Variation In Cell Wall Composition Research Articles

Nanoceria-induced variations in leaf anatomy and cell wall composition drive the increase in mesophyll conductance of salt-stressed cotton leaves

Nanomaterials as an emerging tool are being used to improve plant's net photosynthetic rate (AN) when suffering salt stress, but the underlying mechanisms remain unclear. To clarify this, a hydroponic experiment was conducted to study the effects of polyacrylic acid coated nanoceria (PNC) on the AN of salt-stressed cotton and related intrinsic mechanisms. Results showed that the PNC-induced AN enhancement of salt-stressed leaves was strongly facilitated by the mesophyll conductance to CO2 (gm). Further analysis showed that the PNC-induced improvement of gm was related to the increased chloroplast surface area exposed to intercellular airspaces, which was attribute to the increased mesophyll surface area exposed to intercellular airspaces and chloroplast number due to the increased K+ content and decreased reactive oxygen species level in salt-stressed leaves. Interestingly, our results also showed that PNC-induced variations in cell wall composition of salt-stressed cotton leaves strongly influenced gm, especially, hemicellulose and pectin. Moreover, the proportion of pectin in cell wall composition played a more important role in determining gm. Our study demonstrated for the first time that nanoceria, through alterations to anatomical traits and cell wall composition, drove gm enhancement, which ultimately increased AN of salt-stressed leaves.

Seasonal Variation in Cell Wall Composition and Carbohydrate Metabolism in the Seagrass Posidonia oceanica Growing at Different Depths.

Posidonia oceanica is a common seagrass in the Mediterranean Sea that is able to sequester large amounts of carbon. The carbon assimilated during photosynthesis can be partitioned into non-structural sugars and cell-wall polymers. In this study, we investigated the distribution of carbon in starch, soluble carbohydrates and cell-wall polymers in leaves and rhizomes of P. oceanica. Analyses were performed during summer and winter in meadows located south of the Frioul archipelago near Marseille, France. The leaves and rhizomes were isolated from plants collected in shallow (2 m) and deep water (26 m). Our results showed that P. oceanica stores more carbon as starch, sucrose and cellulose in summer and that this is more pronounced in rhizomes from deep-water plants. In winter, the reduction in photoassimilates was correlated with a lower cellulose content, compensated with a greater lignin content, except in rhizomes from deep-water plants. The syringyl-to-guaiacyl (S/G) ratio in the lignin was higher in leaves than in rhizomes and decreased in rhizomes in winter, indicating a change in the distribution or structure of the lignin. These combined data show that deep-water plants store more carbon during summer, while in winter the shallow- and deep-water plants displayed a different cell wall composition reflecting their environment.

Characterization of the nanomechanical properties of the fission yeast (Schizosaccharomyces pombe) cell surface by atomic force microscopy.

Variations in cell wall composition and biomechanical properties can contribute to the cellular plasticity required during complex processes such as polarized growth and elongation in microbial cells. This study utilizes atomic force microscopy (AFM) to map the cell surface topography of fission yeast, Schizosaccharomyces pombe, at the pole regions and to characterize the biophysical properties within these regions under physiological, hydrated conditions. High‐resolution images acquired from AFM topographic scanning reveal decreased surface roughness at the cell poles. Force extension curves acquired by nanoindentation probing with AFM cantilever tips under low applied force revealed increased cell wall deformation and decreased cellular stiffness (cellular spring constant) at cell poles (17 ± 4 mN/m) relative to the main body of the cell that is not undergoing growth and expansion (44 ± 10 mN/m). These findings suggest that the increased deformation and decreased stiffness at regions of polarized growth at fission yeast cell poles provide the plasticity necessary for cellular extension. This study provides a direct biophysical characterization of the S. pombe cell surface by AFM, and it provides a foundation for future investigation of how the surface topography and local nanomechanical properties vary during different cellular processes.

