Wall Polymers Research Articles

Vacuum-heat treatment of Scots pine (Pinus sylvestris L.) wood pretreated with propanetriol

ABSTRACT Scots pine sapwood was pretreated with two levels of propanetriol (20% and 40% w/w glycerol), and then subjected to vacuum-heat treatment (VHT) at 180°C and 200°C. The treated samples were examined with respect to their weight and volumetric changes, mechanical properties, colour changes, and dynamic water vapour sorption. The weight of the samples after VHT did not change with increasing the temperature, but it was increased in glycerol pretreated samples. Combination of glycerol pretreatment and VHT decreased the maximum swelling. Total colour change was significantly higher during VHT at a higher temperature, while no obvious trend observed in the samples pretreated with glycerol. Modulus of elasticity (MOE) and modulus of rupture (MOR) were not affected by solely VHT, but strongly decreased after glycerol pretreatment. The equilibrium moisture content (EMC) of the samples decreased by VHT. The glycerol pretreatment caused a reduction in EMC values at a relative humidity (RH) below 60%, but considerably increased the moisture sorption in the RH above 75%. VHT slightly reduced the sorption hysteresis compared to untreated wood, but an apparent reduction in hysteresis observed by glycerol pretreatment. This indicates that the flexibility of the wood cell wall polymers increases due to glycerol pretreatment, which results in decreased MOE and sorption hysteresis values.

Polymerization in Shear Flow: From Bowl-Shaped Glyco-Microcarriers to Self-Propelled Micromotors.

We report a simple and effective method to synthesize composite bowl-shaped colloidal particles composed of a magnetic bottom and polymer wall. The bowl-shape is determined by the stirring and the resulting centrifugal force acting on the components during the synthesis. By systematically varying the stirring speed, we have found the lower and upper bounds for the bowl formation. Moreover, the size and thickness of the bowl can be tuned by changing the amount of the TPM monomer during synthesis. Functional groups can be easily introduced in the solidifying step through the polymerization of TPM by adding initiators and functional monomers. For example, by copolymerization of glycomonomer with TPM, bowl-shaped particles with biological activity can be obtained. In addition, such composite particles can be further modified to produce micromotors with well-controlled motions.

Seed hemicelluloses tailor mucilage properties and salt tolerance.

While Arabidopsis seed coat epidermal cells have become an excellent genetic system to study the biosynthesis and structural roles of various cell wall polymers, the physiological function of the secreted mucilaginous polysaccharides remains ambiguous. Seed mucilage is shaped by two distinct classes of highly substituted hemicelluloses along with cellulose and structural proteins, but their interplay has not been explored. We deciphered the functions of four distinct classes of cell wall polymers by generating a series of double mutants with defects in heteromannan, xylan, cellulose, or the arabinogalactan protein SALT-OVERLY SENSITIVE 5 (SOS5), and evaluating their impact on mucilage architecture and seed germination during salt stress. We discovered that muci10 seeds, lacking heteromannan branches, had elevated tolerance to salt stress, while heteromannan elongation mutants exhibited reduced germination in calcium chloride (CaCl2 ). By contrast, xylan made by MUCILAGE-RELATED21 (MUCI21) was found to be required for the adherence of mucilage pectin to microfibrils made by CELLULOSE SYNTHASE5 (CESA5) as well as to a SOS5-mediated network. Our results indicate that the substitution of xylan and glucomannan in seeds can fine-tune mucilage adherence and salt tolerance, respectively. The study of germinating seeds can thus provide insights into the synthesis, modification and function of complex glycans.

Viscoelastic properties of thermo-hydro-mechanically treated beech (Fagus sylvatica L.) determined using dynamic mechanical analysis

