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Microfibril angle (MFA) is perhaps the easiest ultrastructural variable to measure for wood cell walls, and certainly the only such variable that has been measured on a large scale. Because cellulose is crystalline, the MFA of the S2 layer can be measured by X-ray diffraction. Automated X-ray scanning devices such as SilviScan have produced large datasets for a range of timber species using increment core samples. In conifers, microfibril angles are large in the juvenile wood and small in the mature wood. MFA is larger at the base of the tree for a given ring number from the pith, and decreases with height, increasing slightly at the top tree. In hardwoods, similar patterns occur, but with much less variation and much smaller microfibril angles in juvenile wood. MFA has significant heritability, but is also influenced by environmental factors as shown by its increased values in compression wood, decreased values in tension wood and, often, increased values following nutrient or water supplementation. Adjacent individual tracheids can show moderate differences in MFA that may be related to tracheid length, but not to lumen diameter or cell wall thickness. While there has been strong interest in the MFA of the S2 layer, which dominates the axial stiffness properties of tracheids and fibres, there has been little attention given to the microfibril angles of S1 and S3 layers, which may influence collapse resistance and other lateral properties. Such investigations have been limited by the much greater difficulty of measuring angles for these wall layers. MFA, in combination with basic density, shows a strong relationship to longitudinal modulus of elasticity, and to longitudinal shrinkage, which are the main reasons for interest in this cell wall property in conifers. In hardwoods, MFA is of more interest in relation to growth stress and shrinkage behaviour.

Plants contain abundant autofluorescent molecules that can be used for biochemical, physiological, or imaging studies. The two most studied molecules are chlorophyll (orange/red fluorescence) and lignin (blue/green fluorescence). Chlorophyll fluorescence is used to measure the physiological state of plants using handheld devices that can measure photosynthesis, linear electron flux, and CO2 assimilation by directly scanning leaves, or by using reconnaissance imaging from a drone, an aircraft or a satellite. Lignin fluorescence can be used in imaging studies of wood for phenotyping of genetic variants in order to evaluate reaction wood formation, assess chemical modification of wood, and study fundamental cell wall properties using Förster Resonant Energy Transfer (FRET) and other methods. Many other fluorescent molecules have been characterized both within the protoplast and as components of cell walls. Such molecules have fluorescence emissions across the visible spectrum and can potentially be differentiated by spectral imaging or by evaluating their response to change in pH (ferulates) or chemicals such as Naturstoff reagent (flavonoids). Induced autofluorescence using glutaraldehyde fixation has been used to enable imaging of proteins/organelles in the cell protoplast and to allow fluorescence imaging of fungal mycelium.

SUMMARYHow the diverse polysaccharides present in plant cell walls are assembled and interlinked into functional composites is not known in detail. Here, using two novel monoclonal antibodies and a carbohydrate-binding module directed against the mannan group of hemicellulose cell wall polysaccharides, we show that molecular recognition of mannan polysaccharides present in intact cell walls is severely restricted. In secondary cell walls, mannan esterification can prevent probe recognition of epitopes/ligands, and detection of mannans in primary cell walls can be effectively blocked by the presence of pectic homogalacturonan. Masking by pectic homogalacturonan is shown to be a widespread phenomenon in parenchyma systems, and masked mannan was found to be a feature of cell wall regions at pit fields. Direct fluorescence imaging using a mannan-specific carbohydrate-binding module and sequential enzyme treatments with an endo-b-mannanase confirmed the presence of cryptic epitopes and that the masking of primary cell wall mannan by pectin is a potential mechanism for controlling cell wall micro-environments.

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