Multivariate Raman mapping for phenotypic characterization in plant tissue sections

Identifying and characterizing the biochemical variation in plant tissues is an important task in many research fields. Small spectral differences of the plant cell wall that are caused by genetic or environmental influences may be superimposed by individual variation as well as by a microscopic heterogeneity in molecular composition and structure of different histological substructures. A set of 56 samples from Cucumis sativus (cucumber) plants, comprising a total of ~168,000 spectra from tissue sections of leaf, stem, and roots was investigated by Raman microspectroscopic mapping excited at 532 nm. A multivariate analysis was carried out in order to assess the variation of the spectra with respect to origin of the tissue, the histological (cell wall) substructures, and the possibility to discriminate the spectra obtained from different individuals that had been subjected to two different conditions during growth. Combining the results of principal component analysis (PCA) based classification with the original spatial information in the maps of 23 sections of leaf xylem, variation in cell wall composition is found for four different individuals that also includes a discrimination of tissue grown in the presence and absence of additional silicic acid in the irrigation water of the plants. The spectral data point to differences in a contribution by carotenoids, as well as by hydroxycinnamic acids to the spectra. The results give new insight into the chemical heterogeneity of plant tissues and may be useful for elucidating biochemical processes associated with biomineralization by vibrational spectroscopy.

Cell wall composition strongly influences mesophyll conductance in gymnosperms.

Cell wall thickness is widely recognized as one of the main determinants of mesophyll conductance to CO2 (gm ). However, little is known about the components that regulate effective CO2 diffusivity in the cell wall (i.e. the ratio between actual porosity and tortuosity, the other two biophysical diffusion properties of cell walls). The aim of this study was to assess, at the interspecific level, potential relationships between cell wall composition, cell wall thickness (Tcw ) and gm . Gymnosperms constitute an ideal group to deepen these relationships, as they present, on average, the thickest cell walls within spermatophytes. We characterized the foliar gas exchange, the morphoanatomical traits related with gm , the leaf fraction constituted by cell walls and three main components of primary cell walls (hemicelluloses, cellulose and pectins) in seven gymnosperm species. We found that, although the relatively low gm of gymnosperms was mainly determined by their elevated Tcw , gm was also strongly correlated with cell wall composition, which presumably sets the final effective CO2 diffusivity. The data presented here suggest that (i) differences in gm are strongly correlated to the pectins to hemicelluloses and cellulose ratio in gymnosperms, and (ii) variations in cell wall composition may modify effective CO2 diffusivity in the cell wall to compensate the negative impact of thickened walls. We speculate that higher relative pectin content allows higher gm because pectins increase cell wall hydrophilicity and CO2 molecules cross the wall dissolved in water.

The toxicity of graphene oxide affected by algal physiological characteristics: A comparative study in cyanobacterial, green algae, diatom

Though the main toxic mechanisms of graphene oxide (GO) to algae have been accepted as the shading effect, oxidative stress and mechanical damage, the effect of algal characteristics on these three mechanisms of GO toxicity have seldom been taken into consideration. In this study, we investigated GO toxicity to green algae (Chlorella vulgaris, Scenedesmus obliquus, Chlamydomonas reinhardtii), cyanobacteria (Microcystis aeruginosa) and diatoms (Cyclotella sp.). The aim was to assess how the physiological characteristics of algae affect the toxicity of GO. Results showed that 10 mg/L of GO significantly inhibited the growth of all tested algal types, while S. obliquus and C. reinhardtii were found to be the most susceptible and tolerant species, respectively. Then, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the physiological characteristics of the assessed algae. The presence of locomotive organelles, along with smaller and more spherical cells, was more likely to alleviate the shading effect. Variations in cell wall composition led to different extents of mechanical damage as shown by Cyclotella sp. silica frustules and S. obliquus autosporine division being prone to damage. Meanwhile, growth inhibition and cell division were significantly correlated with the oxidative stress and membrane permeability, suggesting the latter two indicators can effectively signal GO toxicity to algae. The findings of this study provide novel insights into the toxicity of graphene materials in aquatic environments.

Phenotypic Variation of Cell Wall Composition and Stem Morphology in Hemp (Cannabis sativa L.): Optimization of Methods.

