Cell Wall Elasticity Research Articles

Anisotropic Mechanics of Cellular Substrate Under Finite Deformation

The use of cellular substrates for stretchable electronics minimizes not only disruptions to the natural diffusive or convective flow of bio-fluids, but also the constraints on the natural motion of the skin. The existing analytic constitutive models for the equivalent medium of the cellular substrate under finite stretching are only applicable for stretching along the cell walls. This paper aims at establishing an analytic constitutive model for the anisotropic equivalent medium of the cellular substrate under finite stretching along any direction. The model gives the nonlinear stress–strain curves of the cellular substrate that agree very well with the finite element analysis (FEA) without any parameter fitting. For the applied strain <10%, the stress–strain curves are the same for different directions of stretching, but their differences become significant as the applied strain increases, displaying the deformation-induced anisotropy. Comparison of the results for linear and nonlinear elastic cell walls clearly suggests that the nonlinear stress–strain curves of the cellular substrate mainly result from the finite rotation of cell walls.

Controls on the distribution and resilience ofQuercus garryana: ecophysiological evidence of oak's water‐limitation tolerance

AbstractThe composition of forests in Western North America is changing. The decline in the shade‐intolerant Oregon white oak (Quercus garryana var. garryana) is attributed to increased competition with the tall‐growing Douglas fir (Pseudotsuga menziesii var. menziesii) as a result of widespread fire exclusion. In a warmer, drier future, both species will experience increased water stress, and their distribution will depend on ecophysiological adaptations to water limitation, of which little is known forQ. garryana. Here, we report a suite of new ecophysiological observations to better understand the oak's water‐limitation tolerance, in order to predict its fate in a changing climate. Our study site in the Central Belt of the Franciscan Formation in the Northern California Coast Ranges receives almost no rain in the leaf‐on growing season and has limited subsurface water storage capacity. In spite of low pre‐dawn water potentials (Ψ; to below −3.0 MPa), mature trees maintained high rates of sapflow. Sapflow continued due to a high Ψ gradient (≈1.6 MPa on average) at mid‐day between shoots and the subsurface (inferred from pre‐dawn measurements) throughout the dry season. Depletion and recharge of stored water in stem tissue and leaves helped to sustain transpiration. Leaves experienced low Ψ (below −4 MPa), and declining hydraulic conductance yet remained functional. Pressure–volume curve analyses revealed that the maintenance of positive turgor pressures in leaves at low Ψ may be attributable to dynamic adjustment due to changes in cell wall elasticity. The turgor loss point may be of limited use in delineating ecophysiological limits to growth and reproduction, as transpiration and apparently normal physiological behavior continued after pre‐dawn water potentials declined below turgor loss limits inferred from rehydrated leaves. These findings indicate thatQ. garryanais a water‐limitation‐tolerant tree species that maintains hydraulic function as subsurface water supply and atmospheric demand conditions exceed the ranges at whichP. menziesiican operate. These observations can be used to explainQ. garryana's extant species range and anticipate its likely resilience in a warmer climate.

Growth patterns of Phragmites karka under saline conditions depend on the bulk elastic modulus

Salt stress is known to hamper steady-state water flow, which may reduce plant growth. This research was aimed to study the roles of leaf turgor, osmotic adjustment and cell wall elasticity under saline conditions which may reduce biomass production in Phragmites karka (Retz.) Trin, ex. Steud. (a marsh grass). Plants were grown in 0, 100 and 300 mM NaCl and harvested on 3, 7, 15 and 30 days to observe periodic changes in growth and water relations. Leaf number, relative growth rate, and relative elongation rates were higher in the non-saline control than in the plants grown under saline conditions. Plants showed a rapid decline in leaf growth rate (7–15 days) in 300 mM NaCl compared with a delayed response (15–30 days) in 100 mM NaCl. Leaf water potential decreased with increases in salinity after the third day of exposure whereas osmotic potential decreased after the fifteenth day. Low leaf turgor (Ψp) on the third day indicated an initial phase of osmotic stress under saline conditions. Plants maintained higher Ψp in 0 and 100 mM than in 300 mM NaCl. Differences between mid-day and pre-dawn water potential and water saturation deficit were higher in 300 mM NaCl than with other treatments. Water potential and hydraulic capacitance at turgor loss point decreased whereas bulk elastic modulus increased in 300 mM NaCl. Maintenance of turgor and growth at 100 mM NaCl could be related to efficient osmotic adjustment (use of K+ and Cl–), higher WUEi, and lower bulk elasticity whereas poor growth at 300 mM NaCl may have been a consequence of low turgor, decreased cell hydraulic capacitance and higher bulk elastic modulus.

