Cell Wall Hemicellulose Research Articles

Xyloglucan Deficiency Disrupts Microtubule Stability and Cellulose Biosynthesis in Arabidopsis, Altering Cell Growth and Morphogenesis.

Xyloglucan constitutes most of the hemicellulose in eudicot primary cell walls and functions in cell wall structure and mechanics. Although Arabidopsis (Arabidopsis thaliana) xxt1 xxt2 mutants lacking detectable xyloglucan are viable, they display growth defects that are suggestive of alterations in wall integrity. To probe the mechanisms underlying these defects, we analyzed cellulose arrangement, microtubule patterning and dynamics, microtubule- and wall-integrity-related gene expression, and cellulose biosynthesis in xxt1 xxt2 plants. We found that cellulose is highly aligned in xxt1 xxt2 cell walls, that its three-dimensional distribution is altered, and that microtubule patterning and stability are aberrant in etiolated xxt1 xxt2 hypocotyls. We also found that the expression levels of microtubule-associated genes, such as MAP70-5 and CLASP, and receptor genes, such as HERK1 and WAK1, were changed in xxt1 xxt2 plants and that cellulose synthase motility is reduced in xxt1 xxt2 cells, corresponding with a reduction in cellulose content. Our results indicate that loss of xyloglucan affects both the stability of the microtubule cytoskeleton and the production and patterning of cellulose in primary cell walls. These findings establish, to our knowledge, new links between wall integrity, cytoskeletal dynamics, and wall synthesis in the regulation of plant morphogenesis.

English

This study characterizes the cell wall hemicellulose and pectins polymers of Retama raetam. This species develops a particularly important root system and is adapted to arid areas. The cellulose, hemicelluloses and pectins were extracted. The cellulose remains the major component of the wall (27% for young roots and 80% for  adult roots), hemicelluloses (14.3% for young roots and 3.6%  for adult roots) and pectins (17.3% for young roots and 4.1% for adult roots). The monosaccharidic composition of water soluble extracts determined by gas liquid chromatography (GLC) and completed by infrared (FTIR) spectroscopy of hemicellulosic shows the presence of xylose as a major monosaccharide in the non-cellulose polysaccharides (47.8% for young roots and 59.5% for adult roots). These results indicate the presence of the homogalacturonans and rhamnogalacturonans in pectin. This study constitutes the preliminary data obtained in the biochemical analysis of the parietal compounds of the roots of a species which grows in an arid area in comparison with those of its aerial parts.   Key words: Retama  raetam,  roots, cell wall, investigation,  polysaccharides, monosaccharidic.

Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose.

Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a β-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded ~70% more ethanol compared with wild type. Plants expressing 35S:AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S:AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.

Rate of Decomposition of Plant Litter and Factors Affecting it

The plant materials largely consist of lignified cell wall materials viz cellulose, hemi-cellulose and lignin being the three major structural components. The present study revealed the lignocellulosic factors affecting the decomposition of different plant residues. The litters of different plants do not decompose at the same rate under similar environmental conditions. Coniferous litter decomposes very slowly upto 31 years as compared to deciduous litters that decompose rapidly, considerable difference exists between different types. The difference may be due to difference in anatomy of litters, nutrient status and the possible production of substances toxic to microorganisms. Even the two different parts of the same plant, may have different rates of decomposition. The nutrient contents of plant litters have significant correlation with the rate of decomposition. Addition of nitrogen, phosphorus and iron accelerated the rate of decomposition in optimum concentration. The rate of decomposition is also affected with placement of litter as incorporated litter in to soil decompose more rapidly as compared to those left on the soil surface. The ratio of pentosans to lignin also controls the rate of decomposition of litters; if the ratio of pentosans to lignin was more than 1, the rate of decomposition will be faster; if 1.0–0.5 then the rate of decomposition will be slow and if 0.5–0.3 then, it will be decomposed very slowly. The optimum temperature and the moisture have been found to be a major controlling factor in rate of litter decomposition.