The growing demands for sustainable fibers have stimulated the study of genetic diversity in the quality of hemp fiber (Cannabis sativa L.). Nevertheless, the lack of high-throughput phenotyping methods that are suited for the analysis of hemp fiber, hampers the analysis of many accessions, and consequently the breeding for this complex trait. In the present report, we developed and optimized the throughput of five methods to study the diversity in hemp fiber quality including cell wall extraction, biochemical composition of cell wall polysaccharides, quantification of lignin, quantification of crystalline polysaccharides and morphology of the stems. Six hemp accessions contrasting for cell wall properties were used to assess the throughput and suitability of these methods for genetic studies. The methods presented revealed to be highly repeatable, with low coefficients of variation between technical replicates. With these methods we were able to detect significant phenotypic variation in cell wall composition and stem morphology between the six accessions. In addition, the throughput of the methods has been upgraded to a level that enables their use for phenotyping cell wall traits in breeding programs. The cell wall extraction was optimized to extract enough material for the complete characterization of the cell wall of hemp while reducing the time for the entire analysis. The throughput of the stem morphological analysis was improved by decreasing the timing of fixation, infiltration, and embedding of mature and dry hemp stems. Notwithstanding, our methods already have the potential to phenotype large number of accessions in a relatively short period of time. Our methods will enable exploration of genetic diversity of fiber quality and will contribute to the development of new hemp varieties with advanced quality of fibers.

Isolation and evaluation of a novel strain of Chlorella sorokiniana that resists grazing by the predator Poterioochromonas malhamensis

The protozoan flagellate Poterioochromonas malhamensis is one of the main contaminants in Chlorella culture; however, few effective and affordable methods have been developed to control it. Unexpectedly, one strain of an unknown green alga was observed to be capable of contaminating P. malhamensis culture and defending itself against the protozoan. Based on cell morphology and molecular information, the green microalga was identified as Chlorella sorokiniana CMBB-146. Feeding experiments showed that the addition of the predator P. malhamensis not only had no negative effect on the growth of C. sorokiniana CMBB-146, but also stimulated an increase in the biomass of C. sorokiniana CMBB-146. The grazing resistance of C. sorokiniana CMBB-146 against P. malhamensis was also verified on a pilot scale with 100-L raceway ponds. Further experiments revealed that P. malhamensis showed a strong ability in ingesting C. sorokiniana CMBB-146 cells, but the ingested cells were hard to digest. Through comparison with other species of Chlorella without grazing resistance, we found that the differences on cell size, cell morphology, and biochemical composition had no relation with the ability of C. sorokiniana CMBB-146 to resist grazing by P. malhamensis. Finally, it is speculated that variation in cell wall composition, especially the content of galactosamine, empowers C. sorokiniana CMBB-146 to resist P. malhamensis. Therefore, our study confirms that C. sorokiniana CMBB-146 is a strong candidate as a potential production organism for outdoor massive cultivation, and as a potential microalgal strain for studying how microalgae resist grazing by protozoa.

Genetic and environmental factors contribute to variation in cell wall composition in mature desi chickpea (Cicer arietinum L.) cotyledons.

Chickpea (Cicer arietinum L.) is an important nutritionally rich legume crop that is consumed worldwide. Prior to cooking, desi chickpea seeds are most oftendehulled and cleaved to release the split cotyledons, referred to as dhal. Compositional variation between desi genotypes has a significant impact on nutritional quality and downstream processing, and this has been investigated mainly in terms of starch and protein content. Studies in pulses such as bean and lupin have also implicated cell wall polysaccharides in cooking time variation, but the underlying relationship between desi chickpea cotyledon composition and cooking performance remains unclear. Here, we utilized a variety of chemical and immunohistological assays to examine details of polysaccharide composition, structure, abundance, and location within the desi chickpea cotyledon. Pectic polysaccharides were the most abundant cell wall components, and differences in monosaccharide and glycosidic linkage content suggest both environmental and genetic factors contribute to cotyledon composition. Genotype-specific differences were identified in arabinan structure, pectin methylesterification, and calcium-mediated pectin dimerization. These differences were replicated in distinct field sites and suggest a potentially important role for cell wall polysaccharides and their underlying regulatory machinery in the control of cooking time in chickpea.

