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Today, understanding of cell wall evolution is incomplete due to limited knowledge of cell wall structure of non-flowering plants. Besides polysaccharides, proteoglycans like arabinogalactan-proteins are important constituents of the cell wall of most if not all seed plants. This article provides the first detailed characterization of AGPs from a lycophyte and different ferns, which are the closest living relatives to seed plants. The amount of protein in fern AGPs (6–12%) was in a range comparable to most seed plant AGPs. However, it was higher for Lycopodium AGP (17%). The carbohydrate moieties of investigated AGPs showed typical features known for type-II arabinogalactans from seed plants and showed cross-reaction with polyclonal antibodies raised against Echinacea AGP. On the other hand, unusual structural components like high quantities of pyranosidic arabinose residues for Lycopodium AGP and 3- O -methyl-rhamnose for fern AGPs have been detected. [ABSTRACT FROM AUTHOR]

Abstract: This investigation shows structural features of two macromolecules from roots of Echinacea pallida (Nutt.) Nutt: an arabinogalactan-protein (AGP) and an arabinan. The arabinogalactan-protein was precipitated with β-glucosyl Yariv reagent from a high molecular weight fraction. Investigations of the neutral sugar composition revealed Gal (52.1% w/w) and Ara (38.2% w/w) in a ratio of 1.4:1, accompanied by Glc (6.9% w/w) and Rha (2.8% w/w). The content of uronic acids was 6.2%. Mild acid hydrolysis detects Ara and Glc being located at the periphery of the molecule. Linkage analyses and NMR spectroscopy revealed a backbone of the polysaccharide mainly consisting of 3-linked and 3,6-linked Galp-residues. Side chains are composed of 3,6-linked or 6-linked Galp terminating in 5-linked Araf, terminal Araf, Glcp and GlcAp. The protein part (3.9% w/w) of the AGP is rich in Hyp, Ser, Ala, Thr, Glu, Asp and Gly. The amount of Hyp was determined by a colorimetric method and found to be (0.65% (w/w) of the AGP, which is in good agreement with the result obtained by amino acid hydrolysis (0.67% w/w). The arabinan was isolated from the supernatant of the Yariv precipitation on the basis of solubility in EtOH (80%). It mainly consists of Ara (85.8%). Linkage analyses and NMR spectroscopy indicate a highly branched molecule, consisting of 3,5-linked, 5-linked and terminal Araf-residues in equal amounts. [Copyright &y& Elsevier]

An important step for plant diversification was the transition from freshwater to terrestrial habitats. The bryophytes and all vascular plants share a common ancestor that was probably the first to adapt to life on land. A polysaccharide-rich cell wall was necessary to cope with newly faced environmental conditions. Therefore, some pre-requisites for terrestrial life have to be shared in the lineages of modern bryophytes and vascular plants. This review focuses on hornwort and liverwort cell walls and aims to provide an overview on shared and divergent polysaccharide features between these two groups of bryophytes and vascular plants. Analytical, immunocytochemical, and bioinformatic data were analysed. The major classes of polysaccharides—cellulose, hemicelluloses, and pectins—seem to be present but have diversified structurally during evolution. Some polysaccharide groups show structural characteristics which separate hornworts from the other bryophytes or are too poorly studied in detail to be able to draw absolute conclusions. Hydroxyproline-rich glycoprotein backbones are found in hornworts and liverworts, and show differences in, for example, the occurrence of glycosylphosphatidylinositol (GPI)-anchored arabinogalactan-proteins, while glycosylation is practically unstudied. Overall, the data are an appeal to researchers in the field to gain more knowledge on cell wall structures in order to understand the changes with regard to bryophyte evolution. [ABSTRACT FROM AUTHOR]

Streptophyte algae are the closest relatives to land plants; their latest common ancestor performed the most drastic adaptation in plant evolution around 500 million years ago: the conquest of land. Besides other adaptations, this step required changes in cell wall composition. Current knowledge on the cell walls of streptophyte algae and especially on the presence of arabinogalactan‐proteins (AGPs), important signalling molecules in all land plants, is limited. To get deeper insights into the cell walls of streptophyte algae, especially in Charophyceae, we performed sequential cell wall extractions of four Chara species. The three species Chara globularis, Chara subspinosa and Chara tomentosa revealed comparable cell wall compositions, with pectins, xylans and xyloglucans, whereas Chara aspera stood out with higher amounts of uronic acids in the pectic fractions and lack of reactivity with antibodies binding to xylan‐ and xyloglucan epitopes. Search for AGPs in the four Chara species and in Nitellopsis obtusa revealed the presence of galactans with pyranosidic galactose in 1,3‐, 1,6‐ and 1,3,6‐linkage, which are typical galactan motifs in land plant AGPs. A unique feature of these branched galactans was high portions of 3‐O‐methylgalactose. Only Nitellopsis contained substantial amounts of arabinose A bioinformatic search for prolyl‐4‐hydroxylases, involved in the biosynthesis of AGPs, revealed one possible functional sequence in the genome of Chara braunii, but no hydroxyproline could be detected in the four Chara species or in Nitellopsis obtusa. We conclude that AGPs that is typical for land plants are absent, at least in these members of the Charophyceae. [ABSTRACT FROM AUTHOR]

