Abstract

Nascent pectin and glucuronoxylan were prepared from membrane-bound enzymes obtained from pea epicotyls. They had previously been shown to exhibit a protein- and pH-dependent pattern of binding to cell wall ghosts and to xy-loglucan extracted from cell walls prepared from pea epicotyls; maximum binding required a pH of 3-4, and the pres-ence of cell wall proteins, namely assemblins. To determine whether wall polysaccharides deposited in cell walls be-have in the same manner as nascent polymers, radioactively labeled EDTA-soluble polymers were prepared from newly-deposited pea epicotyl cell walls. Different enzyme treatments followed by column chromatography, in addition to complete acid hydrolysis followed by paper and thin layer chromatography, indicated the presence of pectin, to-gether with smaller amounts of glucuronoxylan, in this EDTA-soluble extract. These radioactively labeled polysaccha-rides were found to bind to cell wall ghosts and to xyloglucan extracted from the second and third internodes of pea epicotyls cell walls in a pH-dependent manner, similar to the binding pattern obtained with nascent polymers. Maxi-mum binding occurred at pH 3-4, and also required the presence of protein.

Highlights

  • The plant cell wall contains cellulose microfibrils embedded in a non-crystalline matrix

  • Uronic acids did not migrate under the experimental conditions used for the paper chromatography, so the acid hydrolysate was analyzed by Thin layer chromatography (TLC) using silica-coated plates

  • Nascent GAX was previously reported to bind to hemicellulose from pea epicotyls in a pH-dependent manner, with the highest binding at pH 3.5 - 4.0; the binding was thought to occur via non-covalent bonds [4]

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Summary

Introduction

The plant cell wall contains cellulose microfibrils embedded in a non-crystalline matrix. Nascent EDTA-soluble 14C-polysaccharides prepared from pea seedlings (Pisum sativum) consist of radioactively labeled pectins and glucuronarabinoxylans (GAX), with a significantly greater abundance of pectins [4]. These polysaccharides bound to xyloglucan in a pH-dependant manner, with the highest binding occurring at pH 3 - 4, which corresponds to the pH of a growing wall. The binding decreased to almost zero at pH 6 [5] This binding pattern indicates a significant role in cell-wall assembly during deposition and cell wall extension during growth. The present study aims to determine whether newly-deposited polysaccharides extracted from preformed cell walls behave in the same manner as nascent polymers, with respect to their interaction with cell-wall ghosts and with xyloglucan

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