The application of Fourier transform mid-infrared (FTIR) spectroscopy to identify variation in cell wall composition of Setaria italica ecotypes

Cell wall composition in monocotyledonous grasses has been identified as a key area of research for developing better feedstocks for forage and biofuel production. Setaria viridis and its close domesticated relative Setaria italica have been chosen as suitable monocotyledonous models for plants possessing the C4 pathway of photosynthesis including sorghum, maize, sugarcane, switchgrass and Miscanthus×giganteus. Accurate partial least squares regression (PLSR) models to predict S. italica stem composition have been generated, based upon Fourier transform mid-infrared (FTIR) spectra and calibrated with wet chemistry determinations of ground S. italica stem material measured using a modified version of the US National Renewable Energy Laboratory (NREL) acid hydrolysis protocol. The models facilitated a high-throughput screening analysis for glucan, xylan, Klason lignin and acid soluble lignin (ASL) in a collection of 183 natural S. italica variants and clustered them into classes, some possessing unique chemotypes. The predictive models provide a highly efficient screening tool for large scale breeding programs aimed at identifying lines or mutants possessing unique cell wall chemotypes. Genes encoding key catalytic enzymes of the lignin biosynthesis pathway exhibit a high level of conservation with matching expression profiles, measured by RT-qPCR, among accessions of S. italica, which closely mirror profiles observed in the different developmental regions of an elongating internode of S. viridis by RNASeq.

Contributions of a unique β-clamp to substrate recognition illuminates the molecular basis of exolysis in ferulic acid esterases.

Lignocellulosic biomass is a promising renewable resource; however, deconstruction of this material is still the rate-limiting step. Major obstacles in the biocatalytic turnover of lignocellulose are ester-linked decorations that prevent access to primary structural polysaccharides. Enzymes targeting these esters represent promising biotools for increasing bioconversion efficiency. Ruminant livestock are unique in their ability to degrade lignocellulose through the action of their gut microbiome. The anaerobic fungi (phylum Neocallimastigomycota) are key members of this ecosystem that express a large repertoire of carbohydrate-active enzymes (CAZymes) with little sequence identity with characterized CAZymes [Lombard, Golaconda, Drula, Coutinho and Henrissat (2014) Nucleic Acids Res. 42: , D490-D495]. We have identified a carbohydrate esterase family 1 (CE1) ferulic acid esterase (FAE) belonging to Anaeromyces mucronatus(AmCE1/Fae1a), and determined its X-ray structure in both the presence [1.55 Å (1 Å=0.1 nm)] and absence (1.60 Å) of ferulic acid. AmCE1 adopts an α/β-hydrolase fold that is structurally conserved with bacterial FAEs, and possesses a unique loop, termed the β-clamp, that encloses the ligand. Isothermal titration calorimetry reveals that substrate binding is driven by enthalpic contributions, which overcomes a large entropic penalty. A comparative analysis of AmCE1 with related enzymes has uncovered the apparent structural basis for differential FAE activities targeting cross-linking ferulic acid conjugates compared with terminal decorations. Based on comparisons to structurally characterized FAEs, we propose that the β-clamp may define the structural basis of exolytic activities in FAEs. This provides a structure-based tool for predicting exolysis and endolysis in CE1. These insights hold promise for rationally identifying enzymes tailored for bioconversion of biomass with variations in cell wall composition.

Diverse cell wall composition and varied biomass digestibility in wheat straw for bioenergy feedstock

Wheat straw has a vast potential as feedstock for biofuel production in China. However, little information is available regarding variation in cell wall composition and enzymatic digestibility of wheat straw. This study investigated cell wall compositions and biomass digestibility of the straw of 115 wheat accessions from central China using a 1% NaOH pretreatment and mixed enzymatic hydrolysis. Significant variation in cell wall composition and sugar release was observed, with a coefficient variation (CV) ranging from 4.7% to 21.2%. Cellulose, hemicelluloses, alkali detergent hemicelluloses (ADH), and acid insoluble lignin (AIL) positively correlated with each other, and they all negatively correlated with acid soluble lignin (ASL). Hexose yields had a negative correlation with ADH and AIL, and positive correlation with ASL. No apparent undesirable correlation was found between sugar release and grain yield, thus yield and biomass convertibility can be potentially improved simultaneously. Furthermore, the features of the cell wall constitutions were compared with other plants and their implication in determining the best possible conversion strategy was discussed. This initial study is essential to understand the cell wall composition of wheat straw and to explore the potential of wheat straw as feedstock for biofuel production.

Developmental anatomy and immunocytochemistry reveal the neo-ontogenesis of the leaf tissues of Psidium myrtoides (Myrtaceae) towards the globoid galls of Nothotrioza myrtoidis (Triozidae).