Thermo-hydro-mechanical (THM) processing can improve the intrinsic properties of wood, produce new materials, and give desired form and function to new applications. THM treatments change the mechanical properties of wood and may change its viscoelastic properties as well. Therefore, the objective of this study was to assess the viscoelastic properties of THM-treated wood at several humidity and load levels. To explore these changes, this study applied a THM treatment to beech (Fagus sylvatica L.) wood with steam (620 kPa) and heat (170 °C), followed by densification and increased temperature (200 °C) in a hot-press, which was then cooled while under pressure. Two initial specimen thicknesses before THM treatment were used to study the difference between density ratios. Specimens were tested in a humidity-controlled dynamic mechanical analyser (DMA) to apply creep stress with different loading levels (20% and 30% of expected modulus of rupture) and relative humidity levels (30%, 50%, and 65% RH). The creep compliance/recovery response was monitored, and dynamic moduli were measured before and after the application of creep stress. The loss modulus measured was highest for specimens tested at 65% RH and lowest in specimens tested at 30% RH, which is a direct result of the viscous response of the material. Increased damping of the specimens was also observed at higher RH, which is typical for wood products due to added moisture in the cell wall acting as a plasticizer to cell wall polymers. Like previous studies, THM treatment lowered the equilibrium moisture content (EMC) of densified wood specimens, which affected their mechanical performance. THM treatment yields higher strength and lower EMC, suggesting that this product could be suitable for structural applications where their service life is in an indoor environment. THM treatment also resulted in decreased creep compliance and recovery compliance as compared to control specimens.

Systems identification and characterization of \u03b2-glucuronosyltransferase genes involved in arabinogalactan-protein biosynthesis in plant genomes

Utilizing plant biomass for bioethanol production requires an understanding of the molecular mechanisms involved in plant cell wall assembly. Arabinogalactan-proteins (AGPs) are glycoproteins that interact with other cell wall polymers to influence plant growth and developmental processes. Glucuronic acid, which is transferred to the AGP glycan by β-glucuronosyltransferases (GLCATs), is the only acidic sugar in AGPs with the ability to bind calcium. We carried out a comprehensive genome-wide analysis of a putative GLCAT gene family involved in AGP biosynthesis by examining its sequence diversity, genetic architecture, phylogenetic and motif characteristics, selection pressure and gene expression in plants. We report the identification of 161 putative GLCAT genes distributed across 14 plant genomes and a widely conserved GLCAT catalytic domain. We discovered a phylogenetic clade shared between bryophytes and higher land plants of monocot grass and dicot lineages and identified positively selected sites that do not result in functional divergence of GLCATs. RNA-seq and microarray data analyses of the putative GLCAT genes revealed gene expression signatures that likely influence the assembly of plant cell wall polymers which is critical to the overall growth and development of edible and bioenergy crops.

Novel polysulfone ultrafiltration membranes incorporating polydopamine functionalized graphene oxide with enhanced flux and fouling resistance

Novel PSF composite UF membranes incorporating low loadings of polydopamine-functionalized graphene oxide particles (rGO-PDA) were fabricated and investigated. The functionalization was confirmed using FTIR-UATR, Raman spectra, XPS, and SEM. Pristine PSF, PSF/GO, and PSF/rGO-PDA MMMs were then prepared using the phase inversion technique and analysed using FTIR, SEM, AFM, and contact angle (CA). The cross-section SEM images showed better distribution of rGO-PDA particles in the pores and polymer wall whereas the pristine GO particles aggregate and partially block the pores. Thus, the pure water flux increased with the addition of rGO-PDA without affecting the rejection properties, while the flux decreased with the embedding of pristine GO particles. The highest pure water permeability (PWP) was obtained with PSF/rGO-PDA-0.1 to be approximately twice that of the pristine PSF and PSF/GO-0.1. All membranes exhibited complete rejection of BSA and HA, and showed almost similar performance against different dyes. The FRRs of the pristine PSF after three fouling cycles (FRR3) against BSA and HA were recorded to be 57.8% and 70.7% respectively. FRR3 was enhanced by around 30% with PSF/rGO-PDA composites. The MMMs prepared in this work are expected to have great potential on ultrafiltration and similar studies on other membrane processes.

A novel mechanism of β-glucosidase stimulation through a monosaccharide binding-induced conformational change

