Isolation and characterization of cell walls from the mesocarp of mature grape berries (Vitis vinifera)

Nunan, Kylie J.; Sims, Ian M.; Bacic, Antony; Robinson, Simon P.

doi:10.1007/s00050169

Cited by 23 publications

(35 citation statements)

References 38 publications

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“…Methylation and reduction of the radiolabeled oligosaccharides were performed as described previously (Nunan et al, 1997). Briefly, collected radiolabeled peaks from four reactions were methylated using NaOH/CH 3 I and were hydrolyzed using 2.5 m TFA for 4 h at 100°C.…”

Section: Analysis Of Radiolabeled Productsmentioning

confidence: 99%

Solubilization of an Arabinan Arabinosyltransferase Activity from Mung Bean Hypocotyls

Nunan¹,

Scheller²

2003

Plant Physiology

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The biosynthesis of polysaccharides destined for the plant cell wall and the subsequent assembly of the cell wall are poorly understood processes that are currently the focus of much research. Arabinan, a component of the pectic polysaccharide rhamnogalacturonan I, is composed of arabinosyl residues connected via various glycosidic linkages, and therefore, the biosynthesis of arabinan is likely to involve more than one arabinosyltransferase. We have studied the transfer of [ 14 C]arabinose (Ara) from UDP-l-arabinopyranose onto polysaccharides using microsomal membranes isolated from mung bean (Vigna radiata) hypocotyls. [ 14 C]arabinosyl and [ 14 C]xylosyl residues were incorporated into endogenous products due to the presence of UDP-Xyl-4-epimerase activity. Enzymatic digestion of endogenous products with endo-arabinanase released very little radiolabeled sugars, whereas digestion with arabinofuranosidase released some [ 14 C]Ara. Microsomal membranes solubilized with the detergent octyl glucoside were able to add a single [ 14 C]Ara residue onto (135)-linked ␣-l-arabino-oligosaccharide acceptors. The reaction had a pH optimum of 6.5 and a requirement for manganese ions. However, enzymatic digestion of the radiolabeled oligosaccharides with endo-arabinanase and arabinofuranosidases could not fully release the radiolabeled Ara residue, indicating that the [ 14 C]Ara residue was not a (132)-, (133)-, or (135)-linked ␣-l-arabinofuranosyl residue. Rather, mild acid treatment of the product suggested that the radiolabeled Ara residue was in a pyranose conformation, and this result was confirmed by thin-layer chromatography of radiolabeled partially methylated sugars. Using microsomal membranes separated on a discontinuous sucrose gradient, the arabinosyltransferase activity appears to be mainly localized to Golgi membranes.Pectic polysaccharides are major components of primary plant cell walls and middle lamella. Three main types of pectic polysaccharides have been identified, homogalacturonan, rhamnogalacturonan I (RG I), and rhamnogalacturonan II (RG II). RG I is composed of a backbone of alternating [34-␣-d-GalA-(132)-␣-l-Rha- (13] in which some of the rhamnose residues are substituted with complex side chains containing Ara and Gal residues. The composition and size of these side chains varies depending on the plant species, tissue type, and stage of development (O'Neill et al., 1990;Carpita and Gibeaut, 1993). In general, the arabinan side chains consist of a linear chain of (135)-linked ␣-l-arabinofuranosyl residues that may be substituted at the O-3 and/or O-2 with additional arabinofuranosyl residues (Carpita and Gibeaut, 1993;Schols and Voragen, 2002).The study of the biosynthesis of plant cell wall polysaccharides in plants is a growing field, but to date, very few genes encoding glycosyltransferases have been characterized (Edwards et al., 1999;Perrin et al., 1999;Faik et al., 2002). Notably, many different transferases must be involved in pectin biosynthesis, but until very recently, none had been isola...

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Section: Analysis Of Radiolabeled Productsmentioning

confidence: 99%

Solubilization of an Arabinan Arabinosyltransferase Activity from Mung Bean Hypocotyls

Nunan¹,

Scheller²

2003

Plant Physiology

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“…During thawing of frozen berries, skin and seeds were removed and mesocarp cell walls were isolated as described by Nunan et al (1997). Skin and seeds were removed manually and the remaining mesocarp tissue was homogenized in 4 volumes of absolute ethanol using a household blender.…”

Section: Preparation Of Cell Wallsmentioning

confidence: 99%

“…Protein in the wall preparations and amino acid compositions were determined as described by Nunan et al (1997). Hyp in hydrolysates was measured colorimetrically by the procedure of Kivirikko and Liesmaa (1959).…”

Section: Protein and Amino Acid Estimationsmentioning

confidence: 99%

“…Linkage analyses of cell wall preparations and extracted wall fractions were performed as described in Nunan et al (1997). All methylation analyses were performed in duplicate, and where significant variation was observed, the analyses were repeated again in duplicate.…”

Section: Polysaccharide Linkage Analysismentioning

confidence: 99%

“…In the present study a recently developed procedure for the preparation of cell walls from the mesocarp of grape berries (Nunan et al, 1997) has been used to isolate walls at various stages during berry development and ripening. The appearance of the walls during ripening has been examined by light and fluorescence microscopy.…”

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Changes in Cell Wall Composition during Ripening of Grape Berries

et al. 1998

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Cell walls were isolated from the mesocarp of grape (Vitis vinifera L.) berries at developmental stages from before veraison through to the final ripe berry. Fluorescence and light microscopy of intact berries revealed no measurable change in cell wall thickness as the mesocarp cells expanded in the ripening fruit. Isolated walls were analyzed for their protein contents and amino acid compositions, and for changes in the composition and solubility of constituent polysaccharides during development. Increases in protein content after veraison were accompanied by an approximate 3-fold increase in hydroxyproline content. The type I arabinogalactan content of the pectic polysaccharides decreased from approximately 20 mol % of total wall polysaccharides to about 4 mol % of wall polysaccharides during berry development. Galacturonan content increased from 26 to 41 mol % of wall polysaccharides, and the galacturonan appeared to become more soluble as ripening progressed. After an initial decrease in the degree of esterification of pectic polysaccharides, no further changes were observed nor were there large variations in cellulose (30-35 mol % of wall polysaccharides) or xyloglucan (approximately 10 mol % of wall polysaccharides) contents. Overall, the results indicate that no major changes in cell wall polysaccharide composition occurred during softening of ripening grape berries, but that significant modification of specific polysaccharide components were observed, together with large changes in protein composition.

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The composition of cell walls from grape skin in Vitis vinifera intraspecific hybrids

et al. 2017

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Isolation and characterization of cell walls from the mesocarp of mature grape berries (Vitis vinifera)

Cited by 23 publications

References 38 publications

Solubilization of an Arabinan Arabinosyltransferase Activity from Mung Bean Hypocotyls

Solubilization of an Arabinan Arabinosyltransferase Activity from Mung Bean Hypocotyls

Changes in Cell Wall Composition during Ripening of Grape Berries

The composition of cell walls from grape skin in Vitis vinifera intraspecific hybrids

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