Xylan Degrading Enzymes from Fungal Sources

Fungi have the ability to degrade xylan as the major component of plant cell wall hemicellulose. Fungi have evolved batteries of xylanolytic enzymes that concertedly act to depolymerise xylan backbones decorated with variable carbohydrate branches. As an alternative to acid extraction in industrial processes the combination of endo-1,4-β-xylanase and β-xylosidase can reduce xylan to xylose. However, unlike chemical extraction procedures enzyme systems can selectively hydrolyse α-L-arabinofuranosyl, 4-O-methyl-α-D-glucuronopyranosyl, acetyl and phenolic branches, and therefore have the potential to deconstruct hemicellulose whilst retaining desirable structural integrity and functionality. The sources, structures and catalytic activities of fungal xylanolytic enzymes are reviewed and discussed in the context of their biotechnological potential.

Crystallization and preliminary X-ray crystallographic analysis of a putative acetylxylan esterase from Talaromyces cellulolyticus.

Acetylxylan esterase (AXE) catalyzes the hydrolytic cleavage of the ester bond between acetic acid and hemicellulose in plant cell walls. A putative AXE gene exhibiting high homology to carbohydrate esterase family 3 was found in the genome database of the fungus Talaromyces cellulolyticus (formerly known as Acremonium cellulolyticus). A truncated form of the protein, the catalytic domain of the enzyme, was prepared and crystallized. The best crystal was obtained at 293 K using 0.17 M ammonium sulfate, 28% PEG 4000, 5%(v/v) glycerol, 0.5%(w/v) n-octyl-β-D-glucoside. X-ray diffraction data were collected to 1.50 Å resolution. The crystal belonged to space group P41212 or P43212, with unit-cell parameters a = 70.90, b = 70.90, c = 87.09 Å. One enzyme molecule per asymmetric unit gave a crystal volume per protein mass (VM) of 2.62 Å(3) Da(-1) and a solvent content of 53.0%(v/v).

Application of two newly identified and characterized feruloyl esterases from Streptomyces sp. in the enzymatic production of ferulic acid from agricultural biomass.

Ferulic acid (FA), a component of hemicellulose in plant cell walls, is a phenolic acid with several potential applications based on its antioxidant properties. Recent studies have shown that feruloyl esterase (FAE) is a key bacterial enzyme involved in FA production from agricultural biomass. In this study, we screened a library of 43 esterases from Streptomyces species and identified two enzymes, R18 and R43, that have FAE activity toward ethyl ferulate. In addition, we characterized their enzyme properties in detail. R18 and R43 showed esterase activity toward other hydroxycinnamic acid esters as well, such as methyl p-coumarate, methyl caffeate, and methyl sinapinate. The amino acid sequences of R18 and R43 were neither similar to each other, nor to other FAEs. We found that R18 and R43 individually showed the ability to produce FA from corn bran; however, combination with other Streptomyces enzymes, namely xylanase and α-l-arabinofuranosidase, increased FA production from biomass such as corn bran, defatted rice bran, and wheat bran. These results suggest that R18 and R43 are effective FAEs for the enzymatic production of FA from biomass.

Role of (1,3)(1,4)-β-Glucan in Cell Walls: Interaction with Cellulose

(1,3)(1,4)-β-D-Glucan (mixed-linkage glucan or MLG), a characteristic hemicellulose in primary cell walls of grasses, was investigated to determine both its role in cell walls and its interaction with cellulose and other cell wall polysaccharides in vitro. Binding isotherms showed that MLG adsorption onto microcrystalline cellulose is slow, irreversible, and temperature-dependent. Measurements using quartz crystal microbalance with dissipation monitoring showed that MLG adsorbed irreversibly onto amorphous regenerated cellulose, forming a thick hydrogel. Oligosaccharide profiling using endo-(1,3)(1,4)-β-glucanase indicated that there was no difference in the frequency and distribution of (1,3) and (1,4) links in bound and unbound MLG. The binding of MLG to cellulose was reduced if the cellulose samples were first treated with certain cell wall polysaccharides, such as xyloglucan and glucuronoarabinoxylan. The tethering function of MLG in cell walls was tested by applying endo-(1,3)(1,4)-β-glucanase to wall samples in a constant force extensometer. Cell wall extension was not induced, which indicates that enzyme-accessible MLG does not tether cellulose fibrils into a load-bearing network.

Functional conservation of the glycosyltransferase gene GT47A in the monocot rice.