It is urgent the transition from a fossil fuel-based economy to a sustainable bioeconomy based on bioconversion technologies using renewable plant biomass feedstocks to produce high chemicals, bioplastics, and biofuels. β-Glucosidases are key enzymes responsible for degrading the plant cell wall polymers, as they cleave glucan-based oligo- and polysaccharides to generate glucose. Monosaccharide-tolerant or -stimulated β-glucosidases have been reported in the past decade. Here, we describe a novel mechanism of β-glucosidase stimulation by glucose and xylose. The glycoside hydrolase 1 family β-glucosidase from Thermotoga petrophila (TpBgl1) displays a typical glucose stimulation mechanism based on an increased Vmax and decreased Km in response to glucose. Through molecular docking and dynamics analyses, we mapped putative monosaccharide binding regions (BRs) on the surface of TpBgl1. Our results indicate that after interaction with glucose or xylose at BR1 site, an adjacent loop region assumes an extended conformation, which increases the entrance to the TpBgl1 active site, improving product formation. Biochemical assays with TpBgl1 BR1 mutants, TpBgl1D49A/Y410A and TpBgl1D49K/Y410H, resulted in decreasing and abolishing monosaccharide stimulation, respectively. These mutations also impaired the BR1 looping extension responsible for monosaccharide stimulation. This study provides a molecular basis for the rational design of β-glucosidases for biotechnological applications.

Cycling in degradation of organic polymers and uptake of nutrients by a litter-degrading fungus.

SummaryWood and litter degrading fungi are the main decomposers of lignocellulose and thus play a key role in carbon cycling in nature. Here, we provide evidence for a novel lignocellulose degradation strategy employed by the litter degrading fungus Agaricus bisporus (known as the white button mushroom). Fusion of hyphae allows this fungus to synchronize the activity of its mycelium over large distances (50 cm). The synchronized activity has a 13‐h interval that increases to 20 h before becoming irregular and it is associated with a 3.5‐fold increase in respiration, while compost temperature increases up to 2°C. Transcriptomic analysis of this burst‐like phenomenon supports a cyclic degradation of lignin, deconstruction of (hemi‐) cellulose and microbial cell wall polymers, and uptake of degradation products during vegetative growth of A. bisporus. Cycling in expression of the ligninolytic system, of enzymes involved in saccharification, and of proteins involved in nutrient uptake is proposed to provide an efficient way for degradation of substrates such as litter.

Differential localization of cell wall polymers across generations in the placenta of Marchantia polymorpha.

To further knowledge on cell wall composition in early land plants, we localized cell wall constituents in placental cells of the liverwortMarchantia polymorpha L. using monoclonal antibodies (MAbs) in the transmission electron microscope and histochemical staining. The placenta ofM. polymorphais similar to the majority of bryophytes in thatboth generationscontain transfer cells with extensive wall ingrowths.Although the four major cell wall polymers, i.e., cellulose, pectins, hemicelluloses, and arabinogalactan proteins, are present, there are variations in the richness and specificity across generations. An abundance of homogalacturonan pectins in all placental cell walls is consistent with maintaining cell wall permeability and an acidic apoplastic pH necessary for solute transport.Although similar in ultrastructure, transfer cell walls on the sporophyte side in M. polymorpha are enriched with xyloglucans and diverse AGPs not detected on the gametophyte side of the placenta. Gametophyte wall ingrowths are more uniform in polymer composition. Lastly, extensins and callose are not components of transfer cell walls of M. polymorpha,which deviates from studies on transfer cells in other plants. The difference in polymer localizations in transfer cell walls between generations is consistent with directional movement from gametophyte to sporophyte in this liverwort.

Targeted Metagenomics of Retting in Flax: The Beginning of the Quest to Harness the Secret Powers of the Microbiota.

The mechanical and chemical properties of natural plant fibers are determined by many different factors, both intrinsic and extrinsic to the plant, during growth but also after harvest. A better understanding of how all these factors exert their effect and how they interact is necessary to be able to optimize fiber quality for use in different industries. One important factor is the post-harvest process known as retting, representing the first step in the extraction of bast fibers from the stem of species such as flax and hemp. During this process microorganisms colonize the stem and produce hydrolytic enzymes that target cell wall polymers thereby facilitating the progressive destruction of the stem and fiber bundles. Recent advances in sequencing technology have allowed researchers to implement targeted metagenomics leading to a much better characterization of the microbial communities involved in retting, as well as an improved understanding of microbial dynamics. In this paper we review how our current knowledge of the microbiology of retting has been improved by targeted metagenomics and discuss how related ‘-omics’ approaches might be used to fully characterize the functional capability of the retting microbiome.

Molecular Logic of Prokaryotic Surface Layer Structures.

