Algal Cell Wall Research Articles

Activation of chicken gamma-delta T lymphocytes by a purified ulvan extract

Chicken γδ T lymphocytes are present in a variety of tissues such as blood, spleen and intestine. They constitute a major cytotoxic population. In chicken, Salmonella immunization as well as vaccination against Newcastle disease virus are accompanied by an increase of γδ T lymphocytes in peripheral blood, which may be activated, and thus represent a protective immune response. It has been published that activation of avian γδ T cells can occur in a MHC non-restricted manner. Ulvans are complex sulfated polysaccharides composed of disaccharide repetitions found in the cell walls of green algae belonging to the genus Ulva. We recently demonstrated that a purified ulvan extract activates chicken heterophils and monocytes in vivo through TLR2 and TLR4 receptors when given in drinking water. We demonstrate here, that the same extract given once in drinking water at 25 and 50 mg/l, results in increased membrane expression of Major Histocompatibility Complex class 2 as soon as day 2, as detected using flow cytometry. We conclude chicken γδ T lymphocytes to be activated, or at least primed, in vivo, with the extract. Further experiments are required to fully understand whether their activation or priming is the result of direct and/or indirect mechanisms.

Molecular cloning and characterization of a thermostable and halotolerant endo-β-1,4-glucanase from Microbulbifer sp. ALW1.

The bacterium Microbulbifer sp. ALW1 was previously characterized with the capability to break down the cell wall of brown algae into fine pieces. The biological functions of strain ALW1 were yet to be elucidated. In this study, a gene, namely MaCel5A, was isolated from the ALW1 strain genome, encoding an endo-β-1,4-glucanase. MaCel5A was phylogenetically categorized under the glycoside hydrolase family GH5, with the highest identity to a putative cellulase of Microbulbifer thermotolerans. The recombinant MaCel5A protein purified from heterologous expression in E. coli exhibited maximum activity at 50°C and pH 6.0, respectively, and functioned selectively toward carboxymethyl cellulose and barley β-glucan. Recombinant MaCel5A demonstrated considerable tolerance to the exposure to high temperature up to 80°C for 30min retaining 49% residual activity. In addition, MaCel5A showed moderate stability against pH 5.0-11.0 and strong stability in the presence of nonionic surfactant. MaCel5A exhibited strong halo-stability and halotolerance. The activity of the enzyme increased about tenfold at 0.5M NaCl, and about fivefold even at 4.0M NaCl compared to the enzyme activity without the addition of salt. The two conserved glutamic acid residues in MaCel5A featured the typical catalytic acid/base and nucleophile machinery of glycoside hydrolases. These characteristics highlight the industrial application potential of MaCel5A.

The ciliate Paramecium bursaria allows budding of symbiotic Chlorella variabilis cells singly from the digestive vacuole membrane into the cytoplasm during algal reinfection.

The ciliate Paramecium bursaria harbors several hundred symbiotic Chlorella spp. cells in the cytoplasm. Algal re-endosymbiosis can be artificially induced using alga-removed P. bursaria. During algal re-endosymbiosis, algae ingested into the host digestive vacuoles (DVs) avoid digestion by the host lysosomal enzymes and then escape into the cytoplasm by budding off of the DV membrane. The budded alga-enclosing DV membrane then differentiates into the symbiosome or perialgal vacuole (PV) membrane and is localized beneath the host cell cortex. In this study, we determined whether the PV membrane has the ability to recognize the symbiotic alga singly by eliminating other small microspheres in the same DV. To clarify the accuracy of the budding process, we mixed fluorescent-labeled microspheres of diameter 0.20µm with isolated symbiotic algae during algal re-endosymbiosis. No fluorescence was observed from the PV membrane, as expected, and the budding DVs that enclosed both undigested and digested algae. Additionally, the algal re-endosymbiosis rate was significantly reduced in the presence of microspheres. These observations showed that the host P. bursaria allowed budding of the algae singly from the membranes of DVs without microspheres and this process required close contact between the DV membrane and the algal cell wall.

