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Plants 2015, 4(1), 29-43; doi:10.3390/plants4010029

Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay

School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New Zealand
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Academic Editor: Dilantha Fernando
Received: 29 October 2014 / Accepted: 6 January 2015 / Published: 14 January 2015
(This article belongs to the Special Issue Plant Cell Walls: Chemical and Metabolic Analysis)
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Abstract

Compression wood is a type of reaction wood formed on the underside of softwood stems when they are tilted from the vertical and on the underside of branches. Its quantification is still a matter of some scientific debate. We developed a new technique that has the potential to do this based on the higher proportions of (1→4)-β-d-galactans that occur in tracheid cell walls of compression wood. Wood was milled, partially delignified, and the non-cellulosic polysaccharides, including the (1→4)-β-d-galactans, extracted with 6 M sodium hydroxide. After neutralizing, the solution was serially diluted, and the (1→4)-β-d-galactans determined by an immuno-dot assay using the monoclonal antibody LM5, which specifically recognizes this polysaccharide. Spots were quantified using a dilution series of a commercially available (1→4)-β-d-galactan from lupin seeds. Using this method, compression and opposite woods from radiata pine (Pinus radiata) were easily distinguished based on the amounts of (1→4)-β-d-galactans extracted. The non-cellulosic polysaccharides in the milled wood samples were also hydrolysed using 2 M trifluoroacetic acid followed by the separation and quantification of the released neutral monosaccharides by high performance anion exchange chromatography. This confirmed that the compression woods contained higher proportions of galactose-containing polysaccharides than the opposite woods. View Full-Text
Keywords: Pinus radiata; coniferous gymnosperms; compression wood; (1→4)-β-d-galactans; immuno-dot assay; LM5 monoclonal antibody; monosaccharide composition; delignification Pinus radiata; coniferous gymnosperms; compression wood; (1→4)-β-d-galactans; immuno-dot assay; LM5 monoclonal antibody; monosaccharide composition; delignification
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Chavan, R.R.; Fahey, L.M.; Harris, P.J. Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay. Plants 2015, 4, 29-43.

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