Most prokaryotic cells are encased in a surface layer (S-layer) consisting of a paracrystalline array of repeating lattice-forming proteins. S-layer proteins populate a vast and diverse sequence space, performing disparate functions in prokaryotic cells, including cellular defense, cell-shape maintenance, and regulation of import and export of materials. This article highlights recent advances in the understanding of S-layer structure and assembly, made possible by rapidly evolving structural and cell biology methods. We underscore shared assembly principles revealed by recent work and discuss a common molecular framework that may be used to understand the structural organization of S-layer proteins across bacteria and archaea.

Comparative Analysis of the Lignification Process of Two Bamboo Shoots Stored at Room Temperature.

Two types of bamboo shoots, high bamboo (Phyllostachys prominens) shoots (HBSes) and moso bamboo (Phyllostachys edulis) shoots (MBSes), underwent a fast post-harvest lignification process under room temperature storage. To explore the mechanism of lignification in two types of bamboo shoots after post-harvest during room temperature storage, the measurement of cell wall polymers (lignin and cellulose) and enzyme activities of phenylalanine ammonialyase (PAL) and peroxidase (POD), and relative expression of related transcription networks factors (TFs) were performed. The results suggested that the lignification process in HBSes is faster than that in MBSes because of incremental increase in lignin and cellulose contents within 6 days and the shorter shelf-life. Additionally, compared with the expression pattern of lignification-related TFs and correlation analysis of lignin and cellulose contents, MYB20, MYB43, MYB85 could function positively in the lignification process of two types of bamboo shoots. A negative regulator, KNAT7, could negatively regulate the lignin biosynthesis in two types of bamboo shoots. In addition, MYB63 could function positively in HBSes, and NST1 could function negatively in MBSes. Notably, MYB42 may function differently in the two types of bamboo shoots, that is, a positive regulator in HBSes, but a negative regulator in MBSes. Transcription networks provide a comprehensive analysis to explore the mechanism of lignification in two types of bamboo shoots after post-harvest during room temperature storage. These results suggest that the lignification of bamboo shoots was mainly due to the increased activity of POD, higher expression levels of MYB20, MYB43, MYB63, and MYB85 genes, and lower expression levels of KNAT7 and NST1 genes, and the lignification process of HBSes and MBSes had significant differences.

Fungal Pretreatments on Non-Sterile Solid Digestate to Enhance Methane Yield and the Sustainability of Anaerobic Digestion

Fungi can run feedstock pretreatment to improve the hydrolysis and utilization of recalcitrant lignocellulose-rich biomass during anaerobic digestion (AD). In this study, three fungal strains (Coprinopsis cinerea MUT 6385, Cyclocybe aegerita MUT 5639, Cephalotrichum stemonitis MUT 6326) were inoculated in the non-sterile solid fraction of digestate, with the aim to further (re)use it as a feedstock for AD. The application of fungal pretreatments induced changes in the plant cell wall polymers, and different profiles were observed among strains. Significant increases (p < 0.05) in the cumulative biogas and methane yields with respect to the untreated control were observed. The most effective pretreatment was carried out for 20 days with C. stemonitis, causing the highest hemicellulose, lignin, and cellulose reduction (59.3%, 9.6%, and 8.2%, respectively); the cumulative biogas and methane production showed a 182% and 214% increase, respectively, compared to the untreated control. The increase in AD yields was ascribable both to the addition of fungal biomass, which acted as an organic feedstock, and to the lignocellulose transformation due to fungal activity during pretreatments. The developed technologies have the potential to enhance the anaerobic degradability of solid digestate and untap its biogas potential for a further digestion step, thus allowing an improvement in the environmental and economic sustainability of the AD process and the better management of its by-products.

Probing the role of cell wall feruloylation during maize development by differential expression of an apoplast targeted fungal ferulic acid esterase.