Glucuronoarabinoxylan is the major hemicellulose in grass cell walls, yet the mechanism of xylan synthesis in monocot plants is still unclear. Unraveling the genes involved in the biosynthesis of xylan in rice will be very important for the utilization of rice straw as a source of bioenergy in the future. In this report, we investigated the functional role of a rice gene homologous to Arabidopsis IRREGULAR XYLEM10 (IRX10), belonging to the glycosyl transferase (GT) gene family 47 (GT47), in the biosynthesis of xylan. The protein sequence of OsGT47A from rice exhibits a 93.49% similarity to IRX10, which is involved in the biosynthesis of glucuronoxylan in Arabidopsis. Phylogenetic analysis of the GT47 glycosyl transferase family in the rice genome revealed that OsGT47A is a closely related homolog of IRX10 and IRX10L. Expression pattern analysis showed that the OsGT47A gene is highly expressed in the rice stem. Overexpression of OsGT47A in the irx10 irx10L double mutant rescued the plant growth phenotype and restored secondary wall thickness. Analysis of monosaccharides indicated that the rescued plants had levels of xylose identical to those of the wild type plants, and the fluorescence signals were restored in the complementation plants by xylan immunolocalization. The OsGT47A complementation under the native promoter of Arabidopsis IRX10L (ProIRX10L) partially rescued the double mutant, indicating that OsGT47A is functionally equivalent to IRX10L. Together, these results suggest that the IRX10 homolog OsGT47A exhibits functional conservation and is most likely involved in xylan synthesis in rice.

Effect of mechanical disruption on the effectiveness of three reactors used for dilute acid pretreatment of corn stover Part 1: chemical and physical substrate analysis

BackgroundThere is considerable interest in the conversion of lignocellulosic biomass to liquid fuels to provide substitutes for fossil fuels. Pretreatments, conducted to reduce biomass recalcitrance, usually remove at least some of the hemicellulose and/or lignin in cell walls. The hypothesis that led to this research was that reactor type could have a profound effect on the properties of pretreated materials and impact subsequent cellulose hydrolysis.ResultsCorn stover was dilute-acid pretreated using commercially relevant reactor types (ZipperClave® (ZC), Steam Gun (SG) and Horizontal Screw (HS)) under the same nominal conditions. Samples produced in the SG and HS achieved much higher cellulose digestibilities (88% and 95%, respectively), compared to the ZC sample (68%). Characterization, by chemical, physical, spectroscopic and electron microscopy methods, was used to gain an understanding of the effects causing the digestibility differences. Chemical differences were small; however, particle size differences appeared significant. Sum-frequency generation vibrational spectra indicated larger inter-fibrillar spacing or randomization of cellulose microfibrils in the HS sample. Simons’ staining indicated increased cellulose accessibility for the SG and HS samples. Electron microscopy showed that the SG and HS samples were more porous and fibrillated because of mechanical grinding and explosive depressurization occurring with these two reactors. These structural changes most likely permitted increased cellulose accessibility to enzymes, enhancing saccharification.ConclusionsDilute-acid pretreatment of corn stover using three different reactors under the same nominal conditions gave samples with very different digestibilities, although chemical differences in the pretreated substrates were small. The results of the physical and chemical analyses of the samples indicate that the explosive depressurization and mechanical grinding with these reactors increased enzyme accessibility. Pretreatment reactors using physical force to disrupt cell walls increase the effectiveness of the pretreatment process.

Effect of mechanical disruption on the effectiveness of three reactors used for dilute acid pretreatment of corn stover Part 2: morphological and structural substrate analysis

BackgroundLignocellulosic biomass is a renewable, naturally mass-produced form of stored solar energy. Thermochemical pretreatment processes have been developed to address the challenge of biomass recalcitrance, however the optimization, cost reduction, and scalability of these processes remain as obstacles to the adoption of biofuel production processes at the industrial scale. In this study, we demonstrate that the type of reactor in which pretreatment is carried out can profoundly alter the micro- and nanostructure of the pretreated materials and dramatically affect the subsequent efficiency, and thus cost, of enzymatic conversion of cellulose.ResultsMulti-scale microscopy and quantitative image analysis was used to investigate the impact of different biomass pretreatment reactor configurations on plant cell wall structure. We identify correlations between enzymatic digestibility and geometric descriptors derived from the image data. Corn stover feedstock was pretreated under the same nominal conditions for dilute acid pretreatment (2.0 wt% H2SO4, 160°C, 5 min) using three representative types of reactors: ZipperClave® (ZC), steam gun (SG), and horizontal screw (HS) reactors. After 96 h of enzymatic digestion, biomass treated in the SG and HS reactors achieved much higher cellulose conversions, 88% and 95%, respectively, compared to the conversion obtained using the ZC reactor (68%). Imaging at the micro- and nanoscales revealed that the superior performance of the SG and HS reactors could be explained by reduced particle size, cellular dislocation, increased surface roughness, delamination, and nanofibrillation generated within the biomass particles during pretreatment.ConclusionsIncreased cellular dislocation, surface roughness, delamination, and nanofibrillation revealed by direct observation of the micro- and nanoscale change in accessibility explains the superior performance of reactors that augment pretreatment with physical energy.

