Neochloris oleoabundans cell walls have an altered composition when cultivated under different growing conditions

The impact that environmental factors have on the intracellular components of microalgae has been the focus of research for a number of decades. Despite that, their effects on the cell wall have received very little attention. In this study, we investigated how different growing conditions affect the cell walls of N. oleoabundans. The results revealed that the cell wall composition varied in that the modifications were different in the four cultivation media: freshwater nitrogen-replete (optimum culture) and -depleted conditions, and seawater nitrogen-replete and -depleted conditions. Nitrogen deficiency in freshwater cultivation was the only condition that significantly (p <.05) increased the total content of carbohydrates in the cell wall. The three most abundant components of freshwater-cultivated cell wall polysaccharides were rhamnose, galactose and glucuronic acid whereas in seawater media the main components of cell wall polysaccharides were rhamnose, glucose and galactose. The combined results of the biochemical analyses and monoclonal antibodies epitope-binding revealed that N. oleoabundans cell walls are likely composed of sulphated polysaccharides enriched in mannose, β-(1 → 4)-D-mannans, and glucose as they grow in seawater. Salinity and nitrogen deficiency also had an impact on the nitrogenous components of the cell wall. Under these conditions we observed a decrease in glucosamine in the cell wall. The analysis of specific binding of monoclonal antibodies, revealed that the cell wall of N. oleoabundans is possibly enriched in arabinogalactan proteins (AGPs). Under salinity and nitrogen deficiency N. oleoabundans increased the proportion of the non-polar to polar amino acids in the cell walls. An increase of leucine in the cell walls may suggest that N. oleoabundans contains leucine-rich repeat proteins which are known to play a vital role in stress responses. This report provides new insights into microalgae cell wall biology and how cell walls are remodelled when growing under different conditions.</p

Knowledge of learning disabilities: the relationship with choice, duty of care and non-aversive approaches

The present study examines the relationship between the knowledge of the diagnostic criteria for a learning disability (based on DSM IV criteria), care practices and experience in health care and social care staff. Responses to a questionnaire were analysed in terms of participants emphasis on: recognizing duty of care; enabling choice; non-aversive and aversive strategies. Results indicated that the knowledge of the criteria for a learning disability was limited, with only I6% of the sample correctly identifying all three criteria. There were no significant differences between the two groups in relation to experience or level of knowledge. No clear cut differences were found between the groups in relation to tendency to emphasize a particular management approach, with the strategies adopted appearing to be influenced by vignettes used in this study. Participants tended to give responses that identified both a recognition of their duty of care to clients and the need to enable choice. Limitations of this study are discussed

Monoclonal antibodies indicate low-abundance links between heteroxylan and other glycans of plant cell walls.

The derivation of two sensitive monoclonal antibodies directed to heteroxylan cell wall polysaccharide preparations has allowed the identification of potential inter-linkages between xylan and pectin in potato tuber cell walls and also between xylan and arabinogalactan-proteins in oat grain cell walls. Plant cell walls are complex composites of structurally distinct glycans that are poorly understood in terms of both in muro inter-linkages and developmental functions. Monoclonal antibodies (MAbs) are versatile tools that can detect cell wall glycans with high sensitivity through the specific recognition of oligosaccharide structures. The isolation of two novel MAbs, LM27 and LM28, directed to heteroxylan, subsequent to immunisation with a potato cell wall fraction enriched in rhamnogalacturonan-I (RG-I) oligosaccharides, is described. LM27 binds strongly to heteroxylan preparations from grass cell walls and LM28 binds to a glucuronosyl-containing epitope widely present in heteroxylans. Evidence is presented suggesting that in potato tuber cell walls, some glucuronoxylan may be linked to pectic macromolecules. Evidence is also presented that suggests in oat spelt xylan both the LM27 and LM28 epitopes are linked to arabinogalactan-proteins as tracked by the LM2 arabinogalactan-protein epitope. This work extends knowledge of the potential occurrence of inter-glycan links within plant cell walls and describes molecular tools for the further analysis of such links.This work was supported by the European Union Seventh Framework Programme (FP7 2007-2013) under the WallTraC project (Grant Agreement number 263916). (This article reflects the authors’ views only and the European Union is not liable for any use that may be made of the information contained herein). The work was also supported by the United Kingdom Biotechnology and Biological Research Council (BBSRC, Grant BB/K017489/1). JX acknowledges support from the Chinese Scholarship Council, TAT from a BBSRC studentship and MGR from the Danish Strategic Research Council and The Danish Council for Independent Research, Technology and Production Sciences as part of the GlycAct project (FI 10-093465). We acknowledge kind gifts of enzymes from Harry Gilbert and oligosaccharides from Sanna Koutaniemi. We thank Theodora Tryfona for mass spectrometry analysis.This is the final version of the article. It first appeared from Springer via http://dx.doi.org/10.1007/s00425-015-2375-