Forces that shape fission yeast cells.

One of the major challenges of modern cell biology is to understand how cells are assembled from nanoscale components into micrometer-scale entities with a specific size and shape. Here I describe how our quest to understand the morphogenesis of the fission yeast Schizosaccharomyces pombe drove us to investigate cellular mechanics. These studies build on the view that cell shape arises from the physical properties of an elastic cell wall inflated by internal turgor pressure. Consideration of cellular mechanics provides new insights into not only mechanisms responsible for cell-shape determination and growth, but also cellular processes such as cytokinesis and endocytosis. Studies in yeast can help to illuminate approaches and mechanisms to study the mechanobiology of the cell surface in other cell types, including animal cells.

Plasticity, elasticity, and adhesion energy of plant cell walls: nanometrology of lignin loss using atomic force microscopy

The complex organic polymer, lignin, abundant in plants, prevents the efficient extraction of sugars from the cell walls that is required for large scale biofuel production. Because lignin removal is crucial in overcoming this challenge, the question of how the nanoscale properties of the plant cell ultrastructure correlate with delignification processes is important. Here, we report how distinct molecular domains can be identified and how physical quantities of adhesion energy, elasticity, and plasticity undergo changes, and whether such quantitative observations can be used to characterize delignification. By chemically processing biomass, and employing nanometrology, the various stages of lignin removal are shown to be distinguished through the observed morphochemical and nanomechanical variations. Such spatially resolved correlations between chemistry and nanomechanics during deconstruction not only provide a better understanding of the cell wall architecture but also is vital for devising optimum chemical treatments.

Formation of the Stomatal Outer Cuticular Ledge Requires a Guard Cell Wall Proline-Rich Protein.

Stomata are formed by a pair of guard cells which have thickened, elastic cell walls to withstand the large increases in turgor pressure that have to be generated to open the pore that they surround. We have characterized FOCL1, a guard cell-expressed, secreted protein with homology to Hyp-rich cell wall proteins. FOCL1-GFP localizes to the guard cell outer cuticular ledge and plants lacking FOCL1 produce stomata without a cuticular ledge. Instead the majority of stomatal pores are entirely covered over by a continuous fusion of the cuticle, and consequently plants have decreased levels of transpiration and display drought tolerance. The focl1 guard cells are larger and less able to reduce the aperture of their stomatal pore in response to closure signals suggesting that the flexibility of guard cell walls is impaired. FOCL1 is also expressed in lateral root initials where it aids lateral root emergence. We propose that FOCL1 acts in these highly specialized cells of the stomata and root to impart cell wall strength at high turgor and/or to facilitate interactions between the cell wall and the cuticle.

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity.

Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in β-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for β-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of β-(1,3)-glucan to the immune system occurs when the mannan layer is altered or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system.

Analyzing Cell Wall Elasticity After Hormone Treatment: An Example Using Tobacco BY-2 Cells and Auxin.

Atomic force microscopy, and related nano-indentation techniques, is a valuable tool for analyzing the elastic properties of plant cell walls as they relate to changes in cell wall chemistry, changes in development, and response to hormones. Within this chapter I will describe a method for analyzing the effect of the phytohormone auxin on the cell wall elasticity of tobacco BY-2 cells. This general method may be easily altered for different experimental systems and hormones of interest.

Sorption/desorption hysteresis revisited. Sorption properties of Pinus pinea L. analysed by the parallel exponential kinetics and Kelvin-Voigt models

Abstract The hygroscopic properties of Pinus pinea L. wood at 35 and 50°C were investigated by the dynamic vapour sorption (DVS) technique. The sorption kinetic behaviour was studied through the parallel exponential kinetics (PEK) model, which is subdivided into a fast and a slow process. The parameters obtained by PEK were interpreted based on the Kelvin-Voigt (KV) model to determine elasticity and viscosity values of the wood cell wall. The PEK data perfectly fit the experimental data. The temperature-dependent transition between the fast and slow processes is fluent. The slow process contributes more to the total hysteresis of sorption. The kinetic properties varied in relation to the type of cycle and the temperature. The moduli of elasticity and viscosity were higher in the slow process than in the fast one. In both processes, the moduli showed a decreasing tendency in relation to relative humidity.