Cloning and enzymatic characterization of four thermostable fungal endo-1,4-β-xylanases

Endo-1,4-β-xylanases (EC 3.2.1.8) hydrolyze the 1,4-β-D-xylosidic linkages in xylans, the most abundant hemicellulose in plant cell walls. Xylanase enzymes have numerous industrial applications, including the manufacturing of animal feed, bread, juice and wine, pulp and paper, and biofuels. In this study, two glycosyl hydrolase family 10 members designated GtXyn10A and GtXyn10B and two glycosyl hydrolase family 11 members, OpXyn11A and CcXyn11C, were functionally expressed and subjected to biochemical characterization. The K(M), V(max), and k(cat) values of the four xylanases, determined using birchwood xylan, ranged from 0.27 to 1.1 mg/mL, 130 to 980 μmol/min/mg, and 109 to 344 s⁻¹, respectively, where OpXyn11A gave the highest and GtXyn10B the lowest values for all three parameters. Substrate specificity studies and analysis of the products released during the degradation of xylo-oligosaccharides and three types of xylan revealed significant differences in catalytic properties, particularly between OpXyn11A and the other xylanases and between the family 10 and the family 11 xylanases. Molecular modeling suggests that the unique substrate specificity of OpXyn11A can be attributed to the presence of a serine rather that an asparagine or aspartate residue at the +1 substrate binding site. Additionally, all four xylanases exhibited biochemical characteristics of interest for various commercial applications.

A Review of the Role of Amphiphiles in Biomass to Ethanol Conversion

One of the concerns for economical production of ethanol from biomass is the large volume and high cost of the cellulolytic enzymes used to convert biomass into fermentable sugars. The presence of acetyl groups in hemicellulose and lignin in plant cell walls reduces accessibility of biomass to the enzymes and makes conversion a slow process. In addition to low enzyme accessibility, a rapid deactivation of cellulases during biomass hydrolysis can be another factor contributing to the low sugar recovery. As of now, the economical reduction in lignin content of the biomass is considered a bottleneck, and raises issues for several reasons. The presence of lignin in biomass reduces the swelling of cellulose fibrils and accessibility of enzyme to carbohydrate polymers. It also causes an irreversible adsorption of the cellulolytic enzymes that prevents effective enzyme activity and recycling. Amphiphiles, such as surfactants and proteins have been found to improve enzyme activity by several mechanisms of action that are not yet fully understood. Reduction in irreversible adsorption of enzyme to non-specific sites, reduction in viscosity of liquid and surface tension and consequently reduced contact of enzyme with air-liquid interface, and modifications in biomass chemical structure are some of the benefits derived from surface active molecules. Application of some of these amphiphiles could potentially reduce the capital and operating costs of bioethanol production by reducing fermentation time and the amount of enzyme used for saccharification of biomass. In this review article, the benefit of applying amphiphiles at various stages of ethanol production (i.e., pretreatment, hydrolysis and hydrolysis-fermentation) is reviewed and the proposed mechanisms of actions are described.

Cytological and physiological basis for tomato varietal resistance againstAlternaria alternata

Since tomato is an important food component, it is imperative to enhance its yield against the activities of many devastating fungal pathogens such as Alternaria alternata. The exploitation of plant innate resistance by cultivation of resistant varieties is an effective measure in this regard. In the present study, 28 tomato varieties were tested against 32 A. alternata isolates, and representative varieties were further evaluated to determine the extent and basis of their antifungal resistance. A significant increase (104.7%) in polyphenols was recorded in the resistant variety Dinaar compared with the susceptible variety Red Tara. Dinaar also exhibited 100% enhancement of alkaloids and terpenoids along with a 30.7% increase in cell wall hemicellulose content. Significant differences were found in physical barriers (cellulose, lignin and pectin) of the representative varieties when stained tissue sections were subjected to colorimetric analysis. Similarly, polyphenol oxidase, peroxidase and phenylalanine ammonia lyase showed increases of 78.37, 114.67 and 125.11% respectively in the resistant variety. Higher expression of glucanase genes was evident from native gel analysis, in which not only the number of isozymes but also the quantity of individual isozymes was significantly increased. The resistant variety Dinaar had strong antifungal resistance and can therefore be recommended as suitable for cultivation in the agricultural system of Pakistan.