Algal Lysis by Sagittula stellata for the Production of Intracellular Valuables.

The purpose of this study was to examine the efficacy of the algicidal bacterium Sagittula stellata on the cell lysis of Nannochloropsis oceanica, a microalga found in the marine environment, in order to extract intracellular valuables. Algicidal bacteria are capable of lysing algal cell walls while keeping lipids and proteins intact yet separated. We obtained these microbes from locations with consistent algae blooms and found that the bacterium Sagittula stellata displayed significant algicidal properties toward Nannochloropsis oceanica, achieving an algicidal rate of 80.1%. We detected a decrease of 66.2% in in vivo fluorescence intensity in algae cultures, obtained a recoverable crude lipid content of 23.3% and a polyunsaturated fatty acid (PUFA) ratio of 29.0% of bacteria-treated algae, and observed the lysis of the cell membrane and the structure of the nucleus of algae. We also identified the inhibited transcription of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS) gene and proliferating cell nuclear antigen (PCNA)-related genes and the upregulated heat shock protein (hsp) gene in algal cells during bacterial exposure. Our results indicate that Sagittula stellata effectively lysed microalgae cells, allowing the recovery of intracellular valuables. The algicidal method of Sagittula stellata on Nannochloropsis oceanica cells was confirmed to be a direct attack (or predation), followed by an indirect attack through the secretion of extracellular algicidal compounds. This study provides an important framework for the broad application of algicidal microorganisms in algal cell disruption and the production of intracellular valuables.

Synthetic Biomimetic Polymethacrylates: Promising Platform for the Design of Anti-Cyanobacterial and Anti-Algal Agents.

Extensive, uncontrolled growth of algae and cyanobacteria is an environmental, public health, economic, and technical issue in managing natural and engineered water systems. Synthetic biomimetic polymers have been almost exclusively considered antimicrobial alternatives to conventional antibiotics to treat human bacterial infections. Very little is known about their applicability in an aquatic environment. Here, we introduce synthetic biomimetic polymethacrylates (SBPs) as a cost-effective and chemically facile, flexible platform for designing a new type of agent suitable for controlling and mitigating photosynthetic microorganisms. Since SBPs are cationic and membranolytic in heterotrophic bacteria, we hypothesized they could also interact with negatively charged cyanobacterial or algal cell walls and membranes. We demonstrated that SBPs inhibited the growth of aquatic photosynthetic organisms of concern, i.e., cyanobacteria (Microcystis aeruginosa and Synechococcus elongatus) and green algae (Chlamydomonas reinhardtii and Desmodesmus quadricauda), with 50% effective growth-inhibiting concentrations ranging between 95 nM and 6.5 μM. Additionally, SBPs exhibited algicidal effects on C. reinhardtii and cyanocidal effects on picocyanobacterium S. elongatus and microcystin-producing cyanobacterium M. aeruginosa. SBP copolymers, particularly those with moderate hydrophobic content, induced more potent cyanostatic and cyanocidal effects than homopolymers. Thus, biomimetic polymers are a promising platform for the design of anti-cyanobacterial and anti-algal agents for water treatment.

Enhancement of Biogas Production from Macroalgae Ulva latuca via Ozonation Pretreatment

One of the dominant species of green algae growing along the Mediterranean coast of Egypt is Ulva lactuca. Pretreatment can have a major effect on biogas production because hydrolysis of the algae cell wall structure is a rate-limiting stage in the anaerobic digestion (AD) process. The use of ozone, a new pretreatment, to boost biogas production from the green algae Ulva lactuca was investigated in this study. Ozonation at various dosages was used in contrast to untreated biomass, and the effect on the performance of subsequent mesophilic AD using two separate inoculums (cow manure and activated sludge) was examined. The findings indicated that, in different studies, ozonation pretreatment showed a substantial increase in biogas yield relative to untreated algae. With an ozone dose of 249 mg O3 g–1 VS algal for Ulva lactuca, the highest biogas output (498.75 mL/g VS) was achieved using cow manure inoculum. The evaluation of FTIR, TGA, SEM, and XRD revealed the impact of O3 on the structure of the algal cell wall and integrity breakage, which was thus established as the main contributor to improving the biogas production.

