Immunolocalization of pectin and hemicellulose epitopes in the phloem of Norway spruce and Scots pine

Distribution patterns of pectin and hemicellulose epitopes in the phloem of Pinaceae conifers differ between cell types including sieve cells, axial/ray parenchyma cells and stone cells, which is more prominent in seedlings than mature trees. Although there is considerable information on the gross chemistry of conifer bark, little is known on the chemistry of secondary phloem at the individual cell level. This study investigated distribution of pectins and hemicelluloses in the phloem of two conifer species (Norway spruce and Scots pine) at an individual cell wall level using nine monoclonal antibodies specific for pectin and hemicellulose epitopes combined with immunofluorescence and TEM immunogold labeling. Differences in phloem cell wall chemistry between juvenile (seedlings) and mature conifer trees were also examined. The two conifer species showed qualitatively similar distribution patterns of epitopes in sieve- and (axial/ray) parenchyma cells, irrespective of seedlings and mature trees. Sieve- and parenchyma cell walls showed the presence of rhamnogalacturonan-I (RG-I), homogalacturonan (HG) and xyloglucan epitopes, but revealed the absence of heteroxylan epitopes. Heteromannan epitopes were only detected in sieve cell walls, showing a chemical difference between sieve- and parenchyma cells. In contrast to qualitative similarity, there were several quantitative differences of epitope localization in sieve- and parenchyma cells between the two conifer species, indicating variations in the chemical structure and/or the amount of pectins and hemicelluloses between the two conifer species. These differences were more significant in seedlings than mature trees. Immunogold labeling of Norway spruce seedlings further indicated the possibility of chemical variations between cell wall regions within a single sieve cell wall. Phloem stone cells detected in mature Norway spruce showed the presence of heteromannans/heteroxylans and RG-I/HG/xyloglucans in secondary cell wall and middle lamellae, respectively.