Tissue density determines the water storage characteristics of crop residues

The water storage properties of plant residues play an important role in the regulation of water retention and water transport in no-till agricultural soils. The objective of this work was to understand how the characteristics of crop residues determine their water absorption and retention properties. A range of eleven undecomposed crop residues and maize stem residue of different particle sizes at three stages of decomposition were characterized regarding their physical and chemical features. Water immersion for varying periods of time was used to determine the kinetics of water absorption and the maximal water storage for each type of residue. The immersion time required to reach an equilibrium moisture content varied greatly according to the residue type, ranging from 6 to 30 h at 20 °C. The maximal water content ranged from 1.28 to 3.81 g H2O g−1 dry residue for undecomposed residues and increased with increasing decomposition. The proportions of cellulose, hemicellulose and lignin in the plant cell walls did not explain the water storage capacities. Differences in porosity, resulting from different tissue densities and the creation of pores during decomposition, were highly correlated with differences in water storage properties. The tissue density of plant residues, which can be inferred from simple characteristics of residue mass and volume, is a relevant criterion for explaining the maximal water storage capacity of plant residues.

Purification and characterization of a xylanase from the endophytic fungus Alternaria alternata isolated from the Thai medicinal plant, Croton oblongifolius Roxb.

Xylanases are a class of enzymes that degrade b-1, 4-xylan, a linear polysaccharide found as hemicellulose in plant cell walls, in xylose. They are one of the most importantenzyme groups used in industry and agriculture. Fifty-four endophytic fungi were isolated and examined for xylanase production. Xylanase activity was found in thirty of the isolates in primary screening by growing on solid xylan agar plates. After secondary screening for xylanase activity in xylan liquid culture, the isolate that yielded the highest xylanase-production (PTRa9) was selected for further evaluation. Optimal xylanase production was achieved after 4 days of culture with 2% (w/v) rice bran and 0.1% (w/v) ammonium sulfate as the carbon and nitrogen source respectively. This xylanase was enriched 60.8-fold to apparent homogeneity by sequential ammonium sulphate precipitation, Diethylaminoethyl (DEAE)-cellulose ion exchange and Superdex 75 gel filtration chromatography. The resultant 54.8 kDa protein had a specific activity of 161.1 U/mg protein, an optimal temperature of 45°C, with >90% activity from -20 to 45°C, a broad pH range of 3.0 to 11.0 (optimal at pH 5.0), and was sensitive to most divalent cations but especially by Hg2+, Cu2+ and EDTA. From the kinetic analysis, it had a Km of 0.421 mg/ml and a Vmax of 0.826 U/mg protein. Key words: Xylanase, endophytic fungi, Thai medicinal plant.

Dynamics of Carbon Accumulation During the Fast Growth Period of Bamboo Plant

Moso bamboo (Phyllostachys pubescens) has an extremely fast growth rate, however little information is available on the dynamics of carbon accumulation during the fast growth period. Bamboo trunk were sampled at three different stages (1: shoot emergence to first shell detachment; 2: first shell detachment to branch emergence; 3: branch emergence to detachment of all shells) and divided into three parts (upper, middle and lower). The average shoot elongation rate and biomass accumulation rate were 17 cm/d and 96 g/d, respectively. The carbon content increased progressively at the growth stage, and the fixed carbon was partitioned into cell wall hemicellulose and cellulose to meet the demand of rapid cell elongation. Different rates of N, P, K, Ca, and Mg content were found among different parts. These results indicated that the fast growth of the bamboo trunk is related to the extraordinary ability of bamboo to assimilate carbon, but not consistently related to mineral nutrients absorption.

