Pectin-like heteroxylans in the early-diverging charophyte Klebsormidium fluitans

The cell walls of charophytic algae both resemble and differ from those of land plants. Cell walls in early-diverging charophytes (e.g. Klebsormidiophyceae) are particularly distinctive in ways that might enable survival in environments that are incompatible with land-plant polymers. This study therefore investigates the structure of Klebsormidium polysaccharides.

The 'pectin' fraction (defined by extractability) of Klebsormidium fluitans, solubilized by various buffers from alcohol-insoluble residues, was digested with several treatments that (partly) hydrolyse land-plant cell-wall polysaccharides. Products were analysed by gel-permeation and thin-layer chromatography.

The Klebsormidium pectic fraction made up ~30-50 % of its alcohol-insoluble residue, was optimally solubilized at pH 3-4 at 100 °C, and contained residues of xylose ≈ galactose > rhamnose > arabinose, fucose, mannose and glucose. Uronic acids were undetectable, and the pectic fraction was more readily solubilized by formate than by oxalate, suggesting a lack of chelation. Some land-plant-targeting hydrolases degraded the Klebsormidium pectic fraction: digestion by α-l-arabinanase, endo-β-(1→4)-d-xylanase and α-d-galactosidase suggests the presence of β-(1→4)-xylan with terminal α-l-arabinose, α-d-galactose and (unexpectedly) rhamnose. 'Driselase' released oligosaccharides of xylose and rhamnose (~1:1), and graded acid hydrolysis of these oligosaccharides indicated a 'rhamnoxylan' with rhamnose side-chains. Partial acid hydrolysis of Klebsormidium pectic fraction released rhamnose plus numerous oligosaccharides, one of which comprised xylose and galactose (~1:2 Gal/Xyl), suggesting a galactoxylan. Lichenase was ineffective, as were endo-β-(1→4)-d-galactanase, endo-β-(1→4)-d-mannanase, β-d-xylosidase and β-d-galactosidase.

Klebsormidium pectic fraction possesses many land-plant-like linkages but is unusual in lacking uronic acid residues and in containing rhamnoxylan and galactoxylan domains. Uronic acids allow land-plant and late-diverging charophyte pectins to form Ca2+-bridges, facilitating cell-wall polymer association; their absence from Klebsormidium suggests that neutral heteroxylans rely on alternative cross-linking mechanisms. This lack of dependence on Ca2+-bridges might confer on Klebsormidium the ability to grow in the acidic, metal-rich environments that it tolerates.