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Visualizing Localizations and Movement of Anthracene in Kandelia candel (L.) Druce Leaves by Fluorescence Microscopy

Abstract Here, for the first time, fluorescence microscopy (FM) was coupled with leaf autofluorescence to visualize and track trace levels of anthracene (An) in living Kandelia candel (L.) Druce leaves, without destructive chemical extraction techniques. The experimental results revealed that, after tracking over 96 hours, An adsorbed onto the leaf surface was moved through the upper cuticle and through the stomata of the lower side into the internal leaf tissues. An was identifiable in six separate locations within the K. candel leaves: the first hypodermis, the second hypodermis, and the upper palisade tissue moving from the upper cuticle; the lower hypodermis and the lower palisade tissue moving from the stomata of the lower side; and the spongy mesophyll moving from both sides. Under the same exposure conditions, the impact of An adsorbed at the center of the upper side of the leaf surface was much less than that at the edges of the leaf surface. However, after 96 hours of exposure to An, the amount of An transferred into the inner leaf tissues at the center of the leaf was higher than at the edges, which might provide important new information concerning how An enters, is moved, and is distributed within the K. candel leaves. Furthermore, this FM technique provided us with a noninvasive tool for visualizing and tracking the movement and storage locations of PAHs within K. candel leaves based only on the autofluorescence of leaf tissues as revealed by fluorescence microscopy.