Atomic Force Microscopy Studies of the Interaction of Antimicrobial Peptides with Bacterial Cells

Antimicrobial peptides (AMPs) are promising therapeutic alternatives to conventional antibiotics. Many AMPs are membrane-active but their mode of action in killing bacteria or in inhibiting their growth remains elusive. Recent studies indicate the mechanism of action depends on peptide structure and lipid components of the bacterial cell membrane. Owing to the complexity of working with living cells, most of these studies have been conducted with synthetic membrane systems, which neglect the possible role of bacterial surface structures in these interactions. In recent years, atomic force microscopy has been utilized to study a diverse range of biological systems under non-destructive, physiologically relevant conditions that yield in situ biophysical measurements of living cells. This approach has been applied to the study of AMP interaction with bacterial cells, generating data that describe how the peptides modulate various biophysical behaviours of individual bacteria, including the turgor pressure, cell wall elasticity, bacterial capsule thickness, and organization of bacterial adhesins.

A Mechanical Model to Interpret Cell-Scale Indentation Experiments on Plant Tissues in Terms of Cell Wall Elasticity and Turgor Pressure.

Morphogenesis in plants is directly linked to the mechanical elements of growing tissues, namely cell wall and inner cell pressure. Studies of these structural elements are now often performed using indentation methods such as atomic force microscopy. In these methods, a probe applies a force to the tissue surface at a subcellular scale and its displacement is monitored, yielding force-displacement curves that reflect tissue mechanics. However, the interpretation of these curves is challenging as they may depend not only on the cell probed, but also on neighboring cells, or even on the whole tissue. Here, we build a realistic three-dimensional model of the indentation of a flower bud using SOFA (Simulation Open Framework Architecture), in order to provide a framework for the analysis of force-displacement curves obtained experimentally. We find that the shape of indentation curves mostly depends on the ratio between cell pressure and wall modulus. Hysteresis in force-displacement curves can be accounted for by a viscoelastic behavior of the cell wall. We consider differences in elastic modulus between cell layers and we show that, according to the location of indentation and to the size of the probe, force-displacement curves are sensitive with different weights to the mechanical components of the two most external cell layers. Our results confirm most of the interpretations of previous experiments and provide a guide to future experimental work.

Dynamics of cell wall elasticity pattern shapes the cell during yeast mating morphogenesis.

The cell wall defines cell shape and maintains integrity of fungi and plants. When exposed to mating pheromone, Saccharomyces cerevisiae grows a mating projection and alters in morphology from spherical to shmoo form. Although structural and compositional alterations of the cell wall accompany shape transitions, their impact on cell wall elasticity is unknown. In a combined theoretical and experimental approach using finite-element modelling and atomic force microscopy (AFM), we investigated the influence of spatially and temporally varying material properties on mating morphogenesis. Time-resolved elasticity maps of shmooing yeast acquired with AFM in vivo revealed distinct patterns, with soft material at the emerging mating projection and stiff material at the tip. The observed cell wall softening in the protrusion region is necessary for the formation of the characteristic shmoo shape, and results in wider and longer mating projections. The approach is generally applicable to tip-growing fungi and plants cells.

Plumbing the depths: extracellular water storage in specialized leaf structures and its functional expression in a three-domain pressure -volume relationship.

A three-domain pressure-volume relationship (PV curve) was studied in relation to leaf anatomical structure during dehydration in the grey mangrove, Avicennia marina. In domain 1, relative water content (RWC) declined 13% with 0.85 MPa decrease in leaf water potential, reflecting a decrease in extracellular water stored primarily in trichomes and petiolar cisternae. In domain 2, RWC decreased by another 12% with a further reduction in leaf water potential to -5.1 MPa, the turgor loss point. Given the osmotic potential at full turgor (-4.2 MPa) and the effective modulus of elasticity (~40 MPa), domain 2 emphasized the role of cell wall elasticity in conserving cellular hydration during leaf water loss. Domain 3 was dominated by osmotic effects and characterized by plasmolysis in most tissues and cell types without cell wall collapse. Extracellular and cellular water storage could support an evaporation rate of 1 mmol m-2 s-1 for up to 54 and 50 min, respectively, before turgor loss was reached. This study emphasized the importance of leaf anatomy for the interpretation of PV curves, and identified extracellular water storage sites that enable transient water use without substantive turgor loss when other factors, such as high soil salinity, constrain rates of water transport.