While many aspects of the growth of maize are well understood, the role of cell wall feruloylation particularly during internode elongation has not been firmly established, but results so far indicate that it has significant implications for both biofuel feedstock conversion and for crop yield. The growth of the cell wall is achieved by synthesis, integration and cross-linking between wall polymers. As ferulate oxidative coupling of arabinoxylan side chains constitutes a significant type of cross-link in grass cell walls, it is expected to have a crucial role in plant growth. Making use of plants expressing an apoplast targeted Aspergillus niger FAEA under the control of either a constitutive or an inducible promoter, the role of cell wall feruloylation in maize internode expansion was investigated. Analysis of FAEA expressing plants showed that where FAEA was targeted to the apoplast under a constitutive promoter, plants varied in stature either from semi-dwarf plants with a 40–60% height reduction, to extreme dwarf mutants with over 90% reduction in plant heights compared to controls. Results indicate that disruption of cell wall feruloylation by FAEA occurs before the start of rapid internode expansion is initiated and affects the normal course of internode elongation, resulting in short internodes and dwarfed plants. In contrast, when under the inducible Lm See1 senescence promoter, FAEA activity was found to be low up to the VT stage of development but increased significantly at the VR stage as plants began to senesce, strongly suggesting that normal cell wall feruloylation is required for the process of internode expansion. In addition, with apoplast targeted expression of FAEA under control of the senescence enhanced promoter it was possible to demonstrate decreased cell wall feruloylation without affecting internode expansion or other aspects of plant development.

Forward Flux Sampling of Polymer Desorption Paths from a Solid Surface into Dilute Solution.

We compute desorption rates for isolated polymers adsorbed to a solid wall with a rare event sampling technique called multilevel splitting, also known as forward flux sampling. We interpret computed rates with theories based on the conjecture that the product of the desorption time and diffusivity divided by squared radius of gyration scales with exp(h/Rg) where h is the equilibrium ratio of adsorbed surface concentration of polymer to bulk concentration of polymer . As the polymer–wall interaction energy is increased, the slope of vs. nearly approaches unity, as expected for strongly-adsorbing chains, where N is the degree of polymerization and is the height-averaged monomer–wall interaction energy for a strongly adsorbed chain. However, we also find that this scaling law is only accurate when adsorption strength per monomer exceeds a threshold value on the order of 0.3–0.5 kBT for a freely jointed chain without or with excluded volume effects. Below the critical value, we observe that becomes nearly constant with N, so that , with . This suggests a crossover from “strong” detachment-controlled to a “weak” diffusion-controlled desorption rate as VMF/kBT drops below some threshold. These results may partially explain experimental data, that in some cases show “strong” exponential dependence of desorption time on chain length, while in others a “weak” power-law dependence is found. However, in the “strong” adsorption case, our results suggest much longer desorption times than those measured, while the reverse is true in the weak adsorption limit. We discuss possible reasons for these discrepancies.

Single-Cell Elasticity Measurement with an Optically Actuated Microrobot.

A cell elasticity measurement method is introduced that uses polymer microtools actuated by holographic optical tweezers. The microtools were prepared with two-photon polymerization. Their shape enables the approach of the cells in any lateral direction. In the presented case, endothelial cells grown on vertical polymer walls were probed by the tools in a lateral direction. The use of specially shaped microtools prevents the target cells from photodamage that may arise during optical trapping. The position of the tools was recorded simply with video microscopy and analyzed with image processing methods. We critically compare the resulting Young’s modulus values to those in the literature obtained by other methods. The application of optical tweezers extends the force range available for cell indentations measurements down to the fN regime. Our approach demonstrates a feasible alternative to the usual vertical indentation experiments.

Wood forming tissue-specific expression of PdSuSy and HCHL increases holocellulose content and improves saccharification in Populus

Development of strategies to deconstruct lignocellulosic biomass in tree species is essential for biofuels and biomaterials production. We applied a wood forming tissue-specific system in a hybrid poplar to express both PdSuSy (a sucrose synthase gene from Populus deltoides × P. euramericana that has not been functionally characterized) and HCHL (the hydroxycinnamoyl-CoA hydratase-lyase gene from Pseudomonas fluorescens, which inhibits lignin polymerization in Arabidopsis). The PdSuSy-HCHL overexpression poplars correspondingly driven by the promoters of Arabidopsis AtCesA7 and AtC4H resulted in a significant increase in cellulose (> 8%), xylan (> 12%) and glucose (> 29%) content, accompanying a reduction in galacturonic acid (> 36%) content, compared to control plants. The saccharification efficiency of these overexpression poplars was dramatically increased by up to 27%, but total lignin content was unaffected. These transgenic poplars showed inhibited growth characteristics, including > 16% reduced plant height, > 10% reduced number of internodes, and > 18% reduced fresh weight after growth of 4 months, possibly due to relatively low expression of HCHL in secondary xylem. Our results demonstrate the structural complexity and interaction of the cell wall polymers in wood tissue and outline a potential method to increase biomass saccharification in woody species.

