Extraction, texture analysis and polysaccharide epitope mapping data of sequential extracts of strawberry, apple, tomato and aubergine fruit parenchyma

The data included in this article are related to the research article entitled “Disentangling pectic homogalacturonan and rhamnogalacturonan-I polysaccharides: evidence for sub-populations in fruit parenchyma systems” (Cornuault et al., 2018) [1]. Cell wall properties are an important contributor to fruit texture. These datasets compile textural and immunochemical analysis of polysaccharides of four economically important fruit crops: tomato, strawberry, aubergine and apple with contrasting textures and related taxonomical origins. Cell wall components and their extractability were assessed using characterized monoclonal antibodies. In addition, textural data obtained for the four parenchyma systems show variations in the mechanical properties. The two datasets are a basis to relate cell wall composition and organization to the mechanical properties of the fruit parenchyma tissues

Branched Pectic Galactan in Phloem-Sieve-Element Cell Walls: Implications for Cell Mechanics

A major question in plant biology concerns the specification and functional differentiation of cell types. This is in the context of constraints imposed by networks of cell walls that both adhere cells and contribute to the form and function of developing organs. Here, we report the identification of a glycan epitope that is specific to phloem sieve element cell walls in several systems. A monoclonal antibody, designated LM26, binds to the cell wall of phloem sieve elements in stems of Arabidopsis (Arabidopsis thaliana), Miscanthus x giganteus, and notably sugar beet (Beta vulgaris) roots where phloem identification is an important factor for the study of phloem unloading of Suc. Using microarrays of synthetic oligosaccharides, the LM26 epitope has been identified as a β-1,6-galactosyl substitution of β-1,4-galactan requiring more than three backbone residues for optimized recognition. This branched galactan structure has previously been identified in garlic (Allium sativum) bulbs in which the LM26 epitope is widespread throughout most cell walls including those of phloem cells. Garlic bulb cell wall material has been used to confirm the association of the LM26 epitope with cell wall pectic rhamnogalacturonan-I polysaccharides. In the phloem tissues of grass stems, the LM26 epitope has a complementary pattern to that of the LM5 linear β-1,4-galactan epitope, which is detected only in companion cell walls. Mechanical probing of transverse sections of M. x giganteus stems and leaves by atomic force microscopy indicates that phloem sieve element cell walls have a lower indentation modulus (indicative of higher elasticity) than companion cell walls

Monoclonal antibodies directed to fucoidan preparations from brown algae

Cell walls of the brown algae contain a diverse range of polysaccharides with useful bioactivities. The precise structures of the sulfated fucan/fucoidan group of polysaccharides and their roles in generating cell wall architectures and cell properties are not known in detail. Four rat monoclonal antibodies, BAM1 to BAM4, directed to sulfated fucan preparations, have been generated and used to dissect the heterogeneity of brown algal cell wall polysaccharides. BAM1 and BAM4, respectively, bind to a non-sulfated epitope and a sulfated epitope present in the sulfated fucan preparations. BAM2 and BAM3 identified additional distinct epitopes present in the fucoidan preparations. All four epitopes, not yet fully characterised, occur widely within the major brown algal taxonomic groups and show divergent distribution patterns in tissues. The analysis of cell wall extractions and fluorescence imaging reveal differences in the occurrence of the BAM1 to BAM4 epitopes in various tissues of Fucus vesiculosus. In Ectocarpus subulatus, a species closely related to the brown algal model Ectocarpus siliculosus, the BAM4 sulfated epitope was modulated in relation to salinity levels. This new set of monoclonal antibodies will be useful for the dissection of the highly complex and yet poorly resolved sulfated polysaccharides in the brown algae in relation to their ecological and economic significance

Molecular and biochemical tool for the analysis of glycans in plant cell walls

Molecular and biochemical tool for the analysis of glycans in plant cell walls. WallTraC Symposiu

Production and fine characterisation of new monoclonal antibodies against rhamnogalacturonan I

National audienceThe functional role of the complex pectic rhamnogalacturonane I (RGI) in the context of cell biology is still unclear. RGI is mainly composed of a repeating disaccharide unit [,2)-a-L-rhamnosep-(1,4)-a-D-galacturonic acidp-(1,]n (RU)n decorated primarily with arabinan and (arabino)-galactan side-chains. Monoclonal antibodies (mAbs) are useful to probe pectin structural domains in situ. Several mAbs to the RGI backbone and to arabinan and (arabino)-galactan side-chains have been developed but up to now, there is no mAb against rhamnogalacturonan stretches bearing arabinose or galactose units. Here, we report on the production and fine characterisation of new mAbs against RGI side chains connected to the rhamnogalacturonan backbone. The study first focused on the generation of adequate antigens. A pool of low-branched RU oligosaccharides was prepared and purified from potato pulp. mAbs were raised against low-branched RU oligosaccharides-ovalbumin conjugates. Promising clones producing mAbs recognizing low-branched RU oligosaccharides but recognizing neither unbranched RU oligosaccharides nor galactan oligosaccharides were recovered. The combination of glycan microarray and sub-fractionation of the low-branched RU oligosaccharides pool by anion exchange chromatography prior to competitive ELISA studies allowed fine mAbs characterisation. The use of these new mAbs in immunocytochemistry is expected to give a better understanding of RGI role in planta

Production and fine characterization of new antibodies against rhamnogalacturonan I

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

The deconstruction of pectic rhamnogalacturonan I unmasks the occurrence of a novel arabinogalactan oligosaccharide epitope

Rhamnogalacturonan I (RGI) is a pectic polysaccharide composed of a backbone of alternating rhamnose and galacturonic acid residues with side chains containing galactose and/or arabinose residues. The structure of these side chains and the degree of substitution of rhamnose residues are extremely variable and depend on species, organs, cell types and developmental stages. Deciphering RGI function requires extending the current set of monoclonal antibodies (mAbs) directed to this polymer. Here, we describe the generation of a new mAb that recognizes a heterogeneous subdomain of RGI. The mAb, INRA-AGI-1, was produced by immunization of mice with RGI oligosaccharides isolated from potato tubers. These oligomers consisted of highly branched RGI backbones substituted with short side chains. INRA-AGI-1 bound specifically to RGI isolated from galactan-rich cell walls and displayed no binding to other pectic domains. In order to identify its RGI-related epitope, potato RGI oligosaccharides were fractionated by anion-exchange chromatography. Antibody recognition was assessed for each chromatographic fraction. INRA-AGI-1 recognizes a linear chain of (1→4)-linked galactose and (1→5)-linked arabinose residues. By combining the use of INRA-AGI-1 with LM5, LM6 and INRA-RU1 mAbs and enzymatic pre-treatments, evidence is presented of spatial differences in RGI motif distribution within individual cell walls of potato tubers and carrot roots. These observations raise questions about the biosynthesis and assembly of pectin structural domains and their integration and remodeling in cell walls.fals