Contributions to Arabinogalactan Protein Analysis

Castilleux, Romain; Ropitaux, Marc; Manasfi, Youssef; Bernard, Sophie; Vicré-Gibouin, Maïté; Driouich, Azeddine

doi:10.1007/978-1-0716-0621-6_22

Cited by 6 publications

(4 citation statements)

References 33 publications

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“…Detection of AGPs on agarose gel was performed according to Castilleux et al (2020) . Water-extracted AGPs from 15 mg pooled samples were loaded onto a 1% (w/v) agarose gel containing 90 mM Tris base pH 8.3 with HCl, 90 mM boric acid, and 2 mM Na 2 EDTA (H 2 O) 2 , and run at 100 V for 1 h. β-D-glucosyl Yariv was synthesized in house according to the protocol by Yariv et al (1962) .…”

Section: Methodsmentioning

confidence: 99%

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

et al. 2020

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Plant cell wall associated hydroxyproline-rich glycoproteins (HRGPs) are involved in several aspects of plant growth and development, including wood formation in trees. HRGPs such as arabinogalactan-proteins (AGPs), extensins (EXTs), and proline rich proteins (PRPs) are important for the development and architecture of plant cell walls. Analysis of publicly available gene expression data revealed that many HRGP encoding genes show tight spatio-temporal expression patterns in the developing wood of Populus that are indicative of specific functions during wood formation. Similar results were obtained for the expression of glycosyl transferases putatively involved in HRGP glycosylation. In situ immunolabelling of transverse wood sections using AGP and EXT antibodies revealed the cell type specificity of different epitopes. In mature wood AGP epitopes were located in xylem ray cell walls, whereas EXT epitopes were specifically observed between neighboring xylem vessels, and on the ray cell side of the vessel walls, likely in association with pits. Molecular mass and glycan analysis of AGPs and EXTs in phloem/cambium, developing xylem, and mature xylem revealed clear differences in glycan structures and size between the tissues. Separation of AGPs by agarose gel electrophoresis and staining with β-D-glucosyl Yariv confirmed the presence of different AGP populations in phloem/cambium and xylem. These results reveal the diverse changes in HRGP-related processes that occur during wood formation at the gene expression and HRGP glycan biosynthesis levels, and relate HRGPs and glycosylation processes to the developmental processes of wood formation.

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Section: Methodsmentioning

confidence: 99%

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

et al. 2020

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“…For rapid detection and semi-quantification of AGPs, the radial gel diffusion assay using Yariv specific reactivity was carried out as in Holst and Clarke [27] and Castilleux [28]. The samples, i.e.…”

Section: Yariv Reagent Reactivity -Radial Gel Diffusion Assaymentioning

confidence: 99%

Working towards arabinogalactan proteins (AGPs) from fruit: carbohydrate composition and impact on fungal growth

et al. 2022

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Background Arabinogalactan proteins (AGPs) are extracellular matrix constituents involved in plant response to fungal infection. The aim of the current study was to investigate the antifungal effect of AGPs ex situ and to determine the structural features of AGPs that may have an influence on this activity. The features of AGPs isolated from fruit were investigated with molecular tools based on specific monoclonal antibodies recognizing carbohydrate AGP epitopes. The Antifungal (well-diffusion) Susceptibility Test and the Agar Invasion Test were used to assess the impact of AGPs on Penicillium notatum culture. Results The results definitely ruled out the influence of AGPs on fungal growth. The immunochemical analyses revealed that AGPs consist mainly of carbohydrate chains composed of β-linked glucuronosyl residues recognized by LM2 and GlcA-β(1 → 3)-GalA-α(1 → 2) Rha recognized by JIM13, which do not have the same functional properties outside the plant cell in in vitro experimental conditions. Conclusions The action of a single cell wall component does not elicit any influence ex situ. The extensive accumulation of glycan chains of AGPs in infected tissue as a result of a complex mechanism occurring in the cell wall emphasizes the importance of dependencies between particular components of the extracellular matrix in response to fungal attack.

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“…The root cap exhibits high secretory activity, as evidenced by the presence of hypertrophied Golgi stacks (Whaley et al, 1959;Spink and Wilson, 1968;Staehelin et al, 1990). The mucilage of the root cap primarily comprises pectin polysaccharides and proteoglycans (Read et al, 1999;Maeda et al, 2019;Castilleux et al, 2020;Ropitaux et al, 2020). Golgi stacks in border cells/BLCs and peripheral cells, precursors of border cells/BLCs have swollen cisternae where synthesis of pectin and addition of polysaccharide moieties of proteoglycans occur (Wang et al, 2017;Wang and Kang, 2018).…”

Section: Introductionmentioning

confidence: 99%

Bearskin2 mediates the coordinated secretion of xylogalacturonan and root cap polygalacturonase in Arabidopsis border-like cells

Liu

Wang

Goh

et al. 2023

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Border-like cells (BLCs) are sheets of cells that are continuously sloughed off and replenished at the Arabidopsis root cap surface. ROOT CAP POLYGALACTURONASE (RCPG) encodes a putative pectinase involved in BLC shedding. Xylogalacturonan (XGA) is a pectic polysaccharide whose synthesis is associated with cell detachment and secreted separately from other cell wall polysaccharides. BEARSKIN1 (BRN1) and BRN2 are Arabidopsis NAC family transcription factors, and RCPG expression is inhibited in brn1/2. To explore the link between XGA and RCPG, we examined XGA synthesis in Arabidopsis lines with altered RCPG levels. We found that RCPG was contained in XGA-carrying vesicles budding from the trans-Golgi, but XGA synthesis was not affected in the rcpg mutant. XGA was absent in BLCs of brn2, but not of brn1, indicating that BRN2 is necessary for XGA synthesis. Overexpression of functional RCPG-GFP (oeRCPG-GFP) caused upregulation of BRN2, ectopic XGA synthesis, overaccumulation of endogenous RCPG, and accelerated BLC turnover, suggesting a positive regulatory loop between RCPG and BRN2. Inactivation of BRN2 in oeRCPG-GFP suppressed RCPG-GFP expression, excess RCPG, and XGA synthesis . Our data provide evidence that XGA and RCPG are secreted together and that BRN2 controls XGA synthesis, which facilitates RCPG export and BLC separation.

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Contributions to Arabinogalactan Protein Analysis

Cited by 6 publications

References 33 publications

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

The Spatio-Temporal Distribution of Cell Wall-Associated Glycoproteins During Wood Formation in Populus

Working towards arabinogalactan proteins (AGPs) from fruit: carbohydrate composition and impact on fungal growth

Bearskin2 mediates the coordinated secretion of xylogalacturonan and root cap polygalacturonase in Arabidopsis border-like cells

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