Sorption behavior and hydroxyl accessibility of wood treated with different cyclic N-methylol compounds

Cyclic N-methylol compounds such as 1,3-dimethylol-4,5-dihydroxyethyleneurea (DMDHEU) have been used to modify wood and prevent negative effects related to the uptake of moisture. However, the changes in the sorption behavior of wood by treatments with DMDHEU and its derivatives are not fully understood. In the present study, wood blocks were treated with DMDHEU, ether-modified DMDHEU and diethyleneglycolated DMDHEU in order to study the factors that control the changes in moisture uptake in the hygroscopic range (0–95% RH). Dimensional changes of wood blocks during water soaking cycles suggested that the treatments caused a permanent cell wall bulking, whereas the swelling restraint by cross-linking of adjacent cell wall polymers was not permanent. However, the changes in water vapor sorption were not only a result of the cell wall bulking effect that reduced the space in the cell wall to accommodate water. The N-methylol compounds within the wood also provided additional sorption sites, but there was no correlation between absorbed water and accessible OH groups. It was speculated that the co-condensation of the N-methylol compounds with wood polymers had a significant effect on the sorption of the treated wood. At elevated RH, pure resins that were formed by self-condensation took up large quantities of moisture. However, when the N-methylol compounds were heat-cured within the hierarchical structure of wood, the moisture uptake of the treated wood at elevated RH was even lower compared to unmodified wood. Furthermore, the covalent bond formation between wood and resin prolonged the attainment of an equilibrium moisture content.

Mild Acetylation and Solubilization of Ground Whole Plant Cell Walls in EmimAc: A Method for Solution-State NMR in DMSO-d6.

Lignocellulosic biomass is mainly composed of polysaccharides and lignin. The complexity and diversity of the plant cell wall polymers makes it difficult to isolate the components in pure form for characterization. Many current approaches to analyzing the structure of lignocellulose, which involve sequential extraction and characterization of the resulting fractions, are time-consuming and labor-intensive. The present study describes a new and facile system for rationally derivatizing and dissolving coarsely ground plant cell wall materials. Using ionic liquids (EmimAc) and dichloroacetyl chloride as a solvent/reagent produced mildly acetylated whole cell walls without significant degradation. The acetylated products were soluble in DMSO-d6 from which they can be characterized by solution-state two-dimensional nuclear magnetic resonance (2D NMR) spectrometry. A distinct advantage of the procedure is that it realizes the dissolution of whole lignocellulosic materials without requiring harsh ball milling, thereby allowing the acquisition of high-resolution 2D NMR spectra to revealing structural details of the main components (lignin and polysaccharides). The method is therefore beneficial to understanding the composition and structure of biomass aimed at its improved utilization.

Mutations in the Pectin Methyltransferase QUASIMODO2 Influence Cellulose Biosynthesis and Wall Integrity in Arabidopsis.

Pectins are abundant in the cell walls of dicotyledonous plants, but how they interact with other wall polymers and influence wall integrity and cell growth has remained mysterious. Here, we verified that QUASIMODO2 (QUA2) is a pectin methyltransferase and determined that QUA2 is required for normal pectin biosynthesis. To gain further insight into how pectin affects wall assembly and integrity maintenance, we investigated cellulose biosynthesis, cellulose organization, cortical microtubules, and wall integrity signaling in two mutant alleles of Arabidopsis (Arabidopsis thaliana) QUA2, qua2 and tsd2 In both mutants, crystalline cellulose content is reduced, cellulose synthase particles move more slowly, and cellulose organization is aberrant. NMR analysis shows higher mobility of cellulose and matrix polysaccharides in the mutants. Microtubules in mutant hypocotyls have aberrant organization and depolymerize more readily upon treatment with oryzalin or external force. The expression of genes related to wall integrity, wall biosynthesis, and microtubule stability is dysregulated in both mutants. These data provide insights into how homogalacturonan is methylesterified upon its synthesis, the mechanisms by which pectin functionally interacts with cellulose, and how these interactions are translated into intracellular regulation to maintain the structural integrity of the cell wall during plant growth and development.