Integrative Analysis of Transgenic Alfalfa (Medicago sativa L.) Suggests New Metabolic Control Mechanisms for Monolignol Biosynthesis

The entanglement of lignin polymers with cellulose and hemicellulose in plant cell walls is a major biological barrier to the economically viable production of biofuels from woody biomass. Recent efforts of reducing this recalcitrance with transgenic techniques have been showing promise for ameliorating or even obviating the need for costly pretreatments that are otherwise required to remove lignin from cellulose and hemicelluloses. At the same time, genetic manipulations of lignin biosynthetic enzymes have sometimes yielded unforeseen consequences on lignin composition, thus raising the question of whether the current understanding of the pathway is indeed correct. To address this question systemically, we developed and applied a novel modeling approach that, instead of analyzing the pathway within a single target context, permits a comprehensive, simultaneous investigation of different datasets in wild type and transgenic plants. Specifically, the proposed approach combines static flux-based analysis with a Monte Carlo simulation in which very many randomly chosen sets of parameter values are evaluated against kinetic models of lignin biosynthesis in different stem internodes of wild type and lignin-modified alfalfa plants. In addition to four new postulates that address the reversibility of some key reactions, the modeling effort led to two novel postulates regarding the control of the lignin biosynthetic pathway. The first posits functionally independent pathways toward the synthesis of different lignin monomers, while the second postulate proposes a novel feedforward regulatory mechanism. Subsequent laboratory experiments have identified the signaling molecule salicylic acid as a potential mediator of the postulated control mechanism. Overall, the results demonstrate that mathematical modeling can be a valuable complement to conventional transgenic approaches and that it can provide biological insights that are otherwise difficult to obtain.

The DUF579 domain containing proteins IRX15 and IRX15‐L affect xylan synthesis in Arabidopsis

Xylan is the principal hemicellulose in the secondary cell walls of eudicots and in the primary and secondary cell walls of grasses and cereals. The biosynthesis of this important cell wall component has yet to be fully determined although a number of proteins have been shown to be required for xylan synthesis. To discover new genes involved in xylan biosynthesis we explored the psyllium (Plantago ovata Forsk) seed mucilaginous layer through EST profiling. This tissue synthesizes large amounts of a complex heteroxylan over a short period of time. By comparing abundant transcripts in this tissue with abundant transcripts specifically present during secondary cell wall formation in Arabidopsis thaliana, where glucuronoxylan biosynthesis is pronounced, we identified two Arabidopsis genes likely involved in xylan biosynthesis. These genes encode proteins containing a Domain of Unknown Function (DUF) 579 and were designated IRREGULAR XYLEM (IRX) 15 and IRX15-LIKE (IRX15-L). We obtained Arabidopsis T-DNA knockout lines for the two genes and analyzed their lower stems for changes in neutral monosaccharide composition. No changes were observed in each of these mutants, although the irx15 irx15-L double mutant displayed a moderate reduction in stem xylose. Further characterization of the irx15 irx15-L mutant revealed irregular secondary cell wall margins in fiber cells and a lower xylan degree of polymerization. Through these studies we conclude that IRX15 and IRX15-L function in a redundant manner and are involved in xylan biosynthesis.

Cell wall hemicellulose contributes significantly to aluminum adsorption and root growth in Arabidopsis.

The cell wall (CW) has been recognized as the major target of aluminum (Al) toxicity. However, the components responsible for Al accumulation and the mechanisms of Al-induced CW function disruption are still elusive. The contribution of different CW components (pectin, hemicellulose 1 [HC1], and HC2) to adsorb Al and the effect of Al on xyloglucan endotransglucosylase/hydrolyase activity were investigated in Arabidopsis (Arabidopsis thaliana) in this study. A fractionation procedure was optimized to effectively extract different CW components, especially to prevent the HC fraction from pectin contamination. When CW materials extracted from Al-treated roots (50 μm Al for 24 h) were fractionated, about 75% of CW Al accumulated in the HC1 fraction. A time-dependent kinetic study showed that only when the HC1 fraction was removed was the amount of Al adsorbed decreased sharply. In vivo localization of xyloglucan endotransglucosylase (XET) activity showed that Al greatly inhibited this enzyme activity within 30 min of exposure, which was concomitant with Al-induced callose deposition in roots. Results from real-time reverse transcription-polymerase chain reaction indicated that three genes may constitute the major contributors to XET activity and that the inhibition of XET activity by Al is caused by transcriptional regulation. These results, to our knowledge for the first time, demonstrate that HC is the major pool for Al accumulation. Furthermore, Al-induced reduction in XET activity could play an important role in Al-induced root growth inhibition.