Current prospects and future developments in algal bio-hydrogen production: a review

The use of fossil fuels for energy demands is insufficient for escalating demands as they are limited in nature and are not environment friendly. Various microbial sources are under scanner for clean energy production out of which algae comes as a promising one. Production of biohydrogen from algae has brought a lot of hopes in the energy production sector. Algae are powerhouse of renewable chemicals including biofuel precursors. Biohydrogen is one of such biofuel precursors; it proves to be a clean and sustainable fuel and can be produced in industries. Currently however, it cannot stand up to the present energy demands of the world due to certain obstacles, such as cost effectiveness, storage and transportation of hydrogen. One of the main hurdles in algal biofuel technology is the rigid nature of algal cell wall. To get around this, pretreatment of algal biomass is imperative to overcome productivity issues, so as not to compromise biofuel yield. Improving the biofuel production at every step can make a huge difference in outcomes and thus comes up as a promising tool. Therefore, in the present state of art review, various methods of algal biomass pretreatment, different enzymes involved in hydrogen production, various factors influencing hydrogen production from algae and genetic engineering avenues have been discussed in brief.

Antioxidant and Antisteatotic Activities of a New Fucoidan Extracted from Ferula hermonis Roots Harvested on Lebanese Mountains.

Fucoidan is a fucose-rich sulfated polysaccharide with attractive therapeutic potential due to a variety of biological activities, including antioxidant action. Fucoidan is typically found in the cell wall of marine brown algae, but extra-algal sources have also been discovered. In the present work, for the first time we extracted a water soluble fucoidan fraction from the roots of the terrestrial shrub Ferula hermonis. This fucoidan fraction was termed FUFe, and contained fucose, glucose, sulfate, smaller amounts of monosaccharides such as galactose and mannose, and a minor quantity of proteins. FUFe structural features were investigated by FTIR, 1H NMR and 13C NMR spectroscopy. The antioxidant property of FUFe was measured by DPPH, ABTS and FRAP assays, which revealed a high radical scavenging capacity that was confirmed in in vitro cellular models. In hepatic and endothelial cells, 50 μg/mL FUFe could reduce ROS production induced by intracellular lipid accumulation. Moreover, in hepatic cells FUFe exhibited a significant antisteatotic action, being able to reduce intracellular triglyceride content and to regulate the expression of key genes of hepatic lipid metabolism. Altogether, our results candidate FUFe as a possible bioactive compound against fatty liver disease and related vascular damage.

β-Glucan-Induced IL-10 Secretion by Monocytes Triggers Porcine NK Cell Cytotoxicity.

Beta-glucans are naturally occurring polysaccharides present in cell walls of fungi, yeast, bacteria, cereals, seaweed, and algae. These microbe-associated molecular patterns (MAMPs) possess immunomodulatory properties. In human, it has been suggested that NK cells can be activated by β-glucans. Here, we aimed to elucidate whether β-glucans modulate porcine NK cell responses in vitro and if so, how these effects are mediated. We investigated the effect of two β-glucans, Macrogard and Curdlan, which differ in solubility and structure. Direct addition of β-glucans to purified porcine NK cells did not affect cytotoxicity of these cells against K562 target cells. However, when using PBMC instead of purified NK cells, β-glucan addition significantly increased NK cell-mediated cytotoxicity. This effect depended on factors secreted by CD14+ monocytes upon β-glucan priming. Further analysis showed that monocytes secrete TNF-α, IL-6, and IL-10 upon β-glucan addition. Of these, IL-10 turned out to play a critical role in β-glucan-triggered NK cell cytotoxicity, since depletion of IL-10 completely abrogated the β-glucan-induced increase in cytotoxicity. Furthermore, addition of recombinant IL-10 to purified NK cells was sufficient to enhance cytotoxicity. In conclusion, we show that β-glucans trigger IL-10 secretion by porcine monocytes, which in turn leads to increased NK cell cytotoxicity, and thereby identify IL-10 as a potent stimulus of porcine NK cell cytotoxicity.