The temporal balance between hyperplasia and hypertrophy, and the new functions of different cell lineages led to cell transformations in a centrifugal gradient that determines the gall globoid shape. Plant galls develop by the redifferentiation of new cell types originated from those of the host plants, with new functional and structural designs related to the composition of cell walls and cell contents. Variations in cell wall composition have just started to be explored with the perspective of gall development, and are herein related to the histochemical gradients previously detected on Psidium myrtoides galls. Young and mature leaves of P. myrtoides and galls of Nothotrioza myrtoidis at different developmental stages were analysed using anatomical, cytometrical and immunocytochemical approaches. The gall parenchyma presents transformations in the size and shape of the cells in distinct tissue layers, and variations of pectin and protein domains in cell walls. The temporal balance between tissue hyperplasia and cell hypertrophy, and the new functions of different cell lineages led to cell transformations in a centrifugal gradient, which determines the globoid shape of the gall. The distribution of cell wall epitopes affected cell wall flexibility and rigidity, towards gall maturation. By senescence, it provided functional stability for the outer cortical parenchyma. The detection of the demethylesterified homogalacturonans (HGAs) denoted the activity of the pectin methylesterases (PMEs) during the senescent phase, and was a novel time-based detection linked to the increased rigidity of the cell walls, and to the gall opening. Current investigation firstly reports the influence of immunocytochemistry of plant cell walls over the development of leaf tissues, determining their neo-ontogenesis towards a new phenotype, i.e., the globoid gall morphotype.

Transcriptome analysis and gene expression atlas for Panicum hallii var. filipes, a diploid model for biofuel research

Panicum hallii is an emerging model for genetic studies of agronomic traits in Panicum, presenting a tractable diploid alternative study system to the tetra- or octaploid biofuel crop switchgrass (Panicum virgatum). To characterize the gene complement in P.hallii var. filipes and enable gene expression analysis in this system we sequenced, assembled, and annotated the transcriptome. Over 300 Mb of normalized cDNA prepared from multiple tissues and treatments was sequenced using 454-Titanium, producing an annotated assembly including 15 422 unique gene names. Comparison with other grass genomes identified putative P.hallii homologs for >14 000 previously characterized genes. We also developed an atlas of gene expression across tissues and stages using RNA-Seq (the quantitative analysis of short cDNA reads). SOLiD sequencing and quantitative analysis of more than 40 million cDNA tags identified substantial variation in expression profiles among tissues, consistent with known functional differences. Putative homologs were found for all enzymes in the phenylpropanoid pathway leading to lignin biosynthesis, including genes with known effects on biomass conversion efficiency. The resources developed here will enable studies of the genes underlying variation in cell wall composition, drought tolerance, and biomass production in Panicum.

Variation in Cell Wall Composition and Accessibility in Relation to Biofuel Potential of Short Rotation Coppice Willows

Short rotation coppice (SRC) willow is currently emerging as an important dedicated lignocellulosic energy crop in the UK. However, investigation into the variation between species and genotypes in their suitability for liquid transport biofuel processing has been limited. To address this, four traits relevant to biofuel processing (composition, enzymatic saccharification, response to pretreatment and projected ethanol yields) were studied in 35 genotypes of willow including Europe’s leading SRC willow cultivars. Large, genotype-specific variation was observed for all four traits. Significant positive correlations were identified between the accessibility of glucan to enzymatic saccharification before and after pretreatment as well as glucose release and xylose release via acid hydrolysis during pretreatment. Of particular interest is that the lignin content of the biomass did not correlate with accessibility of glucan to enzymatic saccharification. The genotype-specific variations identified have implications for SRC willow breeding and for potential reductions in both the net energy expenditure and environmental impact of the lignocellulosic biofuel process chain. The large range of projected ethanol yields demonstrate the importance of feedstock selection based on an ideotype encompassing the performance of both field biomass growth and ease of conversion.

