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Localization of xyloglucan epitopes in tobacco stems. Immuno- fluorescence imaging of transverse sections of tobacco stem pith parenchyma cell walls with anti-xyloglucan mAb after a pectate lyase pretreatment is shown. A , Calcofluor fluorescence showing all cell walls. B , the same section as A with immunofluorescence labeling with mAb LM15 showing abundant binding to cell walls at the corners of intercellular spaces (*). C , equivalent section showing immunofluorescence labeling with mAb LM24 indicating most abundant labeling in regions of adhered cell walls between intercellular spaces. D , equivalent section showing immunofluorescence labeling with mAb LM25, which binds abundantly to cell walls lining intercellular spaces. Scale bars ϭ 10 m.
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Microarrays are powerful tools for high throughput analysis, and hundreds or thousands of molecular interactions can be assessed
simultaneously using very small amounts of analytes. Nucleotide microarrays are well established in plant research, but carbohydrate
microarrays are much less established, and one reason for this is a lack of suitable gly...
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Context 1
... oligomers (and in the case of LM25 weak binding to unsubstituted -glucan) than the previously characterized mAb LM15 (38) (Fig. 3J). The differences in the anti-xyloglucan mAb array binding profiles were reflected in differing binding profiles when applied to pectate lyase-treated transverse sections of tobacco stem pith parenchyma as shown in Fig. 4. Previous work had demonstrated that after pectic homogalacturonan removal, the LM15 xyloglucan epitope is revealed abundantly at the corners of intercellular spaces (Marcus et al. (38)) (Fig. 4B). In equivalent material, the LM24 epitope was most abundant in adhered cell walls between inter- cellular spaces (Fig. 4C), and the LM25 ...
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... were reflected in differing binding profiles when applied to pectate lyase-treated transverse sections of tobacco stem pith parenchyma as shown in Fig. 4. Previous work had demonstrated that after pectic homogalacturonan removal, the LM15 xyloglucan epitope is revealed abundantly at the corners of intercellular spaces (Marcus et al. (38)) (Fig. 4B). In equivalent material, the LM24 epitope was most abundant in adhered cell walls between inter- cellular spaces (Fig. 4C), and the LM25 epitope was localized in cell walls lining intercellular spaces (Fig. ...
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... pith parenchyma as shown in Fig. 4. Previous work had demonstrated that after pectic homogalacturonan removal, the LM15 xyloglucan epitope is revealed abundantly at the corners of intercellular spaces (Marcus et al. (38)) (Fig. 4B). In equivalent material, the LM24 epitope was most abundant in adhered cell walls between inter- cellular spaces (Fig. 4C), and the LM25 epitope was localized in cell walls lining intercellular spaces (Fig. ...
Context 4
... homogalacturonan removal, the LM15 xyloglucan epitope is revealed abundantly at the corners of intercellular spaces (Marcus et al. (38)) (Fig. 4B). In equivalent material, the LM24 epitope was most abundant in adhered cell walls between inter- cellular spaces (Fig. 4C), and the LM25 epitope was localized in cell walls lining intercellular spaces (Fig. ...
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In nature, carbohydrates form an important family of biomolecules, as simple or complex carbohydrates, either alone or covalently linked to proteins or lipids. Most of the earlier studies on carbohydrates focused on plant polysaccharides, such as cellulose, starch, pectins, and so on, largely because of their wide range of applications. More recent...
Rhamnogalacturonans I represent a group of plant cell wall polysaccharides having the most complex organization and variable structure. These polymers, combined in one group due to the presence of a backbone composed of alternating [→4)-α-d-GalpA-(1→2)-α-l-Rhap(1→] dimers, can occur in the cell wall both as parts of a pectin complex and by themselv...