Structural basis for the exolytic activity of polysaccharide lyase family 6 alginate lyase BcAlyPL6 from human gut microbe Bacteroides clarus

Alginate is the structural polysaccharide of the cell wall of brown algae, which is an important carbon source for marine life. The depolymerization of alginate is dependent on alginate lyases. Recent studies showed that the alginate utilization ability had been obtained by human gut microbes. In contrast to the great number of studies on alginate lyases from marine/soil organisms, studies on alginate lyases from gut microbes are still limited. Here, the structure of a polysaccharide lyase family 6 (PL6) alginate lyase from human gut microbe Bacteroides clarus was solved by X-ray crystallography, which represents the cluster of two-domain PL6 alginate lyases from Bacteroidetes. Similar with the two-domain alginate lyase AlyGC originated from marine bacterium, both the N terminal domain (NTD) and C terminal domain (CTD) of BcAlyPL6 show right-handed parallel β-helix fold. However, unlike AlyGC, which forms a homodimer, BcAlyPL6 functions as a monomer. Biochemical analysis indicates that the substrate binding affinity is mainly contributed by the NTD while the CTD of BcAlyPL6 is involved in the formation of −1 subsite, which is essential for substrate turnover rate. Furthermore, CTD is involved in shaping a closed catalytic pocket, and deletion of it leads to increased activity towards highly polymerized substrate. Structure comparison of PL6 family alginate lyases implies that the linkers of two-domain alginate lyases might have evolutionary relationship with the N/C terminal extension of single-domain lyases.

Heavy metals removal from dumpsite leachate by algae and cyanobacteria

A living consortium of green algae (Ankistrodesmus) and cyanobacteria (Nostoc and Anabaena) was tested for the removal of heavy metal ions (Fe (II), Cu (II), Pb (II), Cr (VI) and Zn (II)) from dumpsite leachate under natural climatic conditions. Metal removal experiment was performed at 5 and 10 mg L−1 initial metal concentration in leachate and 0.8 and 1.6 g L−1 of biomass doses. Metal removal was in the order of Pb > Cu > Zn > Fe > Cr. Among all the metals algae and cyanobacteria removed the highest amount of Pb (7.03 ± 0.6 mgg−1) at 10 mgL−1 initial metal concentration and algal dose of 0.8 gL−1 and, the lowest amount of Cr metal (0.98 ± 0.6 mgg−1) was taken up at 5 mgL−1 initial metal concentration and 1.8 gL−1 of biomass dose. Metal removal by unit biomass mass (mgg−1) increases with an increase in initial metal concentration in leachate and decrease in biomass concentration however, the removal rates (percent removal per unit time) of metal ions has positive relationship with the initial biomass concentration. With an increase in initial metal concentration, the chances of collision between metal ions and available cations at the cell surface of biomass increases, resulting in increased metal uptake until the equilibrium is achieved after which the metal uptake remains constant. At small initial doses relatively a large number of adsorption sites at cell walls of algae and cyanobacteria are exposed, shifting the sorption equilibrium toward the biomass side resulting in rapid saturation of binding sites and increased bio sorption capacity of algae and cyanobacteria. This study reveals that living consortium of algae and cyanobacteria can be used as potential organisms for heavy metal removal from dumpsite leachate under natural climatic conditions.

Enzymatic hydrolysis of the industrial solid residue of red seaweed after agar extraction: Extracts characterization and modelling

The efficiency of enzymatic hydrolysis for the release of different biocompounds such as total phenolic compounds (TPC), sugars and proteins from the industrial solid residue of G. sesquipedale after agar extraction has been evaluated. Cellulase (0.25–8%, w/w, enzyme: solid residue, pH=5) has been proved to be an efficient enzyme to degrade the algae cell wall improving the release of the bound TPC with values up to 7.5mg gallic acid equivalent/g dry macroalgae residue. Monomer and oligomer carbohydrates were released to the reaction medium, namely glucose, galactose and arabinose with higher yields by increasing cellulase concentration. Enzyme combinations with other hydrolytic enzymes, such as xylanase and protease, did not bring any improvement of the TPC and sugar yields. Protein was also released to the enzymatic medium with protein extraction yields around 30%. The use of protease under basic conditions led to an increase in the release of the protein fraction and of the free amino acids content with a hydrophobic ratio higher than in the raw material. The kinetics of TPC and protein hydrolysis have been fitted to the power law and the Weibull models yielding the Weibull model the best fitting quality.

