Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay
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 h...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2015-01-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2223-7747/4/1/29 |
| _version_ | 1818957977900548096 |
|---|---|
| author | Ramesh R. Chavan Leona M. Fahey Philip J. Harris |
| author_facet | Ramesh R. Chavan Leona M. Fahey Philip J. Harris |
| author_sort | Ramesh R. Chavan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-20T11:18:26Z |
| format | Article |
| id | doaj.art-857e84800a6d4832922d335cd816fcc0 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-12-20T11:18:26Z |
| publishDate | 2015-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-857e84800a6d4832922d335cd816fcc02022-12-21T19:42:33ZengMDPI AGPlants2223-77472015-01-0141294310.3390/plants4010029plants4010029Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot AssayRamesh R. Chavan0Leona M. Fahey1Philip J. Harris2School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New ZealandSchool of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New ZealandSchool of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland 1142, New ZealandCompression 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.http://www.mdpi.com/2223-7747/4/1/29Pinus radiataconiferous gymnospermscompression wood(1→4)-β-d-galactansimmuno-dot assayLM5 monoclonal antibodymonosaccharide compositiondelignification |
| spellingShingle | Ramesh R. Chavan Leona M. Fahey Philip J. Harris Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay Plants Pinus radiata coniferous gymnosperms compression wood (1→4)-β-d-galactans immuno-dot assay LM5 monoclonal antibody monosaccharide composition delignification |
| title | Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay |
| title_full | Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay |
| title_fullStr | Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay |
| title_full_unstemmed | Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay |
| title_short | Quantification of (1→4)-β-d-Galactans in Compression Wood Using an Immuno-Dot Assay |
| title_sort | quantification of 1 4 β d galactans in compression wood using an immuno dot assay |
| topic | Pinus radiata coniferous gymnosperms compression wood (1→4)-β-d-galactans immuno-dot assay LM5 monoclonal antibody monosaccharide composition delignification |
| url | http://www.mdpi.com/2223-7747/4/1/29 |
| work_keys_str_mv | AT rameshrchavan quantificationof14bdgalactansincompressionwoodusinganimmunodotassay AT leonamfahey quantificationof14bdgalactansincompressionwoodusinganimmunodotassay AT philipjharris quantificationof14bdgalactansincompressionwoodusinganimmunodotassay |