Synthesis of branched and linear 1,4-linked galactan oligosaccharides

Strategic naphthylidine protection allows for the rapid assembly of linear and branched 1,4-galactans.</p

Report on the Current Inventory of the Toolbox for Plant Cell Wall Analysis: Proteinaceous and Small Molecular Probes

Plant cell walls are highly complex structures composed of diverse classes of polysaccharides, proteoglycans, and polyphenolics, which have numerous roles throughout the life of a plant. Significant research efforts aim to understand the biology of this cellular organelle and to facilitate cell-wall-based industrial applications. To accomplish this, researchers need to be provided with a variety of sensitive and specific detection methods for separate cell wall components, and their various molecular characteristics in vitro as well as in situ. Cell wall component-directed molecular detection probes (in short: cell wall probes, CWPs) are an essential asset to the plant glycobiology toolbox. To date, a relatively large set of CWPs has been produced—mainly consisting of monoclonal antibodies, carbohydrate-binding modules, synthetic antibodies produced by phage display, and small molecular probes. In this review, we summarize the state-of-the-art knowledge about these CWPs; their classification and their advantages and disadvantages in different applications. In particular, we elaborate on the recent advances in non-conventional approaches to the generation of novel CWPs, and identify the remaining gaps in terms of target recognition. This report also highlights the addition of new “compartments” to the probing toolbox, which is filled with novel chemical biology tools, such as metabolic labeling reagents and oligosaccharide conjugates. In the end, we also forecast future developments in this dynamic field

A biology-driven approach for producing monoclonal antibodies against green algal cell wall components

A biology-driven approach for producing monoclonal antibodies against green algal cell wall components. 14th cell wall meetin

Production and fine characterization of new antibodies against rhamnogalacturonan I

Production and fine characterization of new antibodies against rhamnogalacturonan I. WallTraC Symposiu

Biology-driven, microarray-assisted selection of cell wall directed antibodies

Biology-driven, microarray-assisted selection of cell wall directed antibodies. Gordon Research Conference on Plant Cell Wall

Development of novel monoclonal antibodies against starch and ulvan - implications for antibody production against polysaccharides with limited immunogenicity

Monoclonal antibodies (mAbs) are widely used and powerful research tools, but the generation of mAbs against glycan epitopes is generally more problematic than against proteins. This is especially significant for research on polysaccharide-rich land plants and algae (Viridiplantae). Most antibody production is based on using single antigens, however, there are significant gaps in the current repertoire of mAbs against some glycan targets with low immunogenicity. We approached mAb production in a different way and immunised with a complex mixture of polysaccharides. The multiplexed screening capability of carbohydrate microarrays was then exploited to deconvolute the specificities of individual mAbs. Using this strategy, we generated a set of novel mAbs, including one against starch (INCh1) and one against ulvan (INCh2). These polysaccharides are important storage and structural polymers respectively, but both are generally considered as having limited immunogenicity. INCh1 and INCh2 therefore represent important new molecular probes for Viridiplantae research. Moreover, since the alpha-(1-4)-glucan epitope recognised by INCh1 is also a component of glycogen, this mAb can also be used in mammalian systems. We describe the detailed characterisation of INCh1 and INCh2, and discuss the potential of a non-directed mass-screening approach for mAb production against some glycan targets

Complexity of the Ruminococcus flavefaciens cellulosome reflects an expansion in glycan recognition

International audienceno abstrac