Antibacterial Properties of Fucoidans from the Brown Algae Fucus vesiculosus L. of the Barents Sea.

Simple SummaryThe continuous emergence of new pathogenic strains of bacteria resistant to antibiotics highlights the importance of search for the new natural sources of antimicrobial compounds. Fucoidans, sulfated polysaccharides from the cell walls of brown algae, attract the attention of many researchers due to their wide range of biological activities. Being natural and practically non-toxic, they are considered as a promising antimicrobial component that could, where possible, replace the use of strong chemical antiseptics and modern antibiotics. Natural fucoidans are polydisperse polysaccharides whose composition can vary greatly depending on the species, habitat, as well as on the way of purification. In this work, we investigate the structure and antibacterial properties of fucoidans from the brown algae Fucus vesiculosus gathered in the littoral of the Barents Sea. These fucoidans appear to have a significant bacteriostatic effect on the growth of four tested bacterial strains. Such an effect was more pronounced for crude fucoidan samples that did not undergo additional steps of purification.Fucoidans, sulfated polysaccharides found in cell walls of brown algae, are considered as a promising antimicrobial component for various applications in medicine and the food industry. In this study, we compare the antibacterial properties of two fractions of fucoidan from the brown algae Fucus vesiculosus gathered in the littoral of the Barents Sea and sampled at different stages of purification. The crude fraction of fucoidan was isolated from algae by extraction with aqueous ethanol and sonication. The purified fraction was obtained by additional treatment of the crude fraction with a solution of calcium chloride. The structural features of both fractions were characterized in detail and their antibacterial effects against several Gram-positive and Gram-negative bacteria were compared by photometry, acridine orange staining assay, and atomic force microscopy. Fucoidan inhibited growth in all of the above microorganisms, showing a bacteriostatic effect with minimum inhibitory concentrations (MIC) in the range between 4 and 6 mg/mL, with E. coli being the most sensitive to both fractions. Changes in the chemical composition after treatment of the crude fraction with a solution of calcium chloride led to a decrease in the content of sulfates and uronic acids and diminished antibacterial activity.

Mechanism of nanotoxicity in Chlorella vulgaris exposed to zinc and iron oxide.

Usage of nanoparticle in various products has increased tremendously in the recent past. Toxicity of these nanoparticles can have a huge impact on aquatic ecosystem. Algae are the ideal organism of the aquatic ecosystem to understand the toxicity impact of nanoparticles. The present study focuses on the toxicity evaluation of zinc oxide (ZnO) and iron oxide (Fe2O3) nanoparticles towards freshwater microalgae, Chlorella vulgaris. The dose dependent growth retardation in Chlorella vulgaris is observed under ZnO and Fe2O3 nanoparticles and nanoform attributed more toxicity than their bulk counterparts. The IC50 values of ZnO and Fe2O3 nanoparticles was reported at 0.258 mg L-1 and 12.99 mg L-1 whereas, for the bulk-form, it was 1.255 mgL-1 and 17.88 mg L-1, respectively. The significant decline in chlorophyll content and increase in proline content, activity of superoxide dismutase and catalase, indicated the stressful physiological state of microalgae. An increased lactate dehydrogenase level in treated samples suggested membrane disintegration by ZnO and Fe2O3 nanoparticles. Compound microscopy, scanning electron microscopy and transmission electron microscopy confirm cell entrapment, deposition of nanoparticles on the cell surface and disintegration of algal cell wall. Higher toxicity of nanoform in comparison to bulk chemistry is a point of concern.