Rootstock-induced physiological and biochemical mechanisms of drought tolerance in sweet orange

The poorly understood physiological and biochemical drought responses induced in sweet orange by citrus rootstocks of contrasting drought tolerance were investigated during a drought/rewatering cycle under controlled conditions. Long-term exposure of the grafted trees to a gradually increasing water deficit and subsequent recovery revealed distinct strategies of drought acclimation that were induced by the different rootstocks. Trees grafted onto the drought-tolerant rootstock ‘Cravo’ rangpur lime were less water conservative, exhibiting an increased cell-wall elasticity that contributes to turgor maintenance and its related processes of growth and photosynthesis over a wider range of soil–water potentials. On the other hand, the drought-tolerant ‘Sunki Tropical’ mandarin and drought-sensitive ‘Flying Dragon’ trifoliate orange rootstocks induced a water conservation strategy by increasing tissue rigidity under drought. ‘Sunki Tropical’ was also able to induce osmotic adjustment, conferring thereby a more efficient water conservation strategy than ‘Flying Dragon’ by allowing for turgor maintenance at lower soil–water potentials while attenuating cell dehydration and shrinkage. In contrast to ‘Cravo’ and ‘Sunki Tropical’, trees grafted onto ‘Flying Dragon’ exhibited a significant photoinhibition of the photosystem II reaction centers, as well as an increased H2O2 production and lipid peroxidation under drought treatment. A significantly higher activity of the antioxidant enzyme GPX was also observed in drought stressed trees grafted onto ‘Flying Dragon’. Collectively, these results support the involvement of elastic and osmotic adjustments, as well as the control of oxidative stress, as functional leaf traits associated with the rootstock-induced drought tolerance in sweet orange.

Freezing resistance in Patagonian woody shrubs: the role of cell wall elasticity and stem vessel size.

Freezing resistance through avoidance or tolerance of extracellular ice nucleation is important for plant survival in habitats with frequent subzero temperatures. However, the role of cell walls in leaf freezing resistance and the coordination between leaf and stem physiological processes under subzero temperatures are not well understood. We studied leaf and stem responses to freezing temperatures, leaf and stem supercooling, leaf bulk elastic modulus and stem xylem vessel size of six Patagonian shrub species from two sites (plateau and low elevation sites) with different elevation and minimum temperatures. Ice seeding was initiated in the stem and quickly spread to leaves, but two species from the plateau site had barriers against rapid spread of ice. Shrubs with xylem vessels smaller in diameter had greater stem supercooling capacity, i.e., ice nucleated at lower subzero temperatures. Only one species with the lowest ice nucleation temperature among all species studied exhibited freezing avoidance by substantial supercooling, while the rest were able to tolerate extracellular freezing from -11.3 to -20 °C. Leaves of species with more rigid cell walls (higher bulk elastic modulus) could survive freezing to lower subzero temperatures, suggesting that rigid cell walls potentially reduce the degree of physical injury to cell membranes during the extracellular freezing and/or thaw processes. In conclusion, our results reveal the temporal-spatial ice spreading pattern (from stem to leaves) in Patagonian shrubs, and indicate the role of xylem vessel size in determining supercooling capacity and the role of cell wall elasticity in determining leaf tolerance of extracellular ice formation.

Contributions of Basic Chemical Components to the Mechanical Behavior of Wood Fiber Cell Walls as Evaluated by Nanoindentation

Selective chemical extraction was applied to gradually remove classes of chemical components from wood cell walls. Nanoindentation was performed on the control and treated wood cell walls to evaluate the contributions of the chemical components to the cell walls by measuring the elastic modulus, hardness, and creep compliance. Burger’s model was applied to simulate the process of nanoindentation and to gain insight into the response of visco-elastic properties to the chemical components. Wood extractives showed limited effects on the cell-wall mechanics; however, the removal of hemicelluloses and lignin resulted in reductions of 11.7% and 28.4%, respectively, in the elastic modulus and 14.8% and 30.4%, respectively, in the hardness. The extraction of hemicelluloses and lignin reduced the resistance of wood cell walls to creep. Furthermore, the extracted parameters from Burger’s modeling indicated that cellulose exhibited the greatest influence on the mechanical properties of wood cell wall, while hemicelluloses exhibited the greatest contribution to cell-wall viscosity, and lignin contributed extensively to cell-wall elasticity.