Citations
... SM, soluble mucilage; AM, adherent mucilage; DS, demucilaged seed; Fuc, fucose; Rha, rhamnose; Ara, Arabinose; Gal, galactose; Glc, glucose; Man, mannose; Xyl, xylose; GalA, galacturonic acid; GlcA, glucuronic acid. (Voiniciuc et al., 2015a) and we observed a considerable amount of Xyl and Glc in our monosaccharide analysis, we employed LM25 (XXXG, XXLG, XLLG) to detect the presence of different HC epitopes (Pedersen et al., 2012). An intense label was observed across all tested dilutions, strongly indicating the presence of xyloglucan. ...
Chilean papaya, also known as mountain papaya (Vasconcellea pubescens), is a fruit valued for its nutritional value and pleasant fragrance. The oblong fruit, featuring five ridges and a seed-filled mucilage cavity, is typically consumed cooked due to its high protease content. The mucilage and the seeds are usually discarded as byproducts. This study analyzed the biochemical composition of mountain papaya seed mucilage using methods such as HPAEC and immunolabeling. Results revealed that papaya seeds yield nearly 20% of their weight in mucilage polysaccharides, which can be separated into soluble and adherent layers. The mucilage exhibited a high proportion of acidic sugars, indicating that homogalacturonan (HG) is the predominant domain. It also contained other domains like rhamnogalacturonan-I (RG-I) and hemicelluloses, predominantly xyloglucan. The HG-rich mucilage, currently considered waste, emerges as a promising source of polysaccharides, indicating its multifaceted utility in various industrial applications.
... These antibodies identified epitopes associated with β-1,6galactosyl substitution on β-1,4-galactan and galactosylated xyloglucan. This recognition suggests that these polysaccharides contain β-glucan epitopes [29][30][31]. NaOH and cellulase treatments predominantly extracted polysaccharides likely recognised by LM6 and LM2 antibodies. This suggests the presence of (1→5)-α-L-arabinan and Arabinogalactan protein (AGP) structures featuring β-linked glycosylglucopyranuronic acid (GlcA) [32][33][34]. ...
Banana peel (BP) is the primary by-product generated during banana processing which causes numerous environmental issues. This study examines the physical attributes, proximate analysis, glycoarray profiling, antioxidant abilities, and prebiotic activity of BP. The analysis demonstrated that carbohydrates constituted the primary components of BP and the glycoarray profiling indicated that BP contains multiple pectin and hemicellulose structures. BP also contained phenolic compounds, including (+)-catechin and gallic acid, flavonoid compounds, and antioxidant activities. BP demonstrated prebiotic effects by promoting the proliferation of advantageous gut bacteria while inhibiting the growth of harmful bacteria. The prebiotic index scores demonstrated that BP exhibited a greater capacity to promote the growth of beneficial bacteria in comparison to regular sugar. The study demonstrated the potential of the BP as a valuable source of dietary fibre, bioactive compounds, and prebiotics. These components have beneficial characteristics and can be utilised in the production of food, feed additives, and functional food.
... Each sample was spotted with four fivefold dilutions and two technical replicates. The arrays were probed with antibodies listed in Table 1 and detected using anti-rat or anti-mouse (BioSupplies) secondary antibodies conjugated to alkaline phosphatase (Sigma-Aldrich) as described in Pedersen et al. (2012). The microarrays were developed using an NBT/BCIP substrate containing nitroblue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolylphosphate p-toluidine salt (BCIP) in alkaline phosphatase buffer (100 mM NaCl, 5 mM MgCl 2 , 100 mM diethanol-amine, pH 9.5). ...
The evolution of land flora was an epochal event in the history of planet Earth. The
success of plants, and especially flowering plants, in colonizing all but the most
hostile environments required multiple mechanisms of adaptation. The mainly
polysaccharide‐based cell walls of flowering plants, which are indispensable for
water transport and structural support, are one of the most important adaptations to
life on land. Thus, development of vasculature is regarded as a seminal event in cell
wall evolution, but the impact of further refinements and diversification of cell wall
compositions and architectures on radiation of flowering plant families is less well
understood. We approached this from a glyco‐profiling perspective and, using
carbohydrate microarrays and monoclonal antibodies, studied the cell walls of
287 plant species selected to represent important evolutionary dichotomies and
adaptation to a variety of habitats. The results support the conclusion that radiation
of flowering plant families was indeed accompanied by changes in cell wall fine
structure and that these changes can obscure earlier evolutionary events.
Convergent cell wall adaptations identified by our analyses do not appear to be
associated with plants with similar lifestyles but that are taxonomically distantly
related. We conclude that cell wall structure is linked to phylogeny more strongly
than to habitat or lifestyle and propose that there are many approaches of
adaptation to any given ecological niche.
... In this study, the following rat monoclonal antibodies (MAbs) were used: LM25, which binds to xyloglucan [19], LM5, which binds to pectic galactan [20], LM6, which binds to pectic arabinan [21], LM19, which binds to unesterified pectic HG [22], JIM7, which binds to partially methyl-esterified pectic HG [22], and LM20, which binds to highly methyl-esterified pectic HG [22]. ...
The blackening of cut carrots causes substantial economic losses to the food industry. Blackening was not observed in carrots that had been stored underground for less than a year, but the susceptibility to blackening increased with the age of the carrots that were stored underground for longer periods. Samples of black, border, and orange tissues from processed carrot batons and slices, prepared under industry standard conditions, were analyzed to identify the molecular and metabolic mechanisms underpinning processing-induced blackening. The black tissues showed substantial molecular and metabolic rewiring and large changes in the cell wall structure, with a decreased abundance of xyloglucan, pectins (homogalacturonan, rhamnogalacturonan-I, galactan and arabinan), and higher levels of lignin and other phenolic compounds when compared to orange tissues. Metabolite profiling analysis showed that there was a major shift from primary to secondary metabolism in the black tissues, which were depleted in sugars, amino acids, and tricarboxylic acid (TCA) cycle intermediates but were rich in phenolic compounds. These findings suggest that processing triggers a release from quiescence. Transcripts encoding proteins associated with secondary metabolism were less abundant in the black tissues, but there were no increases in transcripts associated with oxidative stress responses, programmed cell death, or senescence. We conclude that restraining quiescence release alters cell wall metabolism and composition, particularly regarding pectin composition, in a manner that increases susceptibility to blackening upon processing.
... ND: not detected. References [70][71][72][73][74][75][76][77][78][79][80] are cited in the supplementary materials. Data Availability Statement: NMR data and metadata have been deposited in the recherche.data.gouv.fr ...
There is a growing demand for molecules of natural origin for biocontrol and biostimulation, given the current trend away from synthetic chemical products. Leachates extracted from plantain stems were obtained after biodegradation of the plant material. To characterize the leachate, quantitative determinations of nitrogen, carbon, phosphorus, and cations (K+, Ca2+, Mg2+, Na+), Q2/4, Q2/6, and Q4/6 absorbance ratios, and metabolomic analysis were carried out. The potential role of plantain leachates as fungicide, elicitor of plant defense, and/or plant biostimulant was evaluated by agar well diffusion method, phenotypic, molecular, and imaging approaches. The plant extracts induced a slight inhibition of fungal growth of an aggressive strain of Colletotrichum gloeosporioides, which causes anthracnose. Organic compounds such as cinnamic, ellagic, quinic, and fulvic acids and indole alkaloid such as ellipticine, along with some minerals such as potassium, calcium, and phosphorus, may be responsible for the inhibition of fungal growth. In addition, jasmonic, benzoic, and salicylic acids, which are known to play a role in plant defense and as biostimulants in tomato, were detected in leachate extract. Indeed, foliar application of banana leachate induced overexpression of LOXD, PPOD, and Worky70-80 genes, which are involved in phenylpropanoid metabolism, jasmonic acid biosynthesis, and salicylic acid metabolism, respectively. Leachate also activated root growth in tomato seedlings. However, the main impact of the leachate was observed on mature plants, where it caused a reduction in leaf area and fresh weight, the remodeling of stem cell wall glycopolymers, and an increase in the expression of proline dehydrogenase.
... Xyloglucan, which together with cellulose is thought to be involved in the formation of 'biomechanical hotspots' and the realization of expansin action (Park & Cosgrove, 2012), was detected in the early elongation zone of maize, rye and soybean roots using two antibodies-LM25 for xyloglucan (Pedersen et al., 2012) and CCRC-M1 for fucosylated xyloglucan (Puhlmann et al., 1994) (Figure 4). ...
Plant growth and morphogenesis are determined by the mechanical properties of its cell walls. Using atomic force microscopy, we have characterized the dynamics of cell wall elasticity in different tissues in developing roots of several plant species. The elongation growth zone of roots of all species studied was distinguished by a reduced modulus of elasticity of most cell walls compared to the meristem or late elongation zone. Within the individual developmental zones of roots, there were also significant differences in the elasticity of the cell walls of the different tissues, thus identifying the tissues that limit root growth in the different species. In cereals, this is mainly the inner cortex, whereas in dicotyledons this function is performed by the outer tissues-rhizodermis and cortex. These differences result in a different behaviour of the roots of these species during longitudinal dissection. Modelling of longitudinal root dissection using measured properties confirmed the difference shown. Thus, the morphogenesis of monocotyledonous and dicotyledonous roots relies on different tissues as growth limiting, which should be taken into account when analyzing the localization of associated molecular events. At the same time, no matrix polysaccharide was found whose immunolabelling in type I or type II cell walls would predict their mechanical properties. However, assessment of the degree of anisotropy of cortical microtubules showed a striking correlation with the elasticity of the corresponding cell walls in all species studied.
... These antibodies detect the epitopes of the non-reducing end of (1 ! 4)-β-D-xylan (LM10; McCartney et al., 2005; Ruprecht et al., 2017), as well as the XXXG-motif of xyloglucan (LM15;Marcus et al., 2008;Pedersen et al., 2012). The highest absorption with both antibodies was detectable in C. globularis, followed by C. tomentosa and C. subspinosa. ...
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.
... The primary monoclonal antibodies (PlantProbes, Leeds, UK) were used at a dilution of 1:10 in 10 mM PBS: LM7 (anti-homogalacturonan) [13], LM8 (anti-xylogalacturonan) [14], LM19 (anti-homogalacturonan) [15], LM20 (anti-homogalacturonan) [15] and 2F4 for dimeric associations of Ca with homogalacturonan [16]. Those reacting with hemicelluloses were: LM11 (anti-xylan/arabinoxylan) [17], LM21 (anti-mannan) [18] and LM25 (antixyloglucan) [19]. Only the mAB 2F4 was buffered in 20 mM Tris(hydroxymethyl)methylamin buffer (TRIS-HCl) adjusted to pH 8.2. ...
... Vascular browning and cavities in´Nicoter´apples [19] (K). Cavities (ca) indicated by white arrows; pi, pith; rp, ray parenchyma; vb, vascular bundle. ...
... The antibodies against pectins were: LM7 (anti-homogalacturonan) [13], LM8 (anti-xylogalacturonan) [14], LM19 (anti-homogalacturonan) [15], LM20 (anti-homogalacturonan) [15] and 2F4 for dimeric associations of Ca with homogalacturonan [16]. Those reacting with hemicelluloses were: LM11 (anti-xylan/arabinoxylan) [17], LM21 (anti-mannan) [18] and LM25 (anti-xyloglucan) [19]. The GFP fluorescence was rated on a three-point scale: 0 no fluorescence, + some cell walls fluorescing, ++ all cell walls fluorescing; n = 3. ...
'Nicoter' apples (Malus × domestica Borkh.) occasionally develop a disorder referred to as vascular browning. Symptomatic fruit are perceived as being of low quality. The objective was to identify the mechanistic basis of this disorder. The frequency of symptomatic 'Nicoter' apples differed between growing sites and increased with delayed harvest. Typical symptoms are tissue browning and cavities in the ray parenchyma of the calyx region, and occasionally also of the stem end. Cavity size is positively correlated with the extent of tissue browning. Cavities were oriented radially in the direction of the bisecting line between the radii connecting the calyx/pedicel axis to the sepal and petal bundles. Microscopy revealed cell wall fragments in the cavities indicating physical rupture of cell walls. Immunolabelling of cell wall epitopes offered no evidence for separation of cells along cell walls. The growth pattern in 'Nicoter' is similar to that in its parents 'Gala' and 'Braeburn'. Allometric analyses revealed no differences in growth in fruit length among the three cultivars. However, the allometric analyses of growth in diameter revealed a marked increase in the distance between the surface of the calyx cavity and the vascular bundle in 'Nicoter', that was absent in 'Braeburn' and 'Gala'. This increase displaced the petal bundles in the ray parenchyma outwards and subjected the tissue between the petal and sepal bundles to tangential strain. Rupture of cells results in tissue browning and cavity formation. A timely harvest is a practicable countermeasure for decreasing the incidence of vascular browning.
... In this study, the following rat monoclonal antibodies (MAbs) were used: LM25 which binds to xyloglucan [18], LM5 which binds to pectic galactan [19], LM6 to pectic arabinan [20], LM19 to unesterified pectic HG [21], JIM7 to partially methyl-esterified pectic HG [21] and LM20 to highly methyl-esterified pectic HG [21]. ...
The blackening of cut carrots decreases their shelf life and causes severe economic losses but the molecular and metabolic mechanisms that underpin this phenomenon remain poorly characterized. Studies were therefore undertaken to determine the molecular and metabolic causes of the blackening. The susceptibility of blackening was dependent on the period of time that the crop was stored underground prior to harvest. The structure of the cell walls in the black regions was substantially changed compared to the orange regions. The black regions of carrot batons had decreased immunodetection of xyloglucan, HG-pectin, RG-I pectin, galactan and arabinan but had higher levels of lignin and phenolic compounds compared to the orange regions. Transcript profiling analysis revealed that phytohormone signalling processes were activated in the black regions. Transcripts associated with auxin signalling and ethylene-responsive transcription factors were increased in the black regions. In contrast, the levels of transcripts encoding proteins associated with secondary metabolism were decreased in the black regions. These findings implicate ethylene and auxin-related processes in the control of the primary to secondary metabolism shift that results in lignification and cell wall disruption that underpin the blackening process.
... In this study, the following rat monoclonal antibodies (MAbs) were used: LM25 which binds to xyloglucan [18], LM5 which binds to pectic galactan [19], LM6 to pectic arabinan [20], LM19 to unesterified pectic HG [21], JIM7 to partially methyl-esterified pectic HG [21] and LM20 to highly methyl-esterified pectic HG [21]. ...
The blackening of cut carrots decreases their shelf life and causes severe economic losses but the molecular and metabolic mechanisms that underpin this phenomenon remain poorly characterized. Studies were therefore undertaken to determine the molecular and metabolic causes of the blackening. The susceptibility of blackening was dependent on the period of time that the crop was stored underground prior to harvest. The structure of the cell walls in the black regions was substantially changed compared to the orange regions. The black regions of carrot batons had decreased immunodetection of xyloglucan, HG-pectin, RG-I pectin, galactan and arabinan but had higher levels of lignin and phenolic compounds compared to the orange regions. Transcript profiling analysis revealed that phytohormone signalling processes were activated in the black regions. Transcripts associated with auxin signalling and ethylene-responsive transcription factors were increased in the black regions. In contrast, the levels of transcripts encoding proteins associated with secondary metabolism were decreased in the black regions. These findings implicate ethylene and auxin-related processes in the control of the primary to secondary metabolism shift that results in lignification and cell wall disruption that underpin the blackening process.
