Distributional variation of lignin and non-cellulosic polysaccharide epitopes in different pit membranes of Scots pine and Norway spruce seedlings

Abstract

Microdistribution of non-cellulosic polysaccharides in pit membranes of bordered pits (intertracheid pits between adjacent tracheids), cross-field pits (half bordered pits between tracheids and ray parenchyma cells) and ray pits (simple pits in nodular end walls of ray parenchyma cells) was investigated in mature earlywood of juvenile Scots pine and Norway spruce seedlings using immunocytochemistry combined with monoclonal antibodies specific to (1→4)-β-galactan (LM5), (1→5)-α-arabinan (LM6), homogalacturonan (HG, LM19, LM20), xyloglucan (LM15), xylan (LM10, LM11) and mannan (LM21, LM22) epitopes. Using phloroglucinol-HCl and KMnO4 staining, lignin distribution in pit membranes was also examined. Apart from cross-field pit membranes in Scots pine, all pit membranes observed showed a positive reaction for lignin with differences in staining intensity. Ray pit membranes showed strongest reaction with lignin staining in both species. Intensity of lignin staining in bordered pit membranes was stronger in Norway spruce than in Scots pine. With localization of non-cellulosic polysaccharide epitopes, Scots pine showed differences in cross-field pit membranes (rhamnogalacturonan-I (RG-I), HG and xyloglucan epitopes) from bordered and ray pit membranes (RG-I and HG epitopes). In contrast, Norway spruce showed significant differences in ray pit membranes (RG-I, HG, xyloglucan, xylan and mannan epitopes) from bordered and cross-field pit membranes (HG and no/trace amount of RG-I epitopes). Distributional differences in HG epitopes depending on antibody type/ membrane regions were also observed in cross-field pit membranes between the two species. Together, the results suggest that distribution patterns of lignin and non-cellulosic polysaccharides in pit membranes differ significantly between pit types and between Scots pine and Norway spruce. Compared with the same types of pit membranes in hardwoods, the results for Scots pine and Norway spruce (softwoods) differed significantly.