Abstract

Iodine staining has been used to study the orientation of cellulose microfibrils in wood using light microscopy. The aim of this work was to understand the exact nature of the staining reaction with iodine and to provide insight into the properties and organisation of the wood cell wall. Based on transmission electron microscopy it is apparent that precipitation of the iodine following treatment with nitric acid results in the formation of crystal cavities within the cell wall, which follow the orientation of the cellulose microfibrils. There is no evidence that iodine precipitates within “drying checks” as previously speculated. High resolution confocal reflectance microscopy of crystal cavity orientation indicates that the microfibril arrangement within pit borders can be both spiral and circular. Crystal cavities are much more abundant within the S1 layer than elsewhere. All of the cells examined had crystal cavities in the S1 region, which may be related to the reduced lignification at the S1/S2 boundary resulting in greater porosity of the cell wall at this location. Within the S2 region, clusters of crystal cavities are randomly distributed and occur in widely varying numbers among adjacent cell walls, suggesting variations in the porosity of the S2 wall within and among adjacent tracheids. Cavities form preferentially within more electron lucent regions of the cell wall. The random nature of crystal cavity formation within S2 clusters probably reflects the underlying random nature of the cell wall nanostructure. We conclude that iodine staining can provide important clues to the nanostructural properties of tracheid cell walls.