Genotypic variation in cell wall composition in a diverse set of 244 accessions of Miscanthus

'Biofuels and Bioenergy: Challenges and Opportunities’ Proceedings of a joint workshop of IEA Bioenergy Tasks 29, 31 and 39, August 2006, Vancouver, British Columbia, Canada IMPF: 03.64 RONO: 03134;;06339 Sponsorship: BBSRC (Bioenergy & Biorenewables ISPG), EPSRC (Supergen-Bioenergy, GR/S28204) and DEFRA Sponsors: BBSRC RONO: BBS/E/W/00003134C

Genotypic and environmentally derived variation in the cell wall composition of Miscanthus in relation to its use as a biomass feedstock

Fifteen Miscanthus genotypes grown in five locations across Europe were analysed to investigate the influence of genetic and environmental factors on cell wall composition. Chemometric techniques combining near infrared reflectance spectroscopy (NIRS) and conventional chemical analyses were used to construct calibration models for determination of acid detergent lignin (ADL), acid detergent fibre (ADF), and neutral detergent fibre (NDF) from sample spectra. Results generated were subsequently converted to lignin, cellulose and hemicellulose content and used to assess the genetic and environmental variation in cell wall composition of Miscanthus and to identify genotypes which display quality traits suitable for exploitation in a range of energy conversion systems. The NIRS calibration models developed were found to predict concentrations with a good degree of accuracy based on the coefficient of determination (R2), standard error of calibration (SEC), and standard error of cross-validation (SECV) values. Across all sites mean lignin, cellulose and hemicellulose values in the winter harvest ranged from 76–115 g kg-1, 412–529 g kg-1, and 235–338 g kg-1 respectively. Overall, of the 15 genotypes Miscanthus x giganteus and Miscanthus sacchariflorus contained higher lignin and cellulose concentrations in the winter harvest. The degree of observed genotypic variation in cell wall composition indicates good potential for plant breeding and matching feedstocks to be optimised to different energy conversion processes.

Variation in cell wall composition among forage maize (Zea mays L.) inbred lines and its impact on digestibility: analysis of neutral detergent fiber composition by pyrolysis-gas chromatography-mass spectrometry.

Cell wall digestibility is an important determinant of forage quality, but the relationship between cell wall composition and digestibility is poorly understood. We analyzed the neutral detergent fiber (NDF) fraction of nine maize inbred lines and one brown midrib3 mutant with pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). Among 29 pyrolysis fragments that were quantified, two carbohydrate-derived and six lignin-derived fragments showed statistically significant genetic variation. The pyrolysis products 4-vinyl phenol and 2,6-dimethoxy-4-vinyl phenol were negatively correlated with digestibility, whereas furfural and 3-(4-hydroxyphenyl)-3-oxopropanal showed a positive correlation with digestibility. Linear discriminant analysis of the pyrolysis data resulted in the resolution of groups of inbred lines with different digestibility properties based on their chemical composition. These analyses reveal that digestibility is governed by complex interactions between different cell wall compounds, but that several pyrolysis fragments can be used as markers to distinguish between maize lines with different digestibility.

Screening of Arabidopsis thaliana stems for variation in cell wall polysaccharides

A high-throughput method is described by which Arabidopsis thaliana stems can be screened for variation in cell wall composition after hydrolysis with Driselase or trifluoroacetic acid (TFA). Driselase, a mixture of fungal enzymes, hydrolyses cellulose (to glucose) and all the major matrix polysaccharides (to monosaccharides and/or characteristic disaccharides); TFA hydrolyses the matrix polysaccharides, but not cellulose, to monosaccharides. Two different wild-type ecotypes, Columbia and Wassilewskija, showed only minor differences in wall carbohydrate composition. A small number of T-DNA-tagged populations that were screened contained individuals in which the proportion of cellulose, xyloglucan or xylan differed quantitatively from the wild-type. Differences from the wild-type were also observed in the susceptibility of the hemicelluloses to hydrolysis by Driselase, probably reflecting differences in wall architecture.

A rapid mechanical lysing procedure for routine analysis of plasmids from lactobacilli, isolated from sourdoughs

Variations in cell wall composition of lactobacilli complicate attempts to standardize plasmid detection on the basis of enzymatic lysing procedures. Mechanical disruption of cell walls by a horizontal mixer-mill proved to be more effective, rapid and reproducible. In order to judge the risk of damaging plasmid deoxyribonucleic acids (DNA) in this way, plasmid profiles obtained after enzymatic lysing and mechanical disruption of cell walls were compared. As no differences were detected the mechanical procedure was accepted as a suitable method for routine analysis of plasmid profiles.