2020 FASEB Science Research Conference on Microbial Glycobiology, July 13-14, 2020.

2020 FASEB Science Research Conference on Microbial Glycobiology, July 13-14, 2020.

Diversity of Sulfated Polysaccharides From Cell Walls of Coenocytic Green Algae and Their Structural Relationships in View of Green Algal Evolution.

Seaweeds biosynthesize sulfated polysaccharides as key components of their cell walls. These polysaccharides are potentially interesting as biologically active compounds. Green macroalgae of the class Ulvophyceae comprise sulfated polysaccharides with great structural differences regarding the monosaccharide constituents, linearity of their backbones, and presence of other acidic substituents in their structure, including uronic acid residues and pyruvic acid. These structures have been thoroughly studied in the Ulvales and Ulotrichales, but only more recently have they been investigated with some detail in ulvophytes with giant multinucleate (coenocytic) cells, including the siphonous Bryopsidales and Dasycladales, and the siphonocladous Cladophorales. An early classification of these structurally heterogeneous polysaccharides was based on the presence of uronic acid residues in these molecules. In agreement with this classification based on chemical structures, sulfated polysaccharides of the orders Bryopsidales and Cladophorales fall in the same group, in which this acidic component is absent, or only present in very low quantities. The cell walls of Dasycladales have been less studied, and it remains unclear if they comprise sulfated polysaccharides of both types. Although in the Bryopsidales and Cladophorales the most important sulfated polysaccharides are arabinans and galactans (or arabinogalactans), their major structures are very different. The Bryopsidales produce sulfated pyruvylated 3-linked β-d-galactans, in most cases, with ramifications on C6. For some species, linear sulfated pyranosic β-l-arabinans have been described. In the Cladophorales, also sulfated pyranosic β-l-arabinans have been found, but 4-linked and highly substituted with side chains. These differences are consistent with recent molecular phylogenetic analyses, which indicate that the Bryopsidales and Cladophorales are distantly related. In addition, some of the Bryopsidales also biosynthesize other sulfated polysaccharides, i.e., sulfated mannans and sulfated rhamnans. The presence of sulfate groups as a distinctive characteristic of these biopolymers has been related to their adaptation to the marine environment. However, it has been shown that some freshwater algae from the Cladophorales also produce sulfated polysaccharides. In this review, structures of sulfated polysaccharides from bryopsidalean, dasycladalean, and cladophoralean green algae studied until now are described and analyzed based on current phylogenetic understanding, with the aim of unveiling the important knowledge gaps that still exist.

Therapeutic Uses of Red Macroalgae.

Red Seaweed “Rhodophyta” are an important group of macroalgae that include approximately 7000 species. They are a rich source of structurally diverse bioactive constituents, including protein, sulfated polysaccharides, pigments, polyunsaturated fatty acids, vitamins, minerals, and phenolic compounds with nutritional, medical, and industrial importance. Polysaccharides are the main components in the cell wall of red algae and represent about 40–50% of the dry weight, which are extensively utilized in industry and pharmaceutical compounds, due to their thickening and gelling properties. The hydrocolloids galactans carrageenans and agars are the main red seaweed cell wall polysaccharides, which had broad-spectrum therapeutic characters. Generally, the chemical contents of seaweed are different according to the algal species, growth stage, environment, and external conditions, e.g., the temperature of the water, light intensity, nutrient concentrations in the ecosystem. Economically, they can be recommended as a substitute source for natural ingredients that contribute to a broad range of bioactivities like cancer therapy, anti-inflammatory agents, and acetylcholinesterase inhibitory. This review touches on the main points of the pharmaceutical applications of red seaweed, as well as the exploitation of their specific compounds and secondary metabolites with vital roles.

