-based polymers in vitro. We propose that the apoplastic CTLs regulate cellulose assembly and interaction with hemicelluloses via binding to emerging cellulose microfibrils.

Stress generation in the tension wood of poplar is based on the lateral swelling power of the G-layer

The mechanism of active stress generation in tension wood is still not fully understood. To characterize the functional interdependency between the G-layer and the secondary cell wall, nanostructural characterization and mechanical tests were performed on native tension wood tissues of poplar (Populus nigra x Populus deltoids) and on tissues in which the G-layer was removed by an enzymatic treatment. In addition to the well-known axial orientation of the cellulose fibrils in the G-layer, it was shown that the microfibril angle of the S2-layer was very large (about 36 degrees). The removal of the G-layer resulted in an axial extension and a tangential contraction of the tissues. The tensile stress-strain curves of native tension wood slices showed a jagged appearance after yield that could not be seen in the enzyme-treated samples. The behaviour of the native tissue was modelled by assuming that cells deform elastically up to a critical strain at which the G-layer slips, causing a drop in stress. The results suggest that tensile stresses in poplar are generated in the living plant by a lateral swelling of the G-layer which forces the surrounding secondary cell wall to contract in the axial direction.

Tensile and compressive stresses in tracheids are induced by swelling based on geometrical constraints of the wood cell

Plants are able to pre-stress their tissues in order to actuate their organs. Here, we demonstrate with two tissue types of the secondary xylem of conifers (normal wood and compression wood of spruce (Picea abies)) that either tensile or compressive stresses can develop in the longitudinal direction during the swelling of the cell wall. This dramatic difference appears to be due mostly to differences in cell geometry and cellulose fibril orientation. A mechanical model was developed to demonstrate swelling experiments with the help of sodium iodide experiments. The reversal of longitudinal extension can be predicted, based on the orientation of the (nearly inextensible) cellulose fibrils and the shape of the cell.