On the Influence of the Local Cell Wall Elasticity on the Cell Shape during Yeast Mating Morphogenesis

The cell wall of Saccharomyces cerevisiae is the first barrier of cell against the environment and furthermore defines the cell shape by maintaining the structural integrity. During cell proliferation the shape of the cell is spherical to ellipsoid, but changes decisively as a response to a mating pheromone. Here the pheromone induces a directed growth of the cell towards the highest pheromone gradient, resulting in an elongated cell shape, called shmoo. The transition of the cell shape is associated with alterations of the structure and composition of the cell wall, however the underlying mechanism remains unclear.To elucidate the influence of the material properties of the cell wall, especially the elasticity, on the shape evolution of shmooing yeast, we applied a combined theoretical and experimental approach, using finite element modeling and atomic force microscopy. Nano-indentation experiments on the cell wall of living and shmooing yeasts cells revealed a characteristic and time resolved elasticity pattern. Inspired by those insights, we built a dynamic cell wall model, which was able to reproduce the formation of the characteristic shmoo shape and helped us to understand the necessity of locally varying cell wall elasticity.

Wall mechanics and exocytosis define the shape of growth domains in fission yeast.

The amazing structural variety of cells is matched only by their functional diversity, and reflects the complex interplay between biochemical and mechanical regulation. How both regulatory layers generate specifically shaped cellular domains is not fully understood. Here, we report how cell growth domains are shaped in fission yeast. Based on quantitative analysis of cell wall expansion and elasticity, we develop a model for how mechanics and cell wall assembly interact and use it to look for factors underpinning growth domain morphogenesis. Surprisingly, we find that neither the global cell shape regulators Cdc42-Scd1-Scd2 nor the major cell wall synthesis regulators Bgs1-Bgs4-Rgf1 are reliable predictors of growth domain geometry. Instead, their geometry can be defined by cell wall mechanics and the cortical localization pattern of the exocytic factors Sec6-Syb1-Exo70. Forceful re-directioning of exocytic vesicle fusion to broader cortical areas induces proportional shape changes to growth domains, demonstrating that both features are causally linked.

Cell Wall Remodeling Enzymes Modulate Fungal Cell Wall Elasticity and Osmotic Stress Resistance.

ABSTRACTThe fungal cell wall confers cell morphology and protection against environmental insults. For fungal pathogens, the cell wall is a key immunological modulator and an ideal therapeutic target. Yeast cell walls possess an inner matrix of interlinked β-glucan and chitin that is thought to provide tensile strength and rigidity. Yeast cells remodel their walls over time in response to environmental change, a process controlled by evolutionarily conserved stress (Hog1) and cell integrity (Mkc1, Cek1) signaling pathways. These mitogen-activated protein kinase (MAPK) pathways modulate cell wall gene expression, leading to the construction of a new, modified cell wall. We show that the cell wall is not rigid but elastic, displaying rapid structural realignments that impact survival following osmotic shock. Lactate-grown Candida albicans cells are more resistant to hyperosmotic shock than glucose-grown cells. We show that this elevated resistance is not dependent on Hog1 or Mkc1 signaling and that most cell death occurs within 10 min of osmotic shock. Sudden decreases in cell volume drive rapid increases in cell wall thickness. The elevated stress resistance of lactate-grown cells correlates with reduced cell wall elasticity, reflected in slower changes in cell volume following hyperosmotic shock. The cell wall elasticity of lactate-grown cells is increased by a triple mutation that inactivates the Crh family of cell wall cross-linking enzymes, leading to increased sensitivity to hyperosmotic shock. Overexpressing Crh family members in glucose-grown cells reduces cell wall elasticity, providing partial protection against hyperosmotic shock. These changes correlate with structural realignment of the cell wall and with the ability of cells to withstand osmotic shock.

Morphogenesis of the Fission Yeast Cell through Cell Wall Expansion

The shape of walled cells such as fungi, bacteria, and plants are determined by the cell wall. Models for cell morphogenesis postulate that the effects of turgor pressure and mechanical properties of the cell wall can explain the shapes of these diverse cell types. However, in general, these models await validation through quantitative experiments. Fission yeast Schizosaccharomyces pombe are rod-shaped cells that grow by tip extension and then divide medially through formation of a cell wall septum. Upon cell separation after cytokinesis, the new cell ends adopt a rounded morphology. Here, we show that this shape is generated by a very simple mechanical-based mechanism in which turgor pressure inflates the elastic cell wall in the absence of cell growth. This process is independent of actin and new cell wall synthesis. To model this morphological change, we first estimate the mechanical properties of the cell wall using several approaches. The lateral cell wall behaves as an isotropic elastic material with a Young's modulus of 50 ± 10 MPa inflated by a turgor pressure estimated to be 1.5 ± 0.2 MPa. Based upon these parameters, we develop a quantitative mechanical-based model for new end formation that reveals that the cell wall at the new end expands into its characteristic rounded shape in part because it is softer than the mature lateral wall. These studies provide a simple example of how turgor pressure expands the elastic cell wall to generate a particular cell shape.