Tissue localization of phenolic compounds in plants by confocal laser scanning microscopy

Phenolic compounds are involved in many interactions of plants with their biotic and abiotic environment. These substances accumulate in different plant tissues and cells during ontogenesis and under the influence of various environmental stimuli, respectively. Studies on the tissue localization of phenolic compounds provide a fundamental prerequisite for understanding the ecological functions of these compounds. The present work shows the localization of various phenolics in cell walls, vacuoles, and associated with cell nuclei, in leaves of a monocotyledonous and a dicotyledonous plant, in a gymnosperm as well as in rhizomes of a horsetail by confocal laser scanning microscopy (CLSM). Using fresh plant material, it compares in detail the tissue localization of autofluorescent styrylpyrones and hydroxycinnamic acids and the visualization of epidermal flavonoid compounds using shift reagents like ammonia, and fluorescence-inducing reagents like Naturstoffreagenz A (diphenylboric acid 2-aminoethyl ester). The comparison of microscopic data obtained from different plant species shows the advantages and limitations of confocal laser scanning microscopy in ecological biochemistry of phenolic plant metabolites.