SUMMARY: Significant changes have occurred in plant cell wall composition during evolution and diversification of tracheophytes. As the sister lineage to seed plants, knowledge on the cell wall of ferns is key to track evolutionary changes across tracheophytes and to understand seed plant‐specific evolutionary innovations. Fern cell wall composition is not fully understood, including limited knowledge of glycoproteins such as the fern arabinogalactan proteins (AGPs). Here, we characterize the AGPs from the leptosporangiate fern genera Azolla, Salvinia, and Ceratopteris. The carbohydrate moiety of seed plant AGPs consists of a galactan backbone including mainly 1,3‐ and 1,3,6‐linked pyranosidic galactose, which is conserved across the investigated fern AGPs. Yet, unlike AGPs of angiosperms, those of ferns contained the unusual sugar 3‐O‐methylrhamnose. Besides terminal furanosidic arabinose, Ara (Araf), the main linkage type of Araf in the ferns was 1,2‐linked Araf, whereas in seed plants 1,5‐linked Araf is often dominating. Antibodies directed against carbohydrate epitopes of AGPs supported the structural differences between AGPs of ferns and seed plants. Comparison of AGP linkage types across the streptophyte lineage showed that angiosperms have rather conserved monosaccharide linkage types; by contrast bryophytes, ferns, and gymnosperms showed more variability. Phylogenetic analyses of glycosyltransferases involved in AGP biosynthesis and bioinformatic search for AGP protein backbones revealed a versatile genetic toolkit for AGP complexity in ferns. Our data reveal important differences across AGP diversity of which the functional significance is unknown. This diversity sheds light on the evolution of the hallmark feature of tracheophytes: their elaborate cell walls. [ABSTRACT FROM AUTHOR]

Background: The polyphyletic group of seagrasses shows an evolutionary history from early monocotyledonous land plants to the marine environment. Seagrasses form important coastal ecosystems worldwide and large amounts of seagrass detritus washed on beaches might also be valuable bioeconomical resources. Despite this importance and potential, little is known about adaptation of these angiosperms to the marine environment and their cell walls. Results: We investigated polysaccharide composition of nine seagrass species from the Mediterranean, Red Sea and eastern Indian Ocean. Sequential extraction revealed a similar seagrass cell wall polysaccharide composition to terrestrial angiosperms: arabinogalactans, pectins and different hemicelluloses, especially xylans and/or xyloglucans. However, the pectic fractions were characterized by the monosaccharide apiose, suggesting unusual apiogalacturonans are a common feature of seagrass cell walls. Detailed analyses of four representative species identified differences between organs and species in their constituent monosaccharide composition and lignin content and structure. Rhizomes were richer in glucosyl units compared to leaves and roots. Enhalus had high apiosyl and arabinosyl abundance, while two Australian species of Amphibolis and Posidonia, were characterized by high amounts of xylosyl residues. Interestingly, the latter two species contained appreciable amounts of lignin, especially in roots and rhizomes whereas Zostera and Enhalus were lignin-free. Lignin structure in Amphibolis was characterized by a higher syringyl content compared to that of Posidonia. Conclusions: Our investigations give a first comprehensive overview on cell wall composition across seagrass families, which will help understanding adaptation to a marine environment in the evolutionary context and evaluating the potential of seagrass in biorefinery incentives. [ABSTRACT FROM AUTHOR]

