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Pre-formed xyloglucans and xylans increase in molecular weight in three distinct compartments of a maize cell-suspension culture

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Abstract

Cultured cells of maize (Zea mays L.) were pulse-labelled with l-[1-3H]arabinose (Ara) and then monitored for 7 days. The 3H-hemicelluloses present in three compartments (protoplasm, cell wall and culture medium) were size-fractionated and the fractions assayed for [3H]xyloglucans and [3H]xylans. Protoplasmic [3H]xylans and [3H]xyloglucans initially (15 min after [3H]Ara-feeding) had weight-average relative molecular masses (M w) ≈ 0.5×106 and 0.3×106, respectively, both rising to 2×106 by 30 min. Thus, newly formed hemicellulose molecules were joined to other polymers, or to each other, presumably within Golgi vesicles. New 3H-hemicelluloses very rapidly bound to the cell wall; however, after 1 day, some [3H]xyloglucan and [3H]xylan was sloughed from the wall into the medium. The wall-bound [3H]xyloglucans were present in the form of extremely large complexes, of M w>17×106, even as early as 15 min after [3H]Ara-feeding. This M w is >70-fold greater than that observed by similar methods in cultures of a dicotyledon (Rosa sp.). Thus, during wall-binding, newly secreted xyloglucans greatly increased in size, possibly by transglucosylation. Some modest degradation (trimming) of wall-bound [3H]xyloglucan occurred later. The earliest wall-bound [3H]xylan had M w≈2×106, similar to the protoplasmic [3H]xylan; this increased to ≈4×106 by 6 h. For the first 2 days after [3H]Ara-feeding, the soluble extracellular 3H-hemicelluloses present in the culture medium had M w≈1×106–2×106, comparable to the protoplasmic hemicelluloses. However, between 2 and 3 days after [3H]Ara-feeding, the M w of the soluble extracellular [3H]xylans increased abruptly to ≈10×106; the soluble extracellular [3H]xyloglucans underwent a similar but more gradual increase in M w. Maize 3H-hemicelluloses thus underwent increases in M w in three episodes: (i) intra-protoplasmically, (ii) during wall-binding (especially xyloglucans), and (iii) after sloughing into the medium. Possible mechanisms and roles of these increases are discussed.

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Abbreviations

Ara:

arabinose

GPC:

gel-permeation chromatography

K av :

elution volume on GPC relative to V 0 (K av=0) and V i (K av=1)

MLG:

mixed-linkage glucan

M r :

relative molecular mass

M T :

tritium-average relative molecular mass

M w :

weight-average relative molecular mass

PyAW:

pyridine/acetic acid/water (1:1:23, by vol.) containing 0.5% (w/v) chlorobutanol

TFA:

trifluoroacetic acid

V 0 :

void volume (centre of 40-MDa dextran peak) on GPC

V i :

totally included volume (centre of sucrose peak) on GPC

XET:

xyloglucan endotransglucosylase (activity)

XTH:

xyloglucan endotransglucosylase/hydrolase

Xyl:

xylose

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Acknowledgement

E.M.K. thanks the BBSRC for a studentship, during tenure of which this work was done.

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  1. The Edinburgh Cell Wall Group, Institute of Cell and Molecular Biology, The University of Edinburgh, Daniel Rutherford Building, The King's Buildings, Edinburgh, EH9 3JH, UK

    Ellen M. Kerr & Stephen C. Fry

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  1. Ellen M. Kerr
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Correspondence to Stephen C. Fry.

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Kerr, E.M., Fry, S.C. Pre-formed xyloglucans and xylans increase in molecular weight in three distinct compartments of a maize cell-suspension culture. Planta 217, 327–339 (2003). https://doi.org/10.1007/s00425-003-1027-2

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