Antiviral efficacy of orally delivered neoagarohexaose, a nonconventional TLR4 agonist, against norovirus infection in mice

The neoagarohexaose (NA6) is an oligosaccharide that is derived from agarose, the major component of red algae cell walls, by enzymatic hydrolysis. Here we show that NA6 is a noncanonical Toll-like receptor 4 (TLR4) agonist with antiviral activity against norovirus. Its TLR4 activation was dependent on myeloid differentiation factor 2 (MD2) and cluster of differentiation 14 (CD14), leading to interferon-β (IFN-β) and tumor necrosis factor-α (TNF-α) production. This effect was abolished by TLR4 knockdown or knockout in murine macrophages. NA6 inhibited murine norovirus (MNV) replication with an EC50 of 1.5 μM in RAW264.7 cells. It also lowered viral RNA titer in a human hepatocellular carcinoma Huh7-derived cell line harboring a human norovirus subgenomic replicon. The antiviral activity of NA6 was mainly attributed to IFN-β produced through the TLR4-TRIF signaling pathway. NA6-induced TNF-α, which had little effect on norovirus replication per se, primed macrophages to mount greater antiviral innate immune responses when IFN signaling was activated. NA6 boosted the induction of IFN-β in MNV-infected RAW264.7 cells and upregulated IFN-regulatory factor-1, an IFN-stimulated gene. NA6 induced IFN-β expression in the distal ileum with Peyer's patches and oral administration of NA6 reduced MNV loads through activation of TLR4 signaling, highlighting its potential contribution to protective antiviral innate immunity against norovirus.

Preferred crystallographic orientation of cellulose in plant primary cell walls

Cellulose, the most abundant biopolymer on earth, is a versatile, energy rich material found in the cell walls of plants, bacteria, algae, and tunicates. It is well established that cellulose is crystalline, although the orientational order of cellulose crystallites normal to the plane of the cell wall has not been characterized. A preferred orientational alignment of cellulose crystals could be an important determinant of the mechanical properties of the cell wall and of cellulose-cellulose and cellulose-matrix interactions. Here, the crystalline structures of cellulose in primary cell walls of onion (Allium cepa), the model eudicot Arabidopsis (Arabidopsis thaliana), and moss (Physcomitrella patens) were examined through grazing incidence wide angle X-ray scattering (GIWAXS). We find that GIWAXS can decouple diffraction from cellulose and epicuticular wax crystals in cell walls. Pole figures constructed from a combination of GIWAXS and X-ray rocking scans reveal that cellulose crystals have a preferred crystallographic orientation with the (200) and (110)/(1bar 10) planes preferentially stacked parallel to the cell wall. This orientational ordering of cellulose crystals, termed texturing in materials science, represents a previously unreported measure of cellulose organization and contradicts the predominant hypothesis of twisting of microfibrils in plant primary cell walls.

Characterization of sponge-associated Verrucomicrobia: microcompartment-based sugar utilization and enhanced toxin-antitoxin modules as features of host-associated Opitutales.

Bacteria of the phylum Verrucomicrobia are ubiquitous in marine environments and can be found as free-living organisms or as symbionts of eukaryotic hosts. Little is known about host-associated Verrucomicrobia in the marine environment. Here we reconstructed two genomes of symbiotic Verrucomicrobia from bacterial metagenomes derived from the Atlanto-Mediterranean sponge Petrosia ficiformis and three genomes from strains that we isolated from offshore seawater of the Eastern Mediterranean Sea. Phylogenomic analysis of these five strains indicated that they are all members of Verrucomicrobia subdivision 4, order Opitutales. We compared these novel sponge-associated and seawater-isolated genomes to closely related Verrucomicrobia. Genomic analysis revealed that Planctomycetes-Verrucomicrobia microcompartment gene clusters are enriched in the genomes of symbiotic Opitutales including sponge symbionts but not in free-living ones. We hypothesize that in sponge symbionts these microcompartments are used for degradation of l-fucose and l-rhamnose, which are components of algal and bacterial cell walls and therefore may be found at high concentrations in the sponge tissue. Furthermore, we observed an enrichment of toxin-antitoxin modules in symbiotic Opitutales. We suggest that, in sponges, verrucomicrobial symbionts utilize these modules as a defence mechanism against antimicrobial activity deriving from the abundant microbial community co-inhabiting the host.