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Plant and Cell Physiology Advance Access originally published online on May 31, 2007
Plant and Cell Physiology 2007 48(7):984-999; doi:10.1093/pcp/pcm068
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

In situ, Chemical and Macromolecular Study of the Composition of Arabidopsis thaliana Seed Coat Mucilage

Audrey Macquet1, Marie-Christine Ralet2, Jocelyne Kronenberger1, Annie Marion-Poll1 and Helen M. North1,*

1Laboratoire de Biologie des Semences, UMR 204, INRA, INAPG, Institut Jean-Pierre Bourgin, INRA, F-78026 Versailles Cedex, France
2INRA, UR1268 Biopolymères Interactions Assemblages, F-44000 Nantes, France

*Corresponding author: E-mail, helen.north{at}versailles.inra.fr; Fax, +33-1-30-83-31-11.


  Abstract

A comprehensive analysis was carried out of the composition of seed coat mucilage from Arabidopsis thaliana using the Columbia-0 accession. Pectinaceous mucilage is released from myxospermous seeds upon imbibition, and in Arabidopsis consists of a water-soluble, outer layer and an adherent, inner layer. Analysis of monosaccharide composition in conjunction with digestion with pectolytic enzymes conclusively demonstrated that the principal pectic domain of both layers was rhamnogalacturonan I, and that in the outer layer this was unbranched. The macromolecular characteristics of the water-soluble mucilage indicated that the rhamnogalacturonan molecules in the outer layer were in a slightly expanded random-coil conformation. The inner, adherent layer remained attached to the seed, even after extraction with acid and alkali, suggesting that its integrity was maintained by covalent bonds. Confocal microscopy and monosaccharide composition analyses showed that the inner layer can be separated into two domains. The internal domain contained cellulose microfibrils, which could form a matrix with RGI and bind it to the seed. In effect, in the mum5-1 mutant where most of the inner and outer mucilage layers were water soluble, cellulose remained attached to the seed coat. Immunolabeling with anti-pectin antibodies indicated the presence of galactan and arabinan in the inner layer, with the latter only present in the non-cellulose-containing external domain. In addition, JIM5 and JIM7 antibodies labeled different domains of the inner layer, suggesting the presence of stretches of homogalacturonan with different levels of methyl esterification.

Keywords: Arabidopsis thaliana - Cellulose - Pectin - Rhamnogalacturonan I - Seed mucilage

Abbreviations: BSA, bovine serum albumin; GLC, gas–liquid chromatography; HG, homogalacturonan; HP-SEC, high-performance size-exclusion chromatography; mum, mucilage modified; MALLS, multiple angle laser light scattering; PBS, phosphate-buffered saline; RGI, rhamnogalacturonan I; RGII, rhamnogalacturonan II

(Received April 10, 2007; Accepted May 27, 2007)
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