SUMMARY: Charophyte green algae (CGA) are assigned to be the closest relatives of land plants and therefore enlighten processes in the colonization of terrestrial habitats. For the transition from water to land, plants needed significant physiological and structural changes, as well as with regard to cell wall composition. Sequential extraction of cell walls of Nitellopsis obtusa (Charophyceae) and Spirogyra pratensis (Zygnematophyceae) offered a comparative overview on cell wall composition of late branching CGA. Because arabinogalactan‐proteins (AGPs) are considered common for all land plant cell walls, we were interested in whether these special glycoproteins are present in CGA. Therefore, we investigated both species with regard to characteristic features of AGPs. In the cell wall of Nitellopsis, no hydroxyproline was present and no AGP was precipitable with the β‐glucosyl Yariv's reagent (βGlcY). By contrast, βGlcY precipitation of the water‐soluble cell wall fraction of Spirogyra yielded a glycoprotein fraction rich in hydroxyproline, indicating the presence of AGPs. Putative AGPs in the cell walls of non‐conjugating Spirogyra filaments, especially in the area of transverse walls, were detected by staining with βGlcY. Labelling increased strongly in generative growth stages, especially during zygospore development. Investigations of the fine structure of the glycan part of βGlcY‐precipitated molecules revealed that the galactan backbone resembled that of AGPs with 1,3‐ 1,6‐ and 1,3,6‐linked Galp moieties. Araf was present only in small amounts and the terminating sugars consisted predominantly of pyranosidic terminal and 1,3‐linked rhamnose residues. We introduce the term 'rhamnogalactan‐protein' for this special AGP‐modification present in S. pratensis. [ABSTRACT FROM AUTHOR]

Abstract: Mucilage is thought to play a role in salinity tolerance in certain halophytic species by regulating water ascent and ion transport. The localization and composition of mucilage in the halophyte Kosteletzkya virginica was therefore investigated. Plants were grown in a hydroponic system in the presence or absence of 100mM NaCl and regularly harvested for growth parameter assessment and mucilage analysis with the gas liquid chromatography method. NaCl treatment stimulated shoot growth and biomass accumulation, had little effect on shoot and root water content, and reduced leaf water potential (Ψ w), osmotic potential (Ψ s) as well as stomatal conductance (g s). Mucilage increased in shoot, stems and roots in response to salt stress. Furthermore, changes were also observed in neutral monosaccharide components. Levels of rhamnose and uronic acid increased with salinity. Staining with a 0.5% alcian blue solution revealed the presence of mucopolyssacharides in xylem vessels and salt-induced mucilaginous precipitates on the leaf abaxial surface. Determination of ion concentrations showed that a significant increase of Na+ and a decrease of K+ and Ca2+ simultaneously occurred in tissues and in mucilage under salt stress. Considering the high proportion of rhamnose and uronic acid in stem mucilage, we suggest that the pectic polysaccharide could be involved in Na+ fixation, though only a minor fraction of accumulated sodium appeared to be firmly bound to mucilage. [Copyright &y& Elsevier]

• AGPs from gymnosperms Ginkgo, Cycas, Ephedra and Encephalartos were characterized. • The unusual monosaccharide acofriose was found in AGPs from Ginkgo, Ephedra and Encephalartos. • A unique attribute of Ginkgo AGP is a glycan part mixed of type I and II AGs. • Bioinformatic investigations on glycosyltransferases complete analytical results. • The study contributes to understanding of evolution of AGPs during life on land. Arabinogalactan-proteins (AGPs), important signalling molecules of the plant cell wall, are structurally extensively investigated in angiosperms, but information on AGPs in gymnosperms is still limited. We characterized AGPs from the gymnosperms Ginkgo biloba , Ephedra distachya , Encephalartos longifolius and Cycas revoluta. The protein contents are comparable to that of angiosperm AGPs. Hydroxyproline is the site of linking the carbohydrate part and was detected in all AGPs with highest concentration in Cycas AGP (1.1 % of the AGP). Interestingly, with the exception of Cycas , all AGPs contained the monosaccharide 3- O -methylrhamnose not present in angiosperm polysaccharides. The carbohydrate moieties of Cycas and Ephredra showed the main components 1,3,6-linked galactose and terminal arabinose typical of angiosperm AGPs, whereas that of Ginkgo AGP was unique with 1,4-linked galactose as dominant structural element. Bioinformatic search for glycosyltransferases in Ginkgo genome also revealed a lower number of galactosyltransferases responsible for biosynthesis of the 1,3-Gal/1,6-Gal AGP backbone. [ABSTRACT FROM AUTHOR]

Seagrasses evolved from monocotyledonous land plants that returned to the marine habitat. This transition was accomplished by substantial changes in cell wall composition, revealing habitat-driven adaption to the new environment. Whether arabinogalactan-proteins (AGPs), important signalling molecules of land plants, are present in seagrass cell walls is of evolutionary and plant development interest. AGPs of Zostera marina L. were isolated and structurally characterised by analytical and bioinformatics methods as well as by ELISA with different anti-AGP antibodies. Calcium-binding capacity of AGPs was studied by isothermal titration calorimetry (ITC) and microscopy. Bioinformatic searches of the Z. marina proteome identified 9 classical AGPs and a large number of chimeric AGPs. The glycan structures exhibit unique features, including a high degree of branching and an unusually high content of terminating 4-O-methyl-glucuronic acid (4-OMe GlcA) residues. Although the common backbone structure of land plant AGPs is conserved in Z. marina, the terminating residues are distinct with high amounts of uronic acids. These differences likely result from the glycan-active enzymes (glycosyltransferases and methyltransferases) and are essential for calcium-binding properties. The role of this polyanionic surface is discussed with regard to adaption to the marine environment. [ABSTRACT FROM AUTHOR]

Arabinogalactan-proteins (AGPs) are important proteoglycans of plant cell walls. They seem to be present in most, if not all seed plants, but their occurrence and structure in bryophytes is widely unknown and actually the focus of AGP research. With regard to evolution of plant cell wall, we isolated AGPs from the three mosses Sphagnum sp., Physcomitrella patens and Polytrichastrum formosum . The moss AGPs show structural characteristics common for AGPs of seed plants, but also unique features, especially 3- O -methyl-rhamnose (trivial name acofriose) as terminal monosaccharide not found in arabinogalactan-proteins of angiosperms and 1,2,3-linked galactose as branching point never found in arabinogalactan-proteins before. [ABSTRACT FROM AUTHOR]

Root extracts of the medicinal plant Pelargonium sidoides , native to South Africa, are used globally for the treatment of common cold and cough. Due to an increasing economic commercialization of P. sidoides remedies, wild collections of root material should be accompanied by effective methods for plant propagation like somatic embryogenesis. Based on this, the influence of arabinogalactan-proteins (AGPs) on somatic embryogenesis and plant propagation of P. sidoides has been investigated. High-molecular weight AGPs have been isolated from dried roots as well as from cell cultures of P. sidoides with yields between 0.1% and 0.9%, respectively. AGPs are characterized by a 1,3-linked Gal p backbone, branched at C6 to 1,6-linked Gal p side chains terminated by Ara f and to a minor extent by Glc p A, Gal p or Rha p . Treatment of explants of P. sidoides with AGPs from roots or suspension culture over 5.5 weeks resulted in effective stimulation of somatic embryo development and plant regeneration. [ABSTRACT FROM AUTHOR]

Arabinogalactan (AG) isolated from dust of a traditional farm prevents disease in murine models of allergy. However, it is unclear whether this polysaccharide has immune regulatory properties in humans. The aim of this study w as to test the influence of AG on the immune-stimulating properties of human dendritic cells (DCs). Moreover, we sought to identify the receptor to which AG binds. AG was produced from plant callus tissue under sterile conditions to avoid the influence of pathogen-associated molecular patterns in subsequent experiments. The influence of AG on the human immune system was investigated by analyzing its impact on monocyte-derived DCs. To analyze whether the T cell stimulatory capacity of AG-stimulated DCs is altered, an MLR with naive Th cells was performed. We revealed that AG reduced T cell proliferation in a human MLR. In the search for a molecular mechanism, we found that AG binds to the immune modulatory receptors DC-specific ICAM-3-grabbing non integrin (DC-SIGN) and macrophage mannose receptor 1 (MMR-1). Stimulation of these receptors with AG simultaneously with TLR4 stimulation with LPS increased the expression of the E3 ubiquitin-protein ligase tripartite motif-containing protein 21 and decreased the phosphorylation of NF-κB p65 in DCs. This led to a reduced activation profile with reduced costimulatory molecules and proinflammatory cytokine production. Blocking of MMR-1 or DC-SIGN with neutralizing Abs partially inhibits this effect. We conclude that AG dampens the activation of human DCs by LPS via binding to DC-SIGN and MMR-1, leading to attenuated TLR signaling. This results in a reduced T cell activation capacity of DCs. [ABSTRACT FROM AUTHOR]

Abstract: An arabinogalactan from wood of the larch tree (Larix laricina), which is a source of dietary fiber, is composed of d-galactose and l-arabinose in a 6:1 molar ratio accompanied by small amounts of d-glucuronic acid. The molecular mass has been determined to be 38kDa. Linkage analysis in combination with partial hydrolysis and spectroscopic investigations revealed a 1,3-linked Galp backbone, branched at C6 to 1,6-linked Galp side residues terminated by Galp, Arap, Araf or GlcpA. Monoclonal antibodies directed against a type-II-arabinogalactan from Echinacea purpurea showed cross reactivity with the larch arabinogalactan and have been used to develop an ELISA for quantification of this polysaccharide. [Copyright &y& Elsevier]

Abstract: An arabinogalactan-protein (AGP) from whole grain of oat (Avena sativa L.) has been isolated for the first time by double precipitation with β-glucosyl Yariv reagent and characterized with regard to its polysaccharide and protein part. The large carbohydrate moiety is rich in galactose (63.0% w/w) and arabinose (32.8% w/w) and free of uronic acids. Linkage analysis of AGP and its products obtained by partial acid hydrolysis as well as enzymatic degradation with specific recombinant enzymes revealed that the carbohydrate moiety consists of a 1,3-Galp backbone and is linked in position 6 to short 1,6-Galp chains, terminating in Araf. In the protein part, high contents of alanine, hydroxyproline and serine could be found which are typical for AGPs. The molecular mass of AGP was determined to be 83 kDa. The carbohydrate moieties, released by alkaline degradation of the protein part, had a size of about 7 kDa. Consequently, the overall structure of the AGP from oat could be assigned to be consistent with the wattle-blossom model of AGPs. [Copyright &y& Elsevier]

Galectins represent β-galactoside-binding proteins with numerous functions. Due to their role in tumor progression, human galectins-1, -3 and -7 (Gal-1, -3 and -7) are potential targets for cancer therapy. As plant derived glycans might act as galectin inhibitors, we prepared galactans by partial degradation of plant arabinogalactan-proteins. Besides commercially purchased galectins, we produced Gal-1 and -7 in a cell free system and tested binding capacities of the galectins to the galactans by biolayer-interferometry. Results for commercial and cell-free expressed galectins were comparable confirming functionality of the cell-free produced galectins. Our results revealed that galactans from Echinacea purpurea bind to Gal-1 and -7 with KD values of 1–2 µM and to Gal-3 slightly stronger with KD values between 0.36 and 0.70 µM depending on the sensor type. Galactans from the seagrass Zostera marina with higher branching of the galactan and higher content of uronic acids showed stronger binding to Gal-3 (0.08–0.28 µM) compared to galactan from Echinacea. The results contribute to knowledge on interactions between plant polysaccharides and galectins. Arabinogalactan-proteins have been identified as a new source for production of galactans with possible capability to act as galectin inhibitors. [ABSTRACT FROM AUTHOR]

This study intends to investigate the inhibitory potential of different plant derived saccharides on cell migration and adhesion of pancreatic ductal adenocarcinoma (PDAC) cells to microvascular liver endothelium, particularly considering the role of transmembranous galectin-3. PDAC cell lines PancTu1 and Panc1 were characterized by considerable (transmembranous) galectin-3 (Gal3) expression. SiRNA mediated Gal3 knockdown as well as treatment with differentially processed pectins and arabinogalactan-proteins (AGPs) did not impact on cell migration of either PDAC cell line. In contrast, Gal3 knockdown reduced adhesion of PDAC cells to the liver endothelial cell line TMNK-1 being more pronounced in Panc1 cells. Similarly, plant derived substances did not impact cell adhesion of PancTu1 cells while partially hydrolyzed citrus pectin (MCP), pectinase-treated MCP (MCP Pec) and partially hydrolized AGP (AGP TFA) clearly diminished adhesive properties of Panc1 cells. MCP Pec or AGP TFA could not further intensify the adhesion reducing effect of galectin-3 knockdown, indicating that these plant derived polysaccharides are able to inhibit PDAC cell adhesion to liver endothelial cells in a galectin-3 dependent manner. Overall, these data suggest an inhibitory potential of plant derived processed saccharides which have undergone chemical modification in impairing PDAC cell adhesion to liver endothelium. Image 1 • An in vitro testsystem for adhesion of pancreatic ductal adenocarcinoma (PDAC) to liver cells has been developed. • The role of transmembranous galectin-3 on this process is in the focus of this work. • SiRNA mediated Gal3 knockdown reduced adhesion of PDAC cells to the liver cell line TMNK-1. • Modified plant galactans clearly diminished adhesive properties of PDAC cells. • Overall, these data suggest an inhibitory potential of plant derived processed galactans on metastasis. [